泻火化瘀通窍法改善豚鼠耳蜗微循环障碍的实验研究
摘要
目的：突发性耳聋是耳鼻咽喉科常见病、多发病，属中医“暴聋”范畴，目前发病率呈明显上升趋势。祖国医学在长期的耳聋诊治临床实践中摸索和积累了丰富的经验，许多复方制剂具有对耳蜗微循环障碍多靶点调控的作用，情志致病和心理因素已成为耳鸣耳聋的发病中和转归的重要因素之一。现代医学研究表明活血化瘀、改善微循环治疗有一定的疗效，因此将祖国医学清肝泻火法与活血化瘀法有机的结合起来形成泻火化瘀通窍法可能获得更好的疗效，大量的临床研究已经证实了这一点，其代表方剂为泻火治聋冲剂，经过大量的临床观察，有很好的疗效。本实验旨在（1）泻火化瘀通窍法对豚鼠耳蜗微循环障碍的影响。（2）泻火化瘀通窍法对光化学诱导法所致豚鼠耳蜗微循环障碍耳蜗组织VEGF及bFGFmRNA表达的影响。（3）泻火化瘀通窍法对光化学诱导法所致豚鼠耳蜗微循环障碍耳蜗组织iNOSmRNA、 MMP-2mRNA表达的影响。（4）泻火化瘀通窍法对光化学诱导法所致豚鼠耳蜗微循环障碍耳蜗组织Bcl-2mRNA、BaxmRNA及其蛋白表达的影响。通过以上实验来研究根据中医“火”、“瘀”致病理论创立的泻火化瘀通窍法对耳蜗微循环障碍的改善调控及分子生物学作用机制。
材料与方法：
动物造模与分组：采用光化学诱导法造模，造模成功后，将豚鼠分成5组，即正常组、模型组、行气组、活血化瘀组、泻火化瘀通窍组，每组10只。
实验方法：正常组：予0.9%生理盐水4ml/只/天，每日分二次给药，共连续灌胃10天；模型组：术后12h，予0.9%生理盐水4ml/只/天灌胃，每日分二次给药，共连续灌胃10天；行气组：术后12h予行气中药4ml/只/天灌胃（含生药1g），每日分二次给药，共连续给药10天；活血化瘀组：术后12h，予活血化瘀药物4ml/只/天灌胃（含生药2g），每日分二次给药，共连续给药10天。泻火化瘀通窍组：术后12h，予泻火化瘀通窍药物4ml/只/天灌胃（含生药4g），每日分二次给药，共连续给药10天。
实验指标：（1）各组豚鼠分别于造模后、用药前及停药后1天检测ABR阈值；（2）利用体式解剖显微镜进行螺旋韧带和耳蜗基底膜铺片，对血管纹和毛细胞进行观察；（3）利用TBA法和可见光法对耳蜗组织MDA及CAT进行检测；（4）采用实时荧光定量PCR方法检测耳蜗组织VEGF及bFGFmRNA表达；（5））采用实时荧光定量PCR方法检测耳蜗组织iNOSmRNA、 MMP-2mRNA表达；（6）采用实时荧光定量PCR方法检测耳蜗组织Bcl-2及Bax mRNA表达；（7）采用Western blot蛋白免疫印迹法检测VEGF、Bcl-2及Bax蛋白的表达。统计学处理：采用SPSS11.5软件包进行统计分析，计量资料实验数据以平均值±标准差（x|-±S）表示，单因素方差分析（One-Way ANOVA）,组间比较采用最小显著差法（LSD）方法，以P﹤0.05，P﹤0.01作为显著性和极显著性差异的标准。
结果：
1、泻火化瘀通窍组治疗后ABR阈值明显低于活血化瘀组及行气组P﹤0.05，且活血化瘀组治疗后ABR阈值明显低于行气组P﹤0.05；各组与正常组及模型组相比较均有显著性差异P＜0.01。
2、正常组螺旋韧带血管纹显示基本正常；模型组可见耳蜗螺旋韧带血管纹血管内大量微血栓形成，血管粗细明显不均，部分血管中断；行气组可见耳蜗螺旋韧带血管纹血管内大量微血栓形成，血管粗细明显不均，部分血管中断；活血化瘀组可见耳蜗螺旋韧带血管纹血管内微血栓，未见血管明显中断，较泻火化瘀通窍组微血栓多一些；泻火化瘀通窍组可见耳蜗螺旋韧带血管纹血管内少量微血栓，未见血管中断。
3、耳蜗基底膜铺片显微结构显示：正常组外毛细胞分布呈三排，显示清晰，排列整齐，分布均匀，无明显缺失；模型组见外毛细胞大部分变形，排列严重紊乱，崩解破坏，大片缺损，细胞连接消失，纤毛倒伏严重；行气组见外毛细胞部分变形，出现较多片状缺损，部分细胞连接消失；活血化瘀组见外毛细胞排列较为整齐，部分变形，部分出现坏死；泻火化瘀通窍组见外毛细胞排列整齐，散在缺失，个别轮廓不清。模型组及行气组外毛细胞损伤较重，泻火化瘀通窍组及活血化瘀组外毛细胞损伤相对较轻，且活血化瘀组损伤较泻火化瘀通窍组明显。
4、泻火化瘀通窍组MDA含量明显低于活血化瘀组及行气组P﹤0.05，而CAT含量明显高于活血化瘀组及行气组P﹤0.05；与正常组及模型组相比较均有极显著性差异P＜0.01。
5、各治疗组VEGFmRNA的表达均明显高于模型组p﹤0.01，且泻火化瘀通窍组高于活血化瘀组及行气组P﹤0.05，活血化瘀组高于行气组P﹤0.05。泻火化瘀组VEGFmRNA的表达量约是活血化瘀组的2.4倍，约是行气组的6.8倍，约是模型组的8.8倍；VEGF蛋白表达为行气组与模型组VEGF/β-actin相比较无显著性差异P＞0.05；活血化瘀组与行气组VEGF/β-actin相比较有显著性差异P﹤0.05；泻火化瘀组与活血化瘀通窍组VEGF/β-actin相比较有显著性差异P﹤0.05。
各治疗组bFGF mRNA的表达均明显高于模型组p﹤0.05，且泻火化瘀通窍组高于活血化瘀组及行气组p﹤0.05，活血化瘀组高于行气组p﹤0.05。泻火化瘀组bFGF mRNA的表达量约是活血化瘀组的1.4倍，约是行气组的4.2倍，约是模型组的6.3倍。
6、正常组与各治疗组iNOSmRNA的表达均明显低于模型组p﹤0.05，且泻火化瘀通窍组iNOSmRNA的表达量与活血化瘀组接近p﹥0.05，模型组iNOSmRNA的表达约是行气组的1.3倍，约是活血化瘀组的4.7倍，约是泻火化瘀通窍组的4.6倍；各治疗组MMP-2mRNA的表达均明显低于模型组P﹤0.05，模型组MMP-2mRNA的表达是是行气组的1.5倍；是活血化瘀组的2.4倍；是泻火化瘀通窍组的7.3倍。
7、各组Bax mRNA的表达均明显低于模型组P﹤0.05，且泻火化瘀通窍组低于活血化瘀组及行气组P﹤0.05，活血化瘀组低于行气组P﹤0.05。模型组Bax mRNA的表达量约是行气组的2.2倍；约是活血化瘀组的4.4倍；约是泻火化瘀通窍组的6.4倍；约是正常组的14.8倍。行气通组Bax mRNA的表达量约是活血化瘀组的2倍；约是泻火化瘀通窍组的3倍。活血化瘀组Bax mRNA的表达量约是泻火化瘀通窍组的1.5倍；各治疗组Bcl-2mRNA的表达均明显高于模型组P﹤0.05，且泻火化瘀通窍组高于活血化瘀组及行气通窍组P﹤0.05，活血化瘀组高于行气组P﹤0.05。泻火化瘀通窍组Bcl-2mRNA的表达量约是活血化瘀组的2.5倍，约是行气通窍组的6倍，约是模型组的17倍。
8、蛋白表达：Bcl-2/Bax比值泻火化瘀通窍组﹥活血化瘀组﹥行气组﹥模型组，且各组比较均有显著性差异,p﹤0.05。
结论：
1、泻火化瘀通窍法具有加强改善微循环及加强抗脂质过氧化的作用。
2、泻火化瘀通窍法可有效改善光化学法所致豚鼠耳蜗微循环障碍，且优于活血化瘀法
3、单纯行气法对光化学法所致豚鼠耳蜗微循环障碍无明显效果。
4、泻火化瘀通窍法能够明显提高豚鼠耳蜗微循环障碍耳蜗组织VEGF和bFGF的表达,优于活血化瘀法和行气法。
5、泻火化瘀通窍法可能通过促进VEGF和bFGF的表达，来促进新生血管的形成，改善微循环障碍。
6、泻火化瘀通窍法与活血化瘀法均能够抑制iNOS的表达，保护耳蜗组织的缺血损伤，明显优于行气法。
7、泻火化瘀通窍法、活血化瘀法与行气法均能够抑制MMP-2的表达。
8、Bcl-2和Bax基因和蛋白均参与豚鼠耳蜗微循环障碍损伤，而且损伤后Bcl-2/Bax比值下降是促进凋亡的重要因素。
9、泻火化瘀通窍法和活血化瘀法均能通过显著提高Bcl-2/Bax比值，抑制细胞凋亡。
10、泻火化瘀通窍法抑制细胞凋亡能力高于活血化瘀法。
11、 iNOS与Bc1-2的表达关系密切，iNOS的表达与Bcl-2/Bax比值呈负相关。
Objective：
Sudden deafness is a common disease in otolaryngology, attributed to "acutedeaf" category in Traditional Chinese Medicine (TCM). At present, it isincreasing in incidence. There in TCM clinical practice a rich experience hasbeen explored and accumulated through a long-term diagnosis and treatment ofdeafness. Many compound agents have the action of multi-targeted regulation ofcochlear microcirculation; and emotional and psychological factors have becomethe important factors in onset and conversion of deafness and tinnitus. Modernmedical research shows that activating blood to resolve the stasis, or improvingmicrocirculation therapy, has a definite effect. So a therapy of “reducing fireand resolving stasis to dredge the orifice”(RRDO) might have a better effect,which is derived from an organic integration of activating blood to resolve thestasis plus clearing away liver fire, and has been confirmed by a large numberof clinical studies. The representative prescription for the therapy is XiehuoZhilong Granules (Granules for Reducing Fire to Cure Deafness), which has a goodeffect in a large number of clinical observations. This study was designed forobserving the impacts of RRDO on:(1) disturbance of cochlear microcirculationin guinea pig;(2) VEGF and bFGFmRNA expression in cochlear microcirculationdisorder of guinea pigs induced by photochemical method;(3) expressions ofiNOSmRNA and MMP-2mRNA in cochlear microcirculation disorder of guinea pigsinduced by photochemical method;(4) expressions of BCL-2mRNA, BAX mRNA and theirprotein in cochlear microcirculation disorder of guinea pigs induced byphotochemical method. Through the above investigations we study the improvementand regulation of cochlear microcirculation, and molecular mechanism of RRDOestablished on the basis of pathogenesis by "fire" and "stasis".
Materials and methods:
Animal model and grouping: Photochemical induction method was used inmodeling, then the guinea pig models were divided into five groups, namely normalgroup, model group, qi-promoting group, blood-activating group, andfire-reducing group, with10guinea pigs in each group.
Experimental method:2ml of0.9%normal saline twice a day for the normalgroup;2ml of0.9%normal saline twice a day for the model group12-h after theoperation; and different treatments for the rest three groups. The specimensof the guinea pigs were sampled after quick decapitation on the2ndday afteradministration of anesthesia.(1) for fire-reducing group:12-h after theoperation, administration of the RRDO decoction,2ml, twice a day (amountingto1g/day for the raw medicine);(2) for the blood-activating group:administration of the ingredients activating blood to resolve stasis in the RRDOdecoction,2ml, twice a day (amounting to2g/day for the raw medicine);(3) forthe qi-promoting group: administration of the ingredients the qi-promotingsimply in the RRDO decoction,2ml, twice a day (amounting to4g/day for the rawmedicine). The course of treatment consisted of10days.
Experimental indicators:(1) ABR threshold detection was conducted for allthe guinea pigs after modeling, before and1day after medication;(2) the striavascularis and hair cells were observed by using microscope-body anatomy andcochlea spiral ligament and basilar membrane stretched;(3) detections of MDAand CAT were done with cochlear TBA method and Visible light method;(4)expressions of cochlear VEGFmRNA and bFGFmRNA were determined by using Real-TimePCR;(5)) cochlear iNOSmRNA, MMP-2mRNA expressions were detected with Real-TimePCR;(6) cochlear BAX mRNA and BCL-2mRNA expressions were determined withReal-Time PCR;(7) VEGF, BCL-2and BAX protein expressions were determined byusing western blot.
Statistical analysis: SPSS11.5package for statistical analysis was used.The measurement data of experimental data were expressed as (x±S); single-factor analysis of variance (One-Way ANOVA), and comparison betweengroups were conducted by using least significant difference method (LSD); andP﹤0.05, P﹤0.01were taken as significant and highly significant differencesin standards.
Results:
1. After the treatment, the ABR threshold was significantly lower in thefire-reducing group than in the blood-activating group and the qi-promotinggroup (P﹤0.05), and the ABR threshold was significantly lower in theblood-activating group than in the qi-promoting group (P﹤0.05). Compared withthe normal group and model group, the ABR thresholds in the three groups weresignificantly different (P﹤0.01).
2. In the normal group spiral ligament showed normal vascular stria; in themodel group a large number of micro-vascular thrombi could be seen in the vesselsof spiral ligament stria, with vascular thickening to different degree andvascular disruption of parts of the vessels; in the qi-promoting group a largenumber of micro-vascular thrombi could be seen in the vessels of spiral ligamentstria, with vascular thickening to different degree and vascular disruption ofparts of the vessels; in the activating blood group the micro-vascular thrombicould were observed in spiral ligament stria, but with no significant vasculardisruption, and more micro-vascular thrombi compared to the fire-reducing group;in the fire-reducing group only a small amount of micro-vascular thrombi couldbe seen in spiral ligament stria, with no vascular disruption.
3. The microstructure of the basilar membrane stretched: In the normal group,outer hair cells was distributed in three rows; clearly displayed, orderlyarranged, evenly distributed, and with no obvious missing; in the model group,most of the outer hair cells were deformed, arranged in serious disorder,disintegrated and destructed was lodged seriously, connections of celldisappeared, the cilia of cell was lodged seriously; in the qi-promoting group the part of outer hair cells was deformed, there were more flakes defect,connection of some cells disappeared; in the blood-activating group the outerhair cells arranged relatively orderly, partly deformed, and with partialnecrosis; in the fire-reducing group the outer hair cells arranged more orderly,missing sparsely, and with unclear contour occasionally. The damage of hair cellsin the model and the qi-promoting groups was serious, the damage of out haircell in the fire-reducing and the blood-activating groups the damage of outerhair cells was relatively mild, and the damage was more evident in theblood-activating group than in the fire-reducing group.
4. The MDA level of the fire-reducing group was significantly lower thanthose of the blood-activating and the qi-promoting groups (P﹤0.05), while CATlevel was significantly higher than those of the blood-activating and theqi-promoting groups (P﹤0.05). Compared with the normal group and model group,the three groups were significantly different in the two indicators(P﹤0.01).
5. The expressions of VEGFmRNA in all the3treatment groups weresignificantly higher than in the model group (p﹤0.01); it was higher in thefire-reducing group than in the blood-activating and the qi-promoting groups(P﹤0.05), and higher in the blood-activating group than in the qi-promotinggroup (P﹤0.05). The expression of VEGFmRNA in the fire-reducing group was about2.4times what it was in the blood-activating group, about6.8times what itwas in the qi-promoting group, and about8.8times what it was in the model group.For the expression of VEGF protein, in VEGF/β-actin there was no significantdifference between the qi-promoting group and the model group (P﹥0.05); therewas a significant difference between the blood-activating group and theqi-promoting group (P﹤0.05); and there was a significant difference betweenthe fire-reducing group and the blood-activating group (P﹤0.05). The expressionof bFGF mRNA of all the3treatment groups was significantly higher than thatof the model group (P﹤0.05); it was higher in the fire-reducing group than inthe blood-activating and the qi-promoting groups (P﹤0.05); and was higher inthe blood-activating group than in the qi-promoting group (P﹤0.05). The expression of bFGF mRNA in the fire-reducing group was about1.4times what itwas in the blood-activating group, about4.2times what it was in the qi-promotinggroup, and about6.3times what it was in the model group.
6. In the normal group and all the3treatment groups, the expressions ofiNOSmRNA were significantly lower than that in the model group (P﹤0.05). Theexpression of iNOSmRNA in the fire-reducing group was similar with that in theblood-activating (P﹥0.05). The expression of iNOSmRNA in the model group wasabout1.3times what it was in the qi-promoting group, about4.7times what itwas in the blood-activating group, and about4.6times what it was in thefire-reducing group. In every treatment group, the expression of MMP-2mRNA wassignificantly lower than that in the model group (P﹤0.05). In the model group,the expression of MMP-2mRNA was1.5times what it was in the qi-promoting group;2.4times what it was in the blood-activating group; and7.3times what it wasin the fire-reducing group.
7. In the normal group and all the3treatment groups, the expressions ofBAX mRNA were significantly lower than that in the model group (P﹤0.05). Theexpression of BAX mRNA in the fire-reducing group was lower than those in theblood-activating group and the qi-promoting group (P﹤0.05), the expression ofBAX mRNA in the blood-activating group was lower than that in the qi-promotinggroup (P﹤0.05). The expression of BAX mRNA in the model group was about2.2times what it was in the qi-promoting group o; about4.4times what it was inthe blood-activating group; about6.4times what it was in the fire-reducinggroup; and about14.8times what it was in the normal group. The expression ofBAX mRNA in the qi-promoting group was about2times what it was in theblood-activating group; about3times what it was in the fire-reducing group.The expression of BAX mRNA in the blood-activating group was about1.5timeswhat it was in the fire-reducing group. In the3treatment groups the expressionsof BCL-2mRNA were significantly higher than that the model group (P﹤0.05),and the expression of the fire-reducing group was higher than those in theblood-activating group and the qi-promoting group (P﹤0.05); the expression of the blood-activating group was higher than that in the qi-promoting group (P﹤0.05). In The fire-reducing group the expression of BCL-2mRNA was about2.5times what it was in the blood-activating group, about6times what it was inthe qi-promoting group, and about17times what it was in the model group.
8. In protein expression the ratio of BCL-2/Bax, the fire-reducing group﹥the blood-activating group﹥the qi-promoting group﹥the model group. Andthere was significant difference among all the groups (P﹤0.05).
Conclusion:
1. RRDO therapy has the functions of improving microcirculation andenhancing anti-lipid peroxidation.
2. RRDO therapy can effectively improve the photochemically induced cochlearmicrocirculation disturbance, and it is better than activating blood to resolvestasis.
3. The simple qi-promoting therapy has no evident effect on thephotochemically induced cochlear microcirculation disturbance
4.RRDO therapy can significantly improve the expression of VEGF and bFGFfor guinea pig with microcirculation disturbance of the cochlea, and the effectis better than those of activating blood to resolve stasis and promoting qi.
5. RRDO therapy may promote angiogenesis and improve microcirculationthrough promoting the expression of VEGF and bFGF.
6. Both RRDO therapy and activating blood to resolve stasis therapy caninhibit the expression of iNOS to protect cochlear tissue ischemia, and theyare better than qi-promoting therapy.
7. therapies of RRDO, activating blood to resolve stasis, and promoting qican all inhibit the expression of MMP-2.
8. Bcl-2and Bax genes and proteins are all involved in cochlearmicrocirculation damage. And the decreased Bcl-2/Bax ratio is an importantfactor in promoting apoptosis.
9. RRDO therapy and activating blood to resolve stasis therapy cansignificantly inhibit apoptosis through increasing the Bcl-2/Bax ratio.
10. In inhibiting apoptosis, RRDO therapy is better than activating bloodto resolve stasis therapy.
11. The expression of iNOS is closely related to Bc1-2, and iNOS expressionwas negatively correlated with the ratio of Bcl-2/Bax.
材料与方法：
动物造模与分组：采用光化学诱导法造模，造模成功后，将豚鼠分成5组，即正常组、模型组、行气组、活血化瘀组、泻火化瘀通窍组，每组10只。
实验方法：正常组：予0.9%生理盐水4ml/只/天，每日分二次给药，共连续灌胃10天；模型组：术后12h，予0.9%生理盐水4ml/只/天灌胃，每日分二次给药，共连续灌胃10天；行气组：术后12h予行气中药4ml/只/天灌胃（含生药1g），每日分二次给药，共连续给药10天；活血化瘀组：术后12h，予活血化瘀药物4ml/只/天灌胃（含生药2g），每日分二次给药，共连续给药10天。泻火化瘀通窍组：术后12h，予泻火化瘀通窍药物4ml/只/天灌胃（含生药4g），每日分二次给药，共连续给药10天。
实验指标：（1）各组豚鼠分别于造模后、用药前及停药后1天检测ABR阈值；（2）利用体式解剖显微镜进行螺旋韧带和耳蜗基底膜铺片，对血管纹和毛细胞进行观察；（3）利用TBA法和可见光法对耳蜗组织MDA及CAT进行检测；（4）采用实时荧光定量PCR方法检测耳蜗组织VEGF及bFGFmRNA表达；（5））采用实时荧光定量PCR方法检测耳蜗组织iNOSmRNA、 MMP-2mRNA表达；（6）采用实时荧光定量PCR方法检测耳蜗组织Bcl-2及Bax mRNA表达；（7）采用Western blot蛋白免疫印迹法检测VEGF、Bcl-2及Bax蛋白的表达。统计学处理：采用SPSS11.5软件包进行统计分析，计量资料实验数据以平均值±标准差（x|-±S）表示，单因素方差分析（One-Way ANOVA）,组间比较采用最小显著差法（LSD）方法，以P﹤0.05，P﹤0.01作为显著性和极显著性差异的标准。
结果：
1、泻火化瘀通窍组治疗后ABR阈值明显低于活血化瘀组及行气组P﹤0.05，且活血化瘀组治疗后ABR阈值明显低于行气组P﹤0.05；各组与正常组及模型组相比较均有显著性差异P＜0.01。
2、正常组螺旋韧带血管纹显示基本正常；模型组可见耳蜗螺旋韧带血管纹血管内大量微血栓形成，血管粗细明显不均，部分血管中断；行气组可见耳蜗螺旋韧带血管纹血管内大量微血栓形成，血管粗细明显不均，部分血管中断；活血化瘀组可见耳蜗螺旋韧带血管纹血管内微血栓，未见血管明显中断，较泻火化瘀通窍组微血栓多一些；泻火化瘀通窍组可见耳蜗螺旋韧带血管纹血管内少量微血栓，未见血管中断。
3、耳蜗基底膜铺片显微结构显示：正常组外毛细胞分布呈三排，显示清晰，排列整齐，分布均匀，无明显缺失；模型组见外毛细胞大部分变形，排列严重紊乱，崩解破坏，大片缺损，细胞连接消失，纤毛倒伏严重；行气组见外毛细胞部分变形，出现较多片状缺损，部分细胞连接消失；活血化瘀组见外毛细胞排列较为整齐，部分变形，部分出现坏死；泻火化瘀通窍组见外毛细胞排列整齐，散在缺失，个别轮廓不清。模型组及行气组外毛细胞损伤较重，泻火化瘀通窍组及活血化瘀组外毛细胞损伤相对较轻，且活血化瘀组损伤较泻火化瘀通窍组明显。
4、泻火化瘀通窍组MDA含量明显低于活血化瘀组及行气组P﹤0.05，而CAT含量明显高于活血化瘀组及行气组P﹤0.05；与正常组及模型组相比较均有极显著性差异P＜0.01。
5、各治疗组VEGFmRNA的表达均明显高于模型组p﹤0.01，且泻火化瘀通窍组高于活血化瘀组及行气组P﹤0.05，活血化瘀组高于行气组P﹤0.05。泻火化瘀组VEGFmRNA的表达量约是活血化瘀组的2.4倍，约是行气组的6.8倍，约是模型组的8.8倍；VEGF蛋白表达为行气组与模型组VEGF/β-actin相比较无显著性差异P＞0.05；活血化瘀组与行气组VEGF/β-actin相比较有显著性差异P﹤0.05；泻火化瘀组与活血化瘀通窍组VEGF/β-actin相比较有显著性差异P﹤0.05。
各治疗组bFGF mRNA的表达均明显高于模型组p﹤0.05，且泻火化瘀通窍组高于活血化瘀组及行气组p﹤0.05，活血化瘀组高于行气组p﹤0.05。泻火化瘀组bFGF mRNA的表达量约是活血化瘀组的1.4倍，约是行气组的4.2倍，约是模型组的6.3倍。
6、正常组与各治疗组iNOSmRNA的表达均明显低于模型组p﹤0.05，且泻火化瘀通窍组iNOSmRNA的表达量与活血化瘀组接近p﹥0.05，模型组iNOSmRNA的表达约是行气组的1.3倍，约是活血化瘀组的4.7倍，约是泻火化瘀通窍组的4.6倍；各治疗组MMP-2mRNA的表达均明显低于模型组P﹤0.05，模型组MMP-2mRNA的表达是是行气组的1.5倍；是活血化瘀组的2.4倍；是泻火化瘀通窍组的7.3倍。
7、各组Bax mRNA的表达均明显低于模型组P﹤0.05，且泻火化瘀通窍组低于活血化瘀组及行气组P﹤0.05，活血化瘀组低于行气组P﹤0.05。模型组Bax mRNA的表达量约是行气组的2.2倍；约是活血化瘀组的4.4倍；约是泻火化瘀通窍组的6.4倍；约是正常组的14.8倍。行气通组Bax mRNA的表达量约是活血化瘀组的2倍；约是泻火化瘀通窍组的3倍。活血化瘀组Bax mRNA的表达量约是泻火化瘀通窍组的1.5倍；各治疗组Bcl-2mRNA的表达均明显高于模型组P﹤0.05，且泻火化瘀通窍组高于活血化瘀组及行气通窍组P﹤0.05，活血化瘀组高于行气组P﹤0.05。泻火化瘀通窍组Bcl-2mRNA的表达量约是活血化瘀组的2.5倍，约是行气通窍组的6倍，约是模型组的17倍。
8、蛋白表达：Bcl-2/Bax比值泻火化瘀通窍组﹥活血化瘀组﹥行气组﹥模型组，且各组比较均有显著性差异,p﹤0.05。
结论：
1、泻火化瘀通窍法具有加强改善微循环及加强抗脂质过氧化的作用。
2、泻火化瘀通窍法可有效改善光化学法所致豚鼠耳蜗微循环障碍，且优于活血化瘀法
3、单纯行气法对光化学法所致豚鼠耳蜗微循环障碍无明显效果。
4、泻火化瘀通窍法能够明显提高豚鼠耳蜗微循环障碍耳蜗组织VEGF和bFGF的表达,优于活血化瘀法和行气法。
5、泻火化瘀通窍法可能通过促进VEGF和bFGF的表达，来促进新生血管的形成，改善微循环障碍。
6、泻火化瘀通窍法与活血化瘀法均能够抑制iNOS的表达，保护耳蜗组织的缺血损伤，明显优于行气法。
7、泻火化瘀通窍法、活血化瘀法与行气法均能够抑制MMP-2的表达。
8、Bcl-2和Bax基因和蛋白均参与豚鼠耳蜗微循环障碍损伤，而且损伤后Bcl-2/Bax比值下降是促进凋亡的重要因素。
9、泻火化瘀通窍法和活血化瘀法均能通过显著提高Bcl-2/Bax比值，抑制细胞凋亡。
10、泻火化瘀通窍法抑制细胞凋亡能力高于活血化瘀法。
11、 iNOS与Bc1-2的表达关系密切，iNOS的表达与Bcl-2/Bax比值呈负相关。
Objective：
Sudden deafness is a common disease in otolaryngology, attributed to "acutedeaf" category in Traditional Chinese Medicine (TCM). At present, it isincreasing in incidence. There in TCM clinical practice a rich experience hasbeen explored and accumulated through a long-term diagnosis and treatment ofdeafness. Many compound agents have the action of multi-targeted regulation ofcochlear microcirculation; and emotional and psychological factors have becomethe important factors in onset and conversion of deafness and tinnitus. Modernmedical research shows that activating blood to resolve the stasis, or improvingmicrocirculation therapy, has a definite effect. So a therapy of “reducing fireand resolving stasis to dredge the orifice”(RRDO) might have a better effect,which is derived from an organic integration of activating blood to resolve thestasis plus clearing away liver fire, and has been confirmed by a large numberof clinical studies. The representative prescription for the therapy is XiehuoZhilong Granules (Granules for Reducing Fire to Cure Deafness), which has a goodeffect in a large number of clinical observations. This study was designed forobserving the impacts of RRDO on:(1) disturbance of cochlear microcirculationin guinea pig;(2) VEGF and bFGFmRNA expression in cochlear microcirculationdisorder of guinea pigs induced by photochemical method;(3) expressions ofiNOSmRNA and MMP-2mRNA in cochlear microcirculation disorder of guinea pigsinduced by photochemical method;(4) expressions of BCL-2mRNA, BAX mRNA and theirprotein in cochlear microcirculation disorder of guinea pigs induced byphotochemical method. Through the above investigations we study the improvementand regulation of cochlear microcirculation, and molecular mechanism of RRDOestablished on the basis of pathogenesis by "fire" and "stasis".
Materials and methods:
Animal model and grouping: Photochemical induction method was used inmodeling, then the guinea pig models were divided into five groups, namely normalgroup, model group, qi-promoting group, blood-activating group, andfire-reducing group, with10guinea pigs in each group.
Experimental method:2ml of0.9%normal saline twice a day for the normalgroup;2ml of0.9%normal saline twice a day for the model group12-h after theoperation; and different treatments for the rest three groups. The specimensof the guinea pigs were sampled after quick decapitation on the2ndday afteradministration of anesthesia.(1) for fire-reducing group:12-h after theoperation, administration of the RRDO decoction,2ml, twice a day (amountingto1g/day for the raw medicine);(2) for the blood-activating group:administration of the ingredients activating blood to resolve stasis in the RRDOdecoction,2ml, twice a day (amounting to2g/day for the raw medicine);(3) forthe qi-promoting group: administration of the ingredients the qi-promotingsimply in the RRDO decoction,2ml, twice a day (amounting to4g/day for the rawmedicine). The course of treatment consisted of10days.
Experimental indicators:(1) ABR threshold detection was conducted for allthe guinea pigs after modeling, before and1day after medication;(2) the striavascularis and hair cells were observed by using microscope-body anatomy andcochlea spiral ligament and basilar membrane stretched;(3) detections of MDAand CAT were done with cochlear TBA method and Visible light method;(4)expressions of cochlear VEGFmRNA and bFGFmRNA were determined by using Real-TimePCR;(5)) cochlear iNOSmRNA, MMP-2mRNA expressions were detected with Real-TimePCR;(6) cochlear BAX mRNA and BCL-2mRNA expressions were determined withReal-Time PCR;(7) VEGF, BCL-2and BAX protein expressions were determined byusing western blot.
Statistical analysis: SPSS11.5package for statistical analysis was used.The measurement data of experimental data were expressed as (x±S); single-factor analysis of variance (One-Way ANOVA), and comparison betweengroups were conducted by using least significant difference method (LSD); andP﹤0.05, P﹤0.01were taken as significant and highly significant differencesin standards.
Results:
1. After the treatment, the ABR threshold was significantly lower in thefire-reducing group than in the blood-activating group and the qi-promotinggroup (P﹤0.05), and the ABR threshold was significantly lower in theblood-activating group than in the qi-promoting group (P﹤0.05). Compared withthe normal group and model group, the ABR thresholds in the three groups weresignificantly different (P﹤0.01).
2. In the normal group spiral ligament showed normal vascular stria; in themodel group a large number of micro-vascular thrombi could be seen in the vesselsof spiral ligament stria, with vascular thickening to different degree andvascular disruption of parts of the vessels; in the qi-promoting group a largenumber of micro-vascular thrombi could be seen in the vessels of spiral ligamentstria, with vascular thickening to different degree and vascular disruption ofparts of the vessels; in the activating blood group the micro-vascular thrombicould were observed in spiral ligament stria, but with no significant vasculardisruption, and more micro-vascular thrombi compared to the fire-reducing group;in the fire-reducing group only a small amount of micro-vascular thrombi couldbe seen in spiral ligament stria, with no vascular disruption.
3. The microstructure of the basilar membrane stretched: In the normal group,outer hair cells was distributed in three rows; clearly displayed, orderlyarranged, evenly distributed, and with no obvious missing; in the model group,most of the outer hair cells were deformed, arranged in serious disorder,disintegrated and destructed was lodged seriously, connections of celldisappeared, the cilia of cell was lodged seriously; in the qi-promoting group the part of outer hair cells was deformed, there were more flakes defect,connection of some cells disappeared; in the blood-activating group the outerhair cells arranged relatively orderly, partly deformed, and with partialnecrosis; in the fire-reducing group the outer hair cells arranged more orderly,missing sparsely, and with unclear contour occasionally. The damage of hair cellsin the model and the qi-promoting groups was serious, the damage of out haircell in the fire-reducing and the blood-activating groups the damage of outerhair cells was relatively mild, and the damage was more evident in theblood-activating group than in the fire-reducing group.
4. The MDA level of the fire-reducing group was significantly lower thanthose of the blood-activating and the qi-promoting groups (P﹤0.05), while CATlevel was significantly higher than those of the blood-activating and theqi-promoting groups (P﹤0.05). Compared with the normal group and model group,the three groups were significantly different in the two indicators(P﹤0.01).
5. The expressions of VEGFmRNA in all the3treatment groups weresignificantly higher than in the model group (p﹤0.01); it was higher in thefire-reducing group than in the blood-activating and the qi-promoting groups(P﹤0.05), and higher in the blood-activating group than in the qi-promotinggroup (P﹤0.05). The expression of VEGFmRNA in the fire-reducing group was about2.4times what it was in the blood-activating group, about6.8times what itwas in the qi-promoting group, and about8.8times what it was in the model group.For the expression of VEGF protein, in VEGF/β-actin there was no significantdifference between the qi-promoting group and the model group (P﹥0.05); therewas a significant difference between the blood-activating group and theqi-promoting group (P﹤0.05); and there was a significant difference betweenthe fire-reducing group and the blood-activating group (P﹤0.05). The expressionof bFGF mRNA of all the3treatment groups was significantly higher than thatof the model group (P﹤0.05); it was higher in the fire-reducing group than inthe blood-activating and the qi-promoting groups (P﹤0.05); and was higher inthe blood-activating group than in the qi-promoting group (P﹤0.05). The expression of bFGF mRNA in the fire-reducing group was about1.4times what itwas in the blood-activating group, about4.2times what it was in the qi-promotinggroup, and about6.3times what it was in the model group.
6. In the normal group and all the3treatment groups, the expressions ofiNOSmRNA were significantly lower than that in the model group (P﹤0.05). Theexpression of iNOSmRNA in the fire-reducing group was similar with that in theblood-activating (P﹥0.05). The expression of iNOSmRNA in the model group wasabout1.3times what it was in the qi-promoting group, about4.7times what itwas in the blood-activating group, and about4.6times what it was in thefire-reducing group. In every treatment group, the expression of MMP-2mRNA wassignificantly lower than that in the model group (P﹤0.05). In the model group,the expression of MMP-2mRNA was1.5times what it was in the qi-promoting group;2.4times what it was in the blood-activating group; and7.3times what it wasin the fire-reducing group.
7. In the normal group and all the3treatment groups, the expressions ofBAX mRNA were significantly lower than that in the model group (P﹤0.05). Theexpression of BAX mRNA in the fire-reducing group was lower than those in theblood-activating group and the qi-promoting group (P﹤0.05), the expression ofBAX mRNA in the blood-activating group was lower than that in the qi-promotinggroup (P﹤0.05). The expression of BAX mRNA in the model group was about2.2times what it was in the qi-promoting group o; about4.4times what it was inthe blood-activating group; about6.4times what it was in the fire-reducinggroup; and about14.8times what it was in the normal group. The expression ofBAX mRNA in the qi-promoting group was about2times what it was in theblood-activating group; about3times what it was in the fire-reducing group.The expression of BAX mRNA in the blood-activating group was about1.5timeswhat it was in the fire-reducing group. In the3treatment groups the expressionsof BCL-2mRNA were significantly higher than that the model group (P﹤0.05),and the expression of the fire-reducing group was higher than those in theblood-activating group and the qi-promoting group (P﹤0.05); the expression of the blood-activating group was higher than that in the qi-promoting group (P﹤0.05). In The fire-reducing group the expression of BCL-2mRNA was about2.5times what it was in the blood-activating group, about6times what it was inthe qi-promoting group, and about17times what it was in the model group.
8. In protein expression the ratio of BCL-2/Bax, the fire-reducing group﹥the blood-activating group﹥the qi-promoting group﹥the model group. Andthere was significant difference among all the groups (P﹤0.05).
Conclusion:
1. RRDO therapy has the functions of improving microcirculation andenhancing anti-lipid peroxidation.
2. RRDO therapy can effectively improve the photochemically induced cochlearmicrocirculation disturbance, and it is better than activating blood to resolvestasis.
3. The simple qi-promoting therapy has no evident effect on thephotochemically induced cochlear microcirculation disturbance
4.RRDO therapy can significantly improve the expression of VEGF and bFGFfor guinea pig with microcirculation disturbance of the cochlea, and the effectis better than those of activating blood to resolve stasis and promoting qi.
5. RRDO therapy may promote angiogenesis and improve microcirculationthrough promoting the expression of VEGF and bFGF.
6. Both RRDO therapy and activating blood to resolve stasis therapy caninhibit the expression of iNOS to protect cochlear tissue ischemia, and theyare better than qi-promoting therapy.
7. therapies of RRDO, activating blood to resolve stasis, and promoting qican all inhibit the expression of MMP-2.
8. Bcl-2and Bax genes and proteins are all involved in cochlearmicrocirculation damage. And the decreased Bcl-2/Bax ratio is an importantfactor in promoting apoptosis.
9. RRDO therapy and activating blood to resolve stasis therapy cansignificantly inhibit apoptosis through increasing the Bcl-2/Bax ratio.
10. In inhibiting apoptosis, RRDO therapy is better than activating bloodto resolve stasis therapy.
11. The expression of iNOS is closely related to Bc1-2, and iNOS expressionwas negatively correlated with the ratio of Bcl-2/Bax.
引文
[1]中华耳鼻咽喉头颈外科学分会.突发性聋的诊断和治疗[J].中华耳鼻咽喉头颈外科杂志，2006，41（5）:325.
[2]Olzowy B，Osterkorn D，Suckfull M，et al. The incidence of sudden hearingloss is greater than previously assumed[J]. MMW Fortschr Med,2005147(14):37-38.
[3]Chen YS, Emmerling O, Ilgner J. Idiopathic sudden sensorineural hearing lossin children[J]. International Journal of Pediatric Otorhinolaryngology，2005，69(6):817-821.
[4]马崧，史保轩.小儿突发性耳聋11例[J].实用儿科临床杂志，2002，17(4):385-386.
[5]Scasso CA，Bruschini L，Berrettini S，et al. Progressive sensorineural hearingloss from infectious agents[J]. Acta Otorhinolaryngol Ital.1998，18:51-54.
[6]丁怡冰.突发性耳聋研究新进展[J].职业与健康,2007，23（18）:1653-1655.
[7]Zientalska E， Moszynski B， Kapiszewska D， et al. The results of suddendeafness treatment[J]. Otolaryngol Pol，1998，52(5):707-712.
[8]Schweinfurth JM，Cacace AT. Cochlear ischemia induced by circulating ironparticles under magnetic control: an animal model for sudden hearing loss[J].Am J Otol，2000，21(5):636-640.
[9]Sauvaget E，Kici S， Petelle B， et al. Vertebrobasilar occlusive disorderspresenting as sudden sensorineural hearing loss[J]. Laryngoscope，2004，114(2):327-332.
[10]Kazunori T，Temyuki H，Yasuhiro K， et al. Bilateral deafness as a prodromalsymptom of basilar artery occlusion[J]. J Neurol Sci，2002，193(2):147-150.
[11]李爱华，吴意.突发性耳聋患儿血液流变学指标检测及分析[J].实用预防医学,2010，17(11):2296-2297.
[12]季念英.突发性耳聋的病因研究进展[J].济宁医学院学报，2006，29(3):87-89.
[13]刘铤.梅尼埃病和突发性聋诊治的若干问题[J].中华耳鼻咽喉科杂志，1997，32（2）:124-125.
[14]郑金秀，陈国春，庄金梅，等.36例突发性耳聋伴眩晕临床分析[J].福建医学杂志，2001，23(5):111-112.
[15]俸家富，李冰.突发性耳聋患者外周血T细胞亚群的变化[J]，江西医学检验，2003，21(1):3-4.
[16]Solares CA，Tuohy VK.ELISPOT determination of interferon gamma T-cellfrequencies in patients with autoimmune sensorineural hearing loss[J]. Methodsin Molecular Biology，2005，302(3):253-260.
[17]Cadoni G，Fetoni AR，Agostino S，et al. Autoimmunity in sudden sensorineuralhearing loss：possible role of anti-endothelial cell auto antibodies[J]. ActaOtolaryngol Suppl，2002，122(5):30-33.
[18]Hardin MF， Barker M， Neis PR. Sensorineural hearing loss as the resultof cliff jumping[J]. J Ark Med Soc，2003，99:327-330.
[19]Foster PK，Luebke AE. A model for perilymphatic fistula induced hearing lossin the guinea pig cochlea[J]. Hearing Research，2002，167(1):175-179.
[20]Ottaviani F, Cadoni G, Marinelli L,et al. Anti0-endothelial auto-antibodiesin patients with sudden hearing loss[J]. Laryngoscope，1999，109(7):1084-1087.
[21]Patscheke JH， Arndt J， Dietz K， et al. The prothrombin G20210A mutationis a risk factor for sudden hearing loss in young patients[J]. Thromb Haemost，2001，86:1118-1119.
[22]Rudack C，Langer C，Junker R. Platelet GPIaC807T polymorphism is associatedwith negative outcome of sudden hearing loss[J]. Hearing Research，2004，191(1):41-48.
[23]柴峰.王永钦教授辨证论治暴聋经验[J].中华实用中西医杂志，2003,16（11）:1636.
[24]林祖辉，叶杰清，徐璐洁.辨证分型治疗突发性耳聋56例观察[J].实用中医药杂志，2003,19（3）：117-119.
[25]赵俐，海波.中西医结合治疗突发性耳聋疗效观察[J].内蒙古中医药，2003，22（6）：19.
[26]戈言平，沈强，徐锦.突发性耳聋纯音测听、经颅多普勒检查与中医证型相关性研究[J].中国中医急症，2009,18（5）：748—750.
[27]葛灏.突发性耳聋听力损失程度与中医辨证分型关系初探.南通医学院学报[J]，1999，19（2）：231.
[28]Kanemaru S， Fukushima H， Nakamura H， et al. Alpha-interferon for thetreatment of idiopathic sudden sensorineural hearing loss[J].Eur ArchOtorhinolaryngol，1997，254(3):158-162.
[29]王铁，苏崇周,王尔琴等.突发性耳聋三种药物疗效分析[J]，中国中西医结合耳鼻咽喉科杂志1995，3（2）:5-7.
[30]刘志慧，杨静波，李玲.对突发性耳聋病因及治疗的评估[J].航空航天医药，1998，9(3):154-155.
[31]李德炳，冯勇，王槐富.突发性耳聋治疗的新进展[J].听力学及言语疾病杂志，2004，12(5):349-351.
[32]Dodson KM，Sismanis A. Intratympanic perfusion for the treatment of tinnitus[J].Otolaryngol Clin North Am，2004，37(5):991-1000.
[33]Hoffmann F,Beck C,Schutz A,et al. Ginkgo extract EGb761(tenobin)/HAESversus naftidrofury/(Dusodril)/HAES. A randomized study of therapy of suddendeafness[J]. Laryngorhinootologie,2001,121:579-584.
[34]胡红.川芎嗪治疗突发性耳聋72例报告[J].咸宁学院学报(医学版)，2006，20（2）：151-153.
[35]赵健林.葛根素治疗突发性耳聋30例疗效观察[J]，山西医药杂志，2010，（39）8：748-749.
[36]郭丽,赵玉林,李小萍，等.复方丹参注射液治疗突发性耳聋66例[J].中医研究，2006，（19）5：40-41.
[37]栗珂，李秀敏，黄铁强.脉络宁治疗突发性耳聋临床观察[J].黑龙江中医药，2002，（3）：24.
[38]Zivic L，Zivic D，Stojanovic S. Sudden hearing loss our experience intreatment with vasoactive therapy [J]. Srp Arh Celok Lek，2008，136(3):91-94.
[39]邓基波，孙奉乾.前列腺素E脂微球制剂治疗突发性耳聋的疗效观察[J].中日友好医院学报，2006,20（1）：38-39.
[40]张洪颖.长春西汀注射液治疗脑梗死疗效观察[J].中国误诊学杂志，2008，8(12):2859-2860.
[41]孙洁，吕义美.突发性耳聋的药物治疗[J].中国基层医药，2004，11(6):761.
[42]沈峰.长春西汀治疗突发性耳聋的疗效观察[J].中国全科医学，2010,11:1238.
[43]王卫国，许荣，姚茂军.突发性耳聋的早期治疗及超氧化物歧化酶和丙二醛检测[J].中华耳鼻咽喉科杂志，2002，37(4):274-276.
[44]骆文龙，刘世喜，梁传余，等.三磷酸胞苷加地塞米松等综合治疗突发性耳聋的临床观察[J].中华耳鼻咽喉科杂志，2002，37（4）:277-279.
[45]张仲华，师秀萍，桂训海.应用单唾液酸四己糖神经节苷脂联合巴曲酶治疗突发性耳聋的临床研究[J].承德医学院学报，2009,26（1）：28-30.
[46]徐蓓,邵健.甲钴胺的临床应用[J].现代中西医结合杂志，2006,15(7):972-973.
[47]杨长君.甲钴胺治疗突发性耳聋的疗效观察[J].中国实用神经疾病杂志，2009,12（1）：97.
[48]Stokroos RJ，Albers FW，Schitm J，et al. The etiology of Idiopathic suddensensorineural hearing loss： Experimental herpes-simplex virus infection ofthe inner ear[J]. Am J Otol，1998，(19)4:447-452.
[49]李和清，任基浩，卢永德.突聋血清病毒特异性IgM抗体及NO检测的初步研究[J].中国耳鼻咽喉颅底外科杂志，2002，8（2）：133-134.
[50]Berrocal JR，Ramirez-Camacho R. Sudden sensorineural hearing loss: supporting theimmunologic theory. The Annals of Otology, Rhinology, and Laryngology，2002，111(11):989-997.
[51]Scherer EQ，Herzog M，Wangemann P. Endothelin-1-induced vasospasms of spiralmodiolar artery are mediated by rho-kinase-induced Ca(2+) sensitization ofcontractile apparatus and reversed by calcitonin gene-relatedPeptide[J].Stroke，2002，33(12):2965-2971.
[52]蒋春荣，殷团芳，卢永德，等.突聋与微循环障碍的关系[J].中国耳鼻咽喉颅底外科杂志，1999，5（1）：51-53.
[53]Thomas J Haberkamp， Hasan M Tanyeri.Management of idiopathic-suddensensorineural hearing loss [J].The American Journal of Otology，1999，20(5):587-595.
[54]Cole RR，Jahrsdoerfer RA. Sudden hearing loss:an update[J]. The AmericanJournal of Otology，1988，9(3):211-215.
[55]Minoda R， Masuyama K， Habu K， et al. Initial steroid hormone dose in thetreatment of idiopathic sudden deafness [J]. American Journal of Otology，2000，21(6):819-825.
[56]Herr BD， Marzo SJ. Intratympanic steroid perfusion for refractory suddensensorineural hearing loss[J]. Otolaryngol Head Neck Surg，2005，132(4):527-531.
[57]周杰，罗仁瀚,黄云声.龙胆泻肝汤加减治疗突发性耳聋30例临床观察[J].长春中医药大学学报,2009,25(1):120-121.
[58]李可.李可老中医急危重症疑难病经验专辑[M].太原:山西科学技术出版社，2002:274-275.
[59]杜长河.补阳还五汤加减治疗突发性耳聋55例[J].实用中医药杂志,2007，23（10）：640.
[60]沈映群.中药药理学[M].上海：上海科学技术出版社，2000:136.
[61]朱天民，孙宏，金荣疆.活血化瘀方药对突发性耳聋患者BAEP及TCD脑血流参数的影响[J]，中国中西医结合杂志，2006,26（8）:740-742.
[62]郭伟，吴志学，闻留瑞.耳聋复聪汤为主治疗突发性耳聋30例临床观察[J].江苏中医药，2002,23(11):39.
[63]李彦华，邹广华.补肾活血法治疗突发性耳聋36例疗效观察[J].新中医，2008,40（12）：58-59.
[64]张弢.行气复聪方治疗突发性耳聋遗留耳鸣的THI评价[J].上海中医药杂志，2010，44(8):41-43.
[65]孔敏.龙胆泻肝汤加活血药治疗突发性耳聋临床观察[J].辽宁中医药大学学报，2009,11（2）：108-109.
[66]罗巧云，周和平，吕新莲,等.氦—氖激光血管内照射治疗Ⅱ型糖尿病临床和血液流变学的观察[J].中华理疗杂志,1997，20(2):95-98.
[67]郭桂荣，张淑萍，周志霞，等.低能量氦—氖激光血管内照射治疗高脂血症40例[J].中国疗养医学，1998，7(1):15-17.
[68]荆忱，戴成祥，刘瑞云，等.低能量氦—氖激光血管内照射治疗冠心病60例[J].中华理疗杂志,1997，20(2):89-91.
[69]魏万林，林毅勇，徐秋玲，等.低能量氦—氖激光血管内照射治疗动脉栓塞性剧痛一例[J].中华理疗杂志,1997，20(1):63.
[70]高平青，姚富通，李虹，等.氦—氖激光血管内照射并脉络宁治疗急性脑梗塞临床研究[J].中华理疗杂志,1996，19(2):99-101.
[71]谭其仁，姜乃明，夏军，等.低强度氦—氖激光兔耳血管内照射对微循环障碍的影响[J].中华理疗杂志,1996，19(3):141-143.
[72]韦兴昌，刘开祥，俸军林，等.低能量氦—氖激光血管内照射疗法[J].中华理疗杂志,1995，18(1):45-48.
[73]齐杰，张素杰，朴永芬.低能量He-Ne激光血管内照射治疗心脑血管病20例[J].中国疗养医学,1996，5(1):42-44.
[74]孙曰升，周勇，王桂兰.氦氖激光血管内照射综合治疗快速性心律失常60例观察[J].中华理疗杂志,1998，21(6):365-367.
[75]丁桂才，王祝庆.低能量He-Ne激光血管内照射治疗外伤后头痛26例[J].中国疗养医学,1998，7(2):30-31.
[76]樊凌冰.高压氧治疗突发性耳聋的临床观察[J].青海医药杂志，1997，27(1):32-33.
[77]谢鼎华，杨伟炎.耳聋的临床与基础[M].长沙:湖南科学技术出版社，2004:246-251.
[78]陈学华，宁文静，吴世安.100例突发性耳聋疗效分析[J].实用医技杂志，2008，15(5):658-659.
[79]徐小军，赵志刚，李燕.突发性耳聋的治疗方法进展[J].医药导报，2000，19（6）:584-586.
[80]兰志红，邹朝福.突发性耳聋的疗效探索[J].中国中西医结合耳鼻咽喉科杂志，2002，10（2）:58-60.
[81]罗思瑾.药物、高压氧、体外反搏治疗突发性耳聋的疗效分析[J].暨南大学学报(医学版)，1999，20(2):99-102.
[82]马向明，任莉贇.针刺为主治疗突发性耳聋28例临床观察[J].中国中医药科技,2010,17（1）:31.
[83]范新华，丁亚南，常向辉.针灸与药物治疗突发性耳聋疗效对比观察[J].中国针灸，2010，30（8）:630-632.
[84]谢强，任元元，李唯钢.转移兴奋灶针灸法为主治疗感音神经性耳鸣的临床观察[J].实用中西医结合临床，2009，9(3):15-21.
[85]罗仁瀚,周杰,黄云声,等.电针治疗突发性耳聋疗效对照观察[J].中国针灸,2009,29（3）:185-187.
[1]Umemura K，Kohno Y，matsuno H，et al．A mew model for photochemically inducedthrombosis in the inner ear microcirculation and the use of hearing loss as ameasure for mirocircularory disorders[J].Eur Arch Otorhinolaryngol,1990，248：105-108．
[2]Jin K,Yasuhiro I,Kaoru O,et al.Effect of single-drug treatment onidiopathicsudden sensorineural hearing loss[J].Auris Nasus Larynx,2003,30(2):123-127.
[3]张琳，马峰，方亚伦，等，高血压病中医辨证与甲襞微循环变化的关系[J].微循环学杂志，1995，5（1）：39.
[4]黄焱明，邓水平，何世银.高血压病中医辨证与红细胞变形性关系的研究[J].四川中医，1993，11（2）：11.
[5]郑峰，胡世云，孙光.Ⅱ期高血压病中医证型与血浆内皮素一氧化氮关系的临床研究[J].福建中医药，2000，31（2）：5.
[6]王玉林，董明敏.耳蜗微循环研究进展[J].微循环学杂志，2002，12（2）：35-38.
[7] Hirano K,Ikeda K, Kawase T. Prognosis of sudden deafness with specialreference to risk factors of microvascular pathology[J]. Auris NasusLarynx,1999,26(2):111-115.
[8] Suzuki A,Ren T,Nuttulall al,et al.Age-related changes in cochlear bloodflow response to occlusion of anterior inferior cerebellar artery in mice[J].AnnOtol Rhinol Laryngol,1998,107(8):648—653.
[9]Goldwin B, Khan MJ, Seidman MD, et al. Sarthran preserves cochlear microcirculation and reduces temporary threshold shifts after noiseexposure[J].Otolaryngol Head Neck Surg,1998,118(5):576—583.
[10]Nagure M, Iwasaki S,Wu R,et al. Effects of corticosterod, contrast mediumand ATP on focal microcirculatory disorders of the cochlea[J]. Eur JPharmacol,1999,366(1):47-53.
[11] Penha R，O’Neill MA, Goyri O’Neill J,et al.Ultratructural aspects of themicrovasculture of the cochlea:the internal spiral network[J].Otolaryngol HeadNeck Surg,1999,120:725-729.
[12] Aird WC. Endothelium as an organ system[J]. Crit Care Med,2004,32(5):271.
[13]林琳,孙建和,黄德亮,等.胎儿耳蜗血管纹的扫描电镜观察[J].临床耳鼻咽喉科杂志,2005,19:157.
[14] Nakasima T, Naganawa S,Sone M,et al.Disorders of cochlear bloodflow[J].Brain Reviews,2003,43:17.
[15] Ohlemiler KK,Wright JS,Dugan LL.Early elevation of cochlear reactive oxygenspecies following noise exposure[J].Audiol Neurootol,1999,4:229.
[16] Losano G,Gattullo D,Pagliaro P.Myocardial neural and vascular aspects ofischemic preconditioning[J].Life Sciences,1996,59:185.
[17] Yang Gy,Betz AL. Reperfusion-induced injury to the blood-brain barrierafter middle cerebral artery occlusion in rats[J].Stroke,1994,25（1）：658.
[18] Brechtel P,Nuttal,Miller JM. Basal nitric oxide production in regulationof cochlear blood flow[J]. Hearing Research,1994,77:38.
[19]叶放蕾,董明敏.豚鼠基底动脉缺血一再灌注耳蜗损伤的实验研究[J].郑州大学学报(医学版),2002,37:320.
[20] Opremcak EM，Bruce RA，Lomeo MD，et al．Radial optic neurotomy for centralretinal vein occlusion：a retrospective pilot study of11consecutivecases[J].Retina，2001，21：408-415.
[21]吴昊，周维镕．豚鼠耳蜗微循环障碍模型的实验研究[J].南通医学院学报，2003，23：246-248．
[22]胡博华，周娜．不同程度的阻断内耳供血对耳蜗微循环和耳蜗生理功能的影响[J].军医进修学院学报，1999，20：179-181．
[23]罗志强，孔维佳，张先锋，等．三氯化铁诱导建立基底动脉栓塞动物模型[J]．临床耳鼻咽喉科杂志，2003，17（6）：364-366．
[24]Takeshita T，Iwasaki S，Nagura M，et al．Photochemically induced double lateralwall lesions in the guinea pig cochlea[J]．Acta Otolaryngol，2003，123：355-361．
[25] Scheibe F，Haupt H，Grunert H．Laser Doppler measurements of inner ear bloodflow during experimental thrombosis of cochlear blood vessels in the guineapig[J].Eur Arch Otorhinolaryngol，1997，254：86-90．
[26]王玉林，董明敏.光化学诱导豚鼠耳蜗微循环障碍的实验研究[J]，中国微循环，2002，6（6）：338-340.
[27]何景春，周维镕，严之曼等.光化学法诱导豚鼠耳蜗微循环障碍模型，中国耳鼻咽喉头颈外科[J]，2007，14（6）：361-364.
[28]Kavak li A,Sahna E, Parlakpinar H, et al.The effects of melaton in on focalcerebral ischem iareperfusion model[J].Saudi Med J,2004,25(11):1751-1752.
[29]席恺,董明敏．α-硫辛酸对豚鼠缺血-再灌注耳蜗损伤的防治作用，郑州大学学报(医学版)[J]，2005，40（2）：268-270.
[30]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：1237.
[31]范金茹，等.参附注射液与柴胡注射液抗脂质过氧化的作用[J].中成药，1991，13（2）：25.
[32]太田康幸.柴胡皂苷增强糖皮质激素作用的机制[J].国外医学·中医中药分册，1981，3（3）：58.
[33]朱江，蔡德海，芮菁.栀子的抗炎镇痛作用研究[J].中草药，2000，（31）3：198.
[34]王本祥主编.现代中药药理学[M].第1版.天津：天津科学技术出版社，1997，1107.
[35]陈长勋，金若敏.钩藤碱对血小板聚集和血栓形成的抑制作用[J].中国药理学报，1992，13（2）：126-130.
[36]胡锦官，顾健，王志旺.石菖蒲及其有效成分对中枢神经系统作用的实验研究[J].中药药理与临床，1999，15（3）：19.
[37]李明亚，陈红梅.石菖蒲对行为绝望动物抑郁模型的抗抑郁作用[J].中药材，2001，24（1）：40.
[38]张信岳，郑高利，寿燕，等.石菖蒲的益智和抗惊厥作用研究[J].浙江中医学院学报,1999,23(2):46.
[39]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：416.
[40]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：1237.
[41]刘军，吴卫平，匡培根.丹参在缺血再灌注时的神经保护作用及成纤维细胞生长因子上调[J].中国神经免疫学和神经病学杂志，1996，3（4）：225.
[42]李菊，杨期东.微透析技术和丹参注射液对Wistar鼠脑缺血时海马细胞外液兴奋性氨基酸释放的影响[J].中风与神经疾病杂志，1995，12（5）：258.
[43]柯朝阳，吴展元，肖伯奎.丹参、辅酶Q10对低氧供豚鼠动脉血气及耳蜗电图的影响[J].实用儿科临床杂志，2000，15（3）：132.
[44]蔡庆，刘红巾.碱性成纤维细胞生长因子和丹参对反复高Gz暴露大鼠脑组织中Bcl-2和P53mRNA表达的影响.空军总医学院学报[J]，2000，16（3）：130.
[45]吴卫平，姜树军，张小澍. C-Jun、bFGF、和HSP70在丹参改善颞叶缺血大鼠空间记忆障碍中的变化[J].卒中与神经疾病，1999，6（4）：193-195.
[46]黄青青，杨镛，万林骏，等.复方丹参注射液对兔内毒素休克的防治作用[J].中国危重病急救医学,1998，10（4）：227.
[47]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：170.
[48]Li M, Handa S, Ikeda Y, et al.Specific inhibiting characteristics oftertramethylpyrazine,one of active ingredients of the Chinese herbal medicine‘Chuanxiong,’on platelet thrombus formation under high shear rates[J]. ThrombRes,2001,104(1):15-28.
[49]范立，吕爱刚.川芎嗪对离体兔血管平滑肌和豚鼠盲肠带的作用[J].新药与临床，1997，16（4）：211.
[50]曲友直，高国栋，赵燕玲.川芎嗪、黄芪合用对脑缺血再灌注后脑组织C-Jun蛋白表达影响[J].安徽中医学院学报，2000，19（6）：256.
[51]Shih YH, Wu SL, Chiou WF, et al. Protective effects oftetramethylpyrazine on kainite-induced excitotoxicity in hippocampal culture.Neuro report[J],2002,13(4):515-519.
[52]蔡晨，等.川芎嗪对烫伤豚鼠血管内皮细胞保护作用的实验研究[J].中国烧伤创疡杂志，2000，（4）：11.
[53]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：1504—1507.
[54]Zhang GC.Antioxidation of pueraria lobata isoflavones[J].CA,1999,130:105.
[55]洪俐.葛根的研究进展概述.基层中医药杂志[J]，2000，14（2）：52.
[56]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：657-661.
[57]金鸣，李金荣.红花水溶性成分抗氧化作用的研究[J].心肺血管病杂志，1998，17（4）：277.
[58]张明霞，赵惠.红花对小鼠抗应激能力的影响[J].中国中医药信息杂志，1999，6（1）：26.
[59]马亚兵.枳壳的胃肠作用及炮制前后的变化[J].中药药理与临床，1996，12（6）：28.
[60]明彩荣，刘海星，井欢.中药抗过敏反应的显微微分干涉显微镜观察[J].中医药学刊，2001，19（3）：275.
[61]杨俊业，熊天琦.夜交藤煎剂的镇静催眠作用[J].华西医科大学学报，1990，21（2）：175.
[62]黄树莲，陈学芬.首乌藤降脂作用的实验研究[J].中草药，1991，22（3）：117.
[63]薛建欣，严永清，蒋莹.黄芪、归尾、香附、川芎、赤芍等配伍对“血瘀”大鼠血液流变学的影响.中国中药杂志，1994,19（2）:108.
[64]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：428-429.
[65]包金凤，杨永健.甘草次酸钠对大鼠化学性腹膜炎的影响[J].中国药理学报，1997，18（3）：277.
[66]Kenzo Ohtsuki,Sanae Nakamura, Yoshihito Shimoyama, et al. A96-kDaglycyrrhizin-binding protein(gp96) from soybeans acts as a substrate for caseinkinase Ⅱ, and is highly related to lipoxygenase3[J]. The Journal ofBiochemistry,1995,118(6):1145-1150.
[67]Harada S, Karino A, Shimoyama Y,et al.Identification ofglycyrrhizin-binding protein kinase as casein Ⅱ and characterization of itsassociated phosphate acceptors in mouse liver[J].Biochem Biophys ResCommum,1996,227(1):102-109.
[68]句海松，忻文娟.甘草类黄酮对脂质过氧化和活性氧自由基的作用[J].药学学报，1998，24（11）：807.
[69]刘蕊、安杨.突发性耳聋证治的现代文献研究[J].新疆中医药，2009，27（3）：11-13.
[70]王高华，黄从新.青年抑郁障碍患者的血浆血小板因子4浓度[J].微循环学杂志，2003，13（2）：36-39.
[71]杨斌，叶兰仙，刘建斌，等.抑郁症患者血小板膜表面P-选择素的表达研究[J].中国神经精神疾病杂志，2005，31（1）：37-39.
[72]Delisi JS, Konopka JM, Russell K, et al. Platelet cytosolic calciumhyperresponsivity to serotonin in patients with hypertension and de-pressivesymptoms[J]. Biological Psychiatry,1999,45(8):1035-1041.
[73]钱欣梅,张向阳.氧自由基与耳聋[J].济宁医学院学报,2003，26(2):67.
[74]Coakun O, Ocakei A, Bayraktaroglu T, et al. Exercise training prevents andstreptozotocin-induced oxidative stress and β-cell damage in rat pancieas[J].Tohoku J Exp Med,2004,203(3):145-154.
[75]杨小莹，马世平，瞿融.柴胡提取物对慢性应激抑郁大鼠脂质过氧化及淋巴细胞增殖的影响[J].中国药科大学学报，2007，38（5）：442-445.
[76]Wichers M, Maes M. The psychoneuroimmuno-pathophysiology ofcytokine-induced depression in humans[J]. Int J Neuropsychopharmacol,2002,5(4):375-388.
[77]季宁东，李娟好，李明亚.石菖蒲提取液的抗抑郁作用及柴胡皂苷对其作用的影响[J].南京医科大学学报，2006，26（12）：1203-1206.
[78]王宝亮，苏谨.龙胆泻肝汤联合巴曲酶治疗突发性耳聋30例疗效观察[J].中国实用神经疾病杂志，2010，13（3）：79—80.
[79]Achen MG, Stacker SA.The vascular endothelial growth factor family proteinswhich guide the development of the vasculature[J]. Int J ExpPath,1998,79:255-265.
[80]李蕾，刘开祥. VEGF在缺血性脑血管病中研究进展[J].牡丹江医学院学报，2010，31（5）：69-71.
[81]Tuomisto TI, Rissanen TT, Vajanto I, et al.HIF-VEGF-VEGFR-2, TNF alpha andIGF pathways are up regulated in critical human skeletal muscle ischemia asstudied with DNA array[J].Atherosclerosis,2004,174:11-20.
[82]郭敬，王烈.血管内皮生长因子与治疗性血管生成研究进展[J].福州总医院学报，2010，17（1）：56-58.
[83]Shen F, FanY, SuH, et al. Adeno-associated viral vector-mediated hypoxia-regulated VEGF gene transfer promotes angiogenesis following focal cerebralischemia in mice[J]. Gene Ther,2008,15(1):30-39.
[84]Huang YF, Zhuang SQ, Chen DP, et al. Angiogenesis and its regulatory factorsin brain tissue of neonatal rat hypoxic-ischemic encephalopathy[J]. ZhonghuaErKe Za Zhi2004,42(3):210-214.
[85]梅爱农,姜亚平.缺血性脑血管病与血管生成[J].国外医学脑血管疾病分册,2004,12(1):67-70.
[86]Sengupta S, Toh SA, Sellers LA, et al. Modulating angiogenesis: the yin andthe yang in ginseng[J].Circulation,2004,110(10):1219.
[87]]王建湘,王行宽,尹树忠.心痛治肝法“心痛灵Ⅱ号”对冠心病心绞痛患者血脂代谢及内皮功能的影响[J].中国中医急症,2003,12(5):392.
[88]王文,王宗仁,张金洲,等.芪丹通脉片对大鼠缺血心肌的血管新生作用与机制研究[J].中国中医急症,2007,16(2):190.
[89]王振涛,韩丽华,朱明军,等.黄酮皂苷类中药对促心肌梗死后大鼠缺血心肌血管新生作用及相关生长因子表达的影响[J].中西医结合心脑血管病杂志,2006,4(4):305.
[90]]张喜成,何延政,李跃武,等.补肾生血胶囊动员骨髓内皮祖细胞促进缺血肢体血管新生的研究[J].江苏中医药,2006,27(3):57
[91]Yang ZJ, Bao WL, Qiu MH, et al. Role of vascular endothelial growth factorin neuronal DNA damage and repair in rat brain follow-ing a transient cerebralischemia［J. J Neurosci Res,2002,70(2):140-149.
[92]Bottcher R T，Niehrs C．Fibroblast growth factor signaling during earlyvertebratedevelopment［J］．Endocrine Rev，2005，26（1）：63－77．
[93］Damico F M．Angiogenesis and retinal diseases［J］．Arq Bras Oftlmol，2007，70（3）：547－553．
[94]王晶，尹丽娅，李庚山，等．bFGF对缺氧条件下培养的人脐静脉内皮细胞增殖影响［J］．武汉大学学报：医学版，2002，23（4）：298－300．
[95]Hall H．Modified fibrin hydrogel matrices：both，3D-scaffolds and local andcontrolled release systems to stimulate angiogenesis［J］．Curr Pharm Des，2007，13（35）：3597－3607．
[96]Koivisto H，Hyvarinen M，Stromberg AM，et al．Cultures of human embryonicstem cells：serum replacement medium or serum-containing media and the effectof basic fibroblast growth factor［J．Reprod Biomed Online，2004，9（3）：330－337．
[97]Cutroneo K R．Gene therapy for tissue regeneration［J］．Cell Biochem，2003，88（2）：418－425．
[98]Coumoul X，Deng C X．Roles of FGF receptors in mammalian development andcongenital diseases［J］．Birth Defects Res C Embryo Today，2003，69（4）：286－304．
[99]Mueller KL, Jacques BE, Kelley MW. Fibroblast growth factor signalingregulates pillar cell development in the organ of Corti[J]. J Neurosc,2002,22(21):9368-9377.
[100]Carmeliet P. Mechanisms of angiogenesis and arteriogenesis［J］. NatureMedicine,2000,6(4):389-395.
[101]郑鸣，王丰，梁书锋.豚鼠耳蜗缺血再灌注损伤后iNOS活性与细胞凋亡关系[J].中国临床解剖学杂志，2006.24（2）：190-193.
[102] Ruan RS, Leong SK, YeohKH. Effects of nitric oxide on normal and ischemiccochlea of the guinea pig[J]. Exp Neuro,l2001,169(1):200-207.
[103]胡安，金晓杰.耳蜗微循环缺血与再灌注损伤的研究进展[J].听力学及言语疾病杂志，2008，1（5）：437-438.
[104]熊巍，邱建华.耳蜗的缺血/再灌注损伤研究进展[J].国外医学耳鼻咽喉科学分册，2005,29（4）：201-203.
[105]Cash D,Beech JS,Rayne RC,et al. Neuroprotective effect of aminoguanidineon transient focal ischaemia in the rat brain[J].Brain Res,2001,29(1):91-103.
[106]叶放蕾,董明敏.豚鼠基底动脉缺血一再灌注耳蜗损伤的实研究[J].郑州大学学报(医学版),2002,37:320.
[107]Lepore DA.Nitric oxide synthase-independent generation of nitric oxide inmuscle ischemia-reperfusiion injury[J].Nitric Oxide，2000,4(6):541-545.
[108]Seidman MD,Bai U,Khan MJ,et al.Mitochondrial DAN deletion associated withaging and presbyacusis[J].Arch Otolaryngol Head Neck Surg，1997,123(10):1039-1045.
[109]Berges A,Van Nassauw L,Bosmans J,et al.Role of nitric oxide and oxidativestress in ischemic myocardial injury and preconditioning[J]. ActaCardiol,2003,58(2):119-132.
[110]邓芬,胡长林,谢运兰.丹红注射液对大鼠局灶脑缺血后一氧化氮合酶的影响[J].中西医结合心脑血管病杂志，2006，4（10）：880-881.
[111]Moncada S,Palmer RM. Nitric oxide: physiology,pathophysiology andpharmacology[J]. Pharmacological Reviews,1991,43(2):109-142.
[112]Danielisova V, Nemethova M, Burda J.The protective effect ofaminoguanidineon cerebral ischemic damage in the rat brain[J]. Physiol Res,2004,53(5):533-540.
[113]陈山泉.血府逐瘀汤治疗冠心病心绞痛的临床研究[J].中医函授通讯,1999,18(6):48.
[114]洁西卡，桃李，章李军,等.诱导型一氧化氮合酶抑制剂研发现状[J].现代生物医学进展，2007，17（11）：1741-1743.
[115]Chambers AF，Matrisian LM．Changing views of the role of matrixmetalloproteinases metalloproteinases in metastasis［J］.J Natl Cancer Inst，1997，89(17):1260-1270．
[116]姜双，陈晓隆. ROP小鼠视网膜组织中MMP-2、VEGF的表达变化及意义[J].山东医药，2010，50（42）：28-29.
[117]Turpeenniemi-Hujanen T.Gelatinases(MMP-2and-9)and their naturalinhibitors as prognostic indicators in solid cancers[J].Biochimie,2005,87(3):287-297.
[118]Nagel S, Su Y, Horstmann S,et al. Minocycline and hypothermia forreperfusion injury after focal cerebral ischemia in the rat: effects on BBBbreakdown and MMP expression in the acute and subacute phase [J]. Clin Chem,2010,56:21-33.
[119] Pfefferkorn T,Rosenberg GA.Closure of the blood-brain barrier by matrixmetalloproteinase inhibition reduces rtPA-mediated mortality in ce-rebralischemia with delayed reperfusion[J].Stroke,2003,34(8):2025-2030.
[120]张会超，韩丽华，王振涛，等.益气活血方对心梗后左室重构大鼠心肌基质金属蛋白酶-2及转化生长因子β1的影响[J].中成药，2010,32（11）：1871-1874.
[121]伍尚敏，张蔓菁，刘宾,等.地塞米松对血管瘤内皮细胞分泌MMP-2的影响[J].中国美容医学，2010，19（11）：1645-1649.
[122]陈彩萍,过晶.心通口服液对冠心病心绞痛患者血MMP-2、MMP-9的影响[J].实用临床医药杂志，2010,14（21）：87-89.
[123]雷燕，杨静，赵浩，等.人参三七川芎提取物延缓衰老小鼠血管老化的实验研究[J].中国中西医结合杂志，2010,30（9）：946-951.
[124]] Harkness KA, Adamson P, Sussman JD, et al. Dexamethasone regulation ofMatrix metalloproteinase expression in CNS vascular endothelium [J]. Brain,2000,123(4):698-709.
[125]杨爱东,李文雯,汪东颖，等.清热燥湿方对急性肺损伤大鼠肺组织基质金属蛋白酶2及其抑制因子基因表达的影响[J].中国中医药信息杂志,2010,17(8):27-29.
[126] Jialong Guo,Kailun Zhang,Yanmei Ji, et al. Effects of ethyl pyruvate onmyocardial apoptosis and expression of Bcl-2and Bax proteins after ischemiareperfusion in rats［J］.Huazhong Univ Sci Technol Med Sci,2008,28(3):281-3.
[127]Li JS, Zhang W,Kang ZM, et al. Hyperbaric oxygen preconditioningreduces ischemia-reperfusion injury by inhibition of apoptosis viamitochondrial pathway in rat brain［J］. Neuroscience.2009,159(4):1309-1315
[128] Cardoso CM, Almeida LM, Custodio JB. Protection of tamoxifen againstoxidation of mitochondrial thiols and NAD(P)H underlying the permeabilitytransition induced by prooxidants[J]. Chem Biol Interact,2004,148:149-161.
[129]Lawrence MS, Mclaughlin JR, Sun GH, et al. Herpes simplex viral vectorsexpressing Bcl-2are neuroprotective when delivered after a stroke[J].J CerebBlood FlowMetab,1997,17:740-744.
[130] Antonawich FJ,Federoff HJ,Davis IN.Bcl-2trannsduction, using a herpessimplex virus amplicon, protects hippocampal neurons from transient globalischemia[J].Exp Neurol,1999,156:130-137.
[131] Jayasurya A,Dheen ST,Yap WM,et al. Inducible nitric oxide syn-thase andBc1-2expression in nasopharyngeal cancer: correlationwith outcome ofpatientsafter radiotherapy[J]. Int JRadiatOncol BiolPhys,2003,56(3):837-845.
[132] Shimizu S，Nagayama T，Jin KL，et al.Bcl-2antisense treatment preventinduction of tolerance to focal ischemia in the rat brain［J］.Cereb Blood FlowMetab，2001，21(3):233-243.
[133]李浩,刘开祥.丹参酮ⅡA对鼠脑缺血再灌注损伤Bcl-2和caspase-3表达的影响[J].中国康复医学杂志,2008,23(8):691-693
[134]戴雍月,徐正衸,王万铁等.红花注射液对肺缺血再灌注损伤时细胞凋亡及Bcl-2和Bax基因的影响[J].中国动脉硬化杂志，2007,15（6）：415-419.
[135]许益笑，王万铁，徐正衸.葛根素对兔肺缺血再灌注损伤时Bcl-2、Bax及caspase-3的影响[J].温州医学院学报,2007,37（2）：126-129.
[136]胡永红，周爱民，田野，等.川芎嗪促进脑缺血耐受形成的作用及机制研究[J].中国中医基础医学杂志，2010,16（8）：671-674.
[137]樊蔚虹,姚建平,赵文景.柴胡疏肝散对卒中后抑郁大鼠海马组织Bcl-2,Bax蛋白表达的影响[J].中国实验方剂学杂志，2011.11（3）：181-183.
[138]Peng CH, TangTH,Huang CN, et al. Optosis induced by penta-acetyl geniposidein C6glioma cells is associated with JNK activation and Fas ligand induction[J].Toxicol Appl Pharmacol,2005,202(2):172-179.
[139]方永奇,匡忠生,谢宇辉,等.石菖蒲对缺血再灌注脑损伤大鼠神经细胞凋亡的影响[J].现代中西医结合杂志，2002，11(17):1647-1649.
[140]蒋建刚，吴基良，陈金和.甘草酸二铵对大鼠心肌缺血再灌注损伤后细胞凋亡的影响[J].微循环学杂志，2004,14（4）：31-33.
[2]Olzowy B，Osterkorn D，Suckfull M，et al. The incidence of sudden hearingloss is greater than previously assumed[J]. MMW Fortschr Med,2005147(14):37-38.
[3]Chen YS, Emmerling O, Ilgner J. Idiopathic sudden sensorineural hearing lossin children[J]. International Journal of Pediatric Otorhinolaryngology，2005，69(6):817-821.
[4]马崧，史保轩.小儿突发性耳聋11例[J].实用儿科临床杂志，2002，17(4):385-386.
[5]Scasso CA，Bruschini L，Berrettini S，et al. Progressive sensorineural hearingloss from infectious agents[J]. Acta Otorhinolaryngol Ital.1998，18:51-54.
[6]丁怡冰.突发性耳聋研究新进展[J].职业与健康,2007，23（18）:1653-1655.
[7]Zientalska E， Moszynski B， Kapiszewska D， et al. The results of suddendeafness treatment[J]. Otolaryngol Pol，1998，52(5):707-712.
[8]Schweinfurth JM，Cacace AT. Cochlear ischemia induced by circulating ironparticles under magnetic control: an animal model for sudden hearing loss[J].Am J Otol，2000，21(5):636-640.
[9]Sauvaget E，Kici S， Petelle B， et al. Vertebrobasilar occlusive disorderspresenting as sudden sensorineural hearing loss[J]. Laryngoscope，2004，114(2):327-332.
[10]Kazunori T，Temyuki H，Yasuhiro K， et al. Bilateral deafness as a prodromalsymptom of basilar artery occlusion[J]. J Neurol Sci，2002，193(2):147-150.
[11]李爱华，吴意.突发性耳聋患儿血液流变学指标检测及分析[J].实用预防医学,2010，17(11):2296-2297.
[12]季念英.突发性耳聋的病因研究进展[J].济宁医学院学报，2006，29(3):87-89.
[13]刘铤.梅尼埃病和突发性聋诊治的若干问题[J].中华耳鼻咽喉科杂志，1997，32（2）:124-125.
[14]郑金秀，陈国春，庄金梅，等.36例突发性耳聋伴眩晕临床分析[J].福建医学杂志，2001，23(5):111-112.
[15]俸家富，李冰.突发性耳聋患者外周血T细胞亚群的变化[J]，江西医学检验，2003，21(1):3-4.
[16]Solares CA，Tuohy VK.ELISPOT determination of interferon gamma T-cellfrequencies in patients with autoimmune sensorineural hearing loss[J]. Methodsin Molecular Biology，2005，302(3):253-260.
[17]Cadoni G，Fetoni AR，Agostino S，et al. Autoimmunity in sudden sensorineuralhearing loss：possible role of anti-endothelial cell auto antibodies[J]. ActaOtolaryngol Suppl，2002，122(5):30-33.
[18]Hardin MF， Barker M， Neis PR. Sensorineural hearing loss as the resultof cliff jumping[J]. J Ark Med Soc，2003，99:327-330.
[19]Foster PK，Luebke AE. A model for perilymphatic fistula induced hearing lossin the guinea pig cochlea[J]. Hearing Research，2002，167(1):175-179.
[20]Ottaviani F, Cadoni G, Marinelli L,et al. Anti0-endothelial auto-antibodiesin patients with sudden hearing loss[J]. Laryngoscope，1999，109(7):1084-1087.
[21]Patscheke JH， Arndt J， Dietz K， et al. The prothrombin G20210A mutationis a risk factor for sudden hearing loss in young patients[J]. Thromb Haemost，2001，86:1118-1119.
[22]Rudack C，Langer C，Junker R. Platelet GPIaC807T polymorphism is associatedwith negative outcome of sudden hearing loss[J]. Hearing Research，2004，191(1):41-48.
[23]柴峰.王永钦教授辨证论治暴聋经验[J].中华实用中西医杂志，2003,16（11）:1636.
[24]林祖辉，叶杰清，徐璐洁.辨证分型治疗突发性耳聋56例观察[J].实用中医药杂志，2003,19（3）：117-119.
[25]赵俐，海波.中西医结合治疗突发性耳聋疗效观察[J].内蒙古中医药，2003，22（6）：19.
[26]戈言平，沈强，徐锦.突发性耳聋纯音测听、经颅多普勒检查与中医证型相关性研究[J].中国中医急症，2009,18（5）：748—750.
[27]葛灏.突发性耳聋听力损失程度与中医辨证分型关系初探.南通医学院学报[J]，1999，19（2）：231.
[28]Kanemaru S， Fukushima H， Nakamura H， et al. Alpha-interferon for thetreatment of idiopathic sudden sensorineural hearing loss[J].Eur ArchOtorhinolaryngol，1997，254(3):158-162.
[29]王铁，苏崇周,王尔琴等.突发性耳聋三种药物疗效分析[J]，中国中西医结合耳鼻咽喉科杂志1995，3（2）:5-7.
[30]刘志慧，杨静波，李玲.对突发性耳聋病因及治疗的评估[J].航空航天医药，1998，9(3):154-155.
[31]李德炳，冯勇，王槐富.突发性耳聋治疗的新进展[J].听力学及言语疾病杂志，2004，12(5):349-351.
[32]Dodson KM，Sismanis A. Intratympanic perfusion for the treatment of tinnitus[J].Otolaryngol Clin North Am，2004，37(5):991-1000.
[33]Hoffmann F,Beck C,Schutz A,et al. Ginkgo extract EGb761(tenobin)/HAESversus naftidrofury/(Dusodril)/HAES. A randomized study of therapy of suddendeafness[J]. Laryngorhinootologie,2001,121:579-584.
[34]胡红.川芎嗪治疗突发性耳聋72例报告[J].咸宁学院学报(医学版)，2006，20（2）：151-153.
[35]赵健林.葛根素治疗突发性耳聋30例疗效观察[J]，山西医药杂志，2010，（39）8：748-749.
[36]郭丽,赵玉林,李小萍，等.复方丹参注射液治疗突发性耳聋66例[J].中医研究，2006，（19）5：40-41.
[37]栗珂，李秀敏，黄铁强.脉络宁治疗突发性耳聋临床观察[J].黑龙江中医药，2002，（3）：24.
[38]Zivic L，Zivic D，Stojanovic S. Sudden hearing loss our experience intreatment with vasoactive therapy [J]. Srp Arh Celok Lek，2008，136(3):91-94.
[39]邓基波，孙奉乾.前列腺素E脂微球制剂治疗突发性耳聋的疗效观察[J].中日友好医院学报，2006,20（1）：38-39.
[40]张洪颖.长春西汀注射液治疗脑梗死疗效观察[J].中国误诊学杂志，2008，8(12):2859-2860.
[41]孙洁，吕义美.突发性耳聋的药物治疗[J].中国基层医药，2004，11(6):761.
[42]沈峰.长春西汀治疗突发性耳聋的疗效观察[J].中国全科医学，2010,11:1238.
[43]王卫国，许荣，姚茂军.突发性耳聋的早期治疗及超氧化物歧化酶和丙二醛检测[J].中华耳鼻咽喉科杂志，2002，37(4):274-276.
[44]骆文龙，刘世喜，梁传余，等.三磷酸胞苷加地塞米松等综合治疗突发性耳聋的临床观察[J].中华耳鼻咽喉科杂志，2002，37（4）:277-279.
[45]张仲华，师秀萍，桂训海.应用单唾液酸四己糖神经节苷脂联合巴曲酶治疗突发性耳聋的临床研究[J].承德医学院学报，2009,26（1）：28-30.
[46]徐蓓,邵健.甲钴胺的临床应用[J].现代中西医结合杂志，2006,15(7):972-973.
[47]杨长君.甲钴胺治疗突发性耳聋的疗效观察[J].中国实用神经疾病杂志，2009,12（1）：97.
[48]Stokroos RJ，Albers FW，Schitm J，et al. The etiology of Idiopathic suddensensorineural hearing loss： Experimental herpes-simplex virus infection ofthe inner ear[J]. Am J Otol，1998，(19)4:447-452.
[49]李和清，任基浩，卢永德.突聋血清病毒特异性IgM抗体及NO检测的初步研究[J].中国耳鼻咽喉颅底外科杂志，2002，8（2）：133-134.
[50]Berrocal JR，Ramirez-Camacho R. Sudden sensorineural hearing loss: supporting theimmunologic theory. The Annals of Otology, Rhinology, and Laryngology，2002，111(11):989-997.
[51]Scherer EQ，Herzog M，Wangemann P. Endothelin-1-induced vasospasms of spiralmodiolar artery are mediated by rho-kinase-induced Ca(2+) sensitization ofcontractile apparatus and reversed by calcitonin gene-relatedPeptide[J].Stroke，2002，33(12):2965-2971.
[52]蒋春荣，殷团芳，卢永德，等.突聋与微循环障碍的关系[J].中国耳鼻咽喉颅底外科杂志，1999，5（1）：51-53.
[53]Thomas J Haberkamp， Hasan M Tanyeri.Management of idiopathic-suddensensorineural hearing loss [J].The American Journal of Otology，1999，20(5):587-595.
[54]Cole RR，Jahrsdoerfer RA. Sudden hearing loss:an update[J]. The AmericanJournal of Otology，1988，9(3):211-215.
[55]Minoda R， Masuyama K， Habu K， et al. Initial steroid hormone dose in thetreatment of idiopathic sudden deafness [J]. American Journal of Otology，2000，21(6):819-825.
[56]Herr BD， Marzo SJ. Intratympanic steroid perfusion for refractory suddensensorineural hearing loss[J]. Otolaryngol Head Neck Surg，2005，132(4):527-531.
[57]周杰，罗仁瀚,黄云声.龙胆泻肝汤加减治疗突发性耳聋30例临床观察[J].长春中医药大学学报,2009,25(1):120-121.
[58]李可.李可老中医急危重症疑难病经验专辑[M].太原:山西科学技术出版社，2002:274-275.
[59]杜长河.补阳还五汤加减治疗突发性耳聋55例[J].实用中医药杂志,2007，23（10）：640.
[60]沈映群.中药药理学[M].上海：上海科学技术出版社，2000:136.
[61]朱天民，孙宏，金荣疆.活血化瘀方药对突发性耳聋患者BAEP及TCD脑血流参数的影响[J]，中国中西医结合杂志，2006,26（8）:740-742.
[62]郭伟，吴志学，闻留瑞.耳聋复聪汤为主治疗突发性耳聋30例临床观察[J].江苏中医药，2002,23(11):39.
[63]李彦华，邹广华.补肾活血法治疗突发性耳聋36例疗效观察[J].新中医，2008,40（12）：58-59.
[64]张弢.行气复聪方治疗突发性耳聋遗留耳鸣的THI评价[J].上海中医药杂志，2010，44(8):41-43.
[65]孔敏.龙胆泻肝汤加活血药治疗突发性耳聋临床观察[J].辽宁中医药大学学报，2009,11（2）：108-109.
[66]罗巧云，周和平，吕新莲,等.氦—氖激光血管内照射治疗Ⅱ型糖尿病临床和血液流变学的观察[J].中华理疗杂志,1997，20(2):95-98.
[67]郭桂荣，张淑萍，周志霞，等.低能量氦—氖激光血管内照射治疗高脂血症40例[J].中国疗养医学，1998，7(1):15-17.
[68]荆忱，戴成祥，刘瑞云，等.低能量氦—氖激光血管内照射治疗冠心病60例[J].中华理疗杂志,1997，20(2):89-91.
[69]魏万林，林毅勇，徐秋玲，等.低能量氦—氖激光血管内照射治疗动脉栓塞性剧痛一例[J].中华理疗杂志,1997，20(1):63.
[70]高平青，姚富通，李虹，等.氦—氖激光血管内照射并脉络宁治疗急性脑梗塞临床研究[J].中华理疗杂志,1996，19(2):99-101.
[71]谭其仁，姜乃明，夏军，等.低强度氦—氖激光兔耳血管内照射对微循环障碍的影响[J].中华理疗杂志,1996，19(3):141-143.
[72]韦兴昌，刘开祥，俸军林，等.低能量氦—氖激光血管内照射疗法[J].中华理疗杂志,1995，18(1):45-48.
[73]齐杰，张素杰，朴永芬.低能量He-Ne激光血管内照射治疗心脑血管病20例[J].中国疗养医学,1996，5(1):42-44.
[74]孙曰升，周勇，王桂兰.氦氖激光血管内照射综合治疗快速性心律失常60例观察[J].中华理疗杂志,1998，21(6):365-367.
[75]丁桂才，王祝庆.低能量He-Ne激光血管内照射治疗外伤后头痛26例[J].中国疗养医学,1998，7(2):30-31.
[76]樊凌冰.高压氧治疗突发性耳聋的临床观察[J].青海医药杂志，1997，27(1):32-33.
[77]谢鼎华，杨伟炎.耳聋的临床与基础[M].长沙:湖南科学技术出版社，2004:246-251.
[78]陈学华，宁文静，吴世安.100例突发性耳聋疗效分析[J].实用医技杂志，2008，15(5):658-659.
[79]徐小军，赵志刚，李燕.突发性耳聋的治疗方法进展[J].医药导报，2000，19（6）:584-586.
[80]兰志红，邹朝福.突发性耳聋的疗效探索[J].中国中西医结合耳鼻咽喉科杂志，2002，10（2）:58-60.
[81]罗思瑾.药物、高压氧、体外反搏治疗突发性耳聋的疗效分析[J].暨南大学学报(医学版)，1999，20(2):99-102.
[82]马向明，任莉贇.针刺为主治疗突发性耳聋28例临床观察[J].中国中医药科技,2010,17（1）:31.
[83]范新华，丁亚南，常向辉.针灸与药物治疗突发性耳聋疗效对比观察[J].中国针灸，2010，30（8）:630-632.
[84]谢强，任元元，李唯钢.转移兴奋灶针灸法为主治疗感音神经性耳鸣的临床观察[J].实用中西医结合临床，2009，9(3):15-21.
[85]罗仁瀚,周杰,黄云声,等.电针治疗突发性耳聋疗效对照观察[J].中国针灸,2009,29（3）:185-187.
[1]Umemura K，Kohno Y，matsuno H，et al．A mew model for photochemically inducedthrombosis in the inner ear microcirculation and the use of hearing loss as ameasure for mirocircularory disorders[J].Eur Arch Otorhinolaryngol,1990，248：105-108．
[2]Jin K,Yasuhiro I,Kaoru O,et al.Effect of single-drug treatment onidiopathicsudden sensorineural hearing loss[J].Auris Nasus Larynx,2003,30(2):123-127.
[3]张琳，马峰，方亚伦，等，高血压病中医辨证与甲襞微循环变化的关系[J].微循环学杂志，1995，5（1）：39.
[4]黄焱明，邓水平，何世银.高血压病中医辨证与红细胞变形性关系的研究[J].四川中医，1993，11（2）：11.
[5]郑峰，胡世云，孙光.Ⅱ期高血压病中医证型与血浆内皮素一氧化氮关系的临床研究[J].福建中医药，2000，31（2）：5.
[6]王玉林，董明敏.耳蜗微循环研究进展[J].微循环学杂志，2002，12（2）：35-38.
[7] Hirano K,Ikeda K, Kawase T. Prognosis of sudden deafness with specialreference to risk factors of microvascular pathology[J]. Auris NasusLarynx,1999,26(2):111-115.
[8] Suzuki A,Ren T,Nuttulall al,et al.Age-related changes in cochlear bloodflow response to occlusion of anterior inferior cerebellar artery in mice[J].AnnOtol Rhinol Laryngol,1998,107(8):648—653.
[9]Goldwin B, Khan MJ, Seidman MD, et al. Sarthran preserves cochlear microcirculation and reduces temporary threshold shifts after noiseexposure[J].Otolaryngol Head Neck Surg,1998,118(5):576—583.
[10]Nagure M, Iwasaki S,Wu R,et al. Effects of corticosterod, contrast mediumand ATP on focal microcirculatory disorders of the cochlea[J]. Eur JPharmacol,1999,366(1):47-53.
[11] Penha R，O’Neill MA, Goyri O’Neill J,et al.Ultratructural aspects of themicrovasculture of the cochlea:the internal spiral network[J].Otolaryngol HeadNeck Surg,1999,120:725-729.
[12] Aird WC. Endothelium as an organ system[J]. Crit Care Med,2004,32(5):271.
[13]林琳,孙建和,黄德亮,等.胎儿耳蜗血管纹的扫描电镜观察[J].临床耳鼻咽喉科杂志,2005,19:157.
[14] Nakasima T, Naganawa S,Sone M,et al.Disorders of cochlear bloodflow[J].Brain Reviews,2003,43:17.
[15] Ohlemiler KK,Wright JS,Dugan LL.Early elevation of cochlear reactive oxygenspecies following noise exposure[J].Audiol Neurootol,1999,4:229.
[16] Losano G,Gattullo D,Pagliaro P.Myocardial neural and vascular aspects ofischemic preconditioning[J].Life Sciences,1996,59:185.
[17] Yang Gy,Betz AL. Reperfusion-induced injury to the blood-brain barrierafter middle cerebral artery occlusion in rats[J].Stroke,1994,25（1）：658.
[18] Brechtel P,Nuttal,Miller JM. Basal nitric oxide production in regulationof cochlear blood flow[J]. Hearing Research,1994,77:38.
[19]叶放蕾,董明敏.豚鼠基底动脉缺血一再灌注耳蜗损伤的实验研究[J].郑州大学学报(医学版),2002,37:320.
[20] Opremcak EM，Bruce RA，Lomeo MD，et al．Radial optic neurotomy for centralretinal vein occlusion：a retrospective pilot study of11consecutivecases[J].Retina，2001，21：408-415.
[21]吴昊，周维镕．豚鼠耳蜗微循环障碍模型的实验研究[J].南通医学院学报，2003，23：246-248．
[22]胡博华，周娜．不同程度的阻断内耳供血对耳蜗微循环和耳蜗生理功能的影响[J].军医进修学院学报，1999，20：179-181．
[23]罗志强，孔维佳，张先锋，等．三氯化铁诱导建立基底动脉栓塞动物模型[J]．临床耳鼻咽喉科杂志，2003，17（6）：364-366．
[24]Takeshita T，Iwasaki S，Nagura M，et al．Photochemically induced double lateralwall lesions in the guinea pig cochlea[J]．Acta Otolaryngol，2003，123：355-361．
[25] Scheibe F，Haupt H，Grunert H．Laser Doppler measurements of inner ear bloodflow during experimental thrombosis of cochlear blood vessels in the guineapig[J].Eur Arch Otorhinolaryngol，1997，254：86-90．
[26]王玉林，董明敏.光化学诱导豚鼠耳蜗微循环障碍的实验研究[J]，中国微循环，2002，6（6）：338-340.
[27]何景春，周维镕，严之曼等.光化学法诱导豚鼠耳蜗微循环障碍模型，中国耳鼻咽喉头颈外科[J]，2007，14（6）：361-364.
[28]Kavak li A,Sahna E, Parlakpinar H, et al.The effects of melaton in on focalcerebral ischem iareperfusion model[J].Saudi Med J,2004,25(11):1751-1752.
[29]席恺,董明敏．α-硫辛酸对豚鼠缺血-再灌注耳蜗损伤的防治作用，郑州大学学报(医学版)[J]，2005，40（2）：268-270.
[30]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：1237.
[31]范金茹，等.参附注射液与柴胡注射液抗脂质过氧化的作用[J].中成药，1991，13（2）：25.
[32]太田康幸.柴胡皂苷增强糖皮质激素作用的机制[J].国外医学·中医中药分册，1981，3（3）：58.
[33]朱江，蔡德海，芮菁.栀子的抗炎镇痛作用研究[J].中草药，2000，（31）3：198.
[34]王本祥主编.现代中药药理学[M].第1版.天津：天津科学技术出版社，1997，1107.
[35]陈长勋，金若敏.钩藤碱对血小板聚集和血栓形成的抑制作用[J].中国药理学报，1992，13（2）：126-130.
[36]胡锦官，顾健，王志旺.石菖蒲及其有效成分对中枢神经系统作用的实验研究[J].中药药理与临床，1999，15（3）：19.
[37]李明亚，陈红梅.石菖蒲对行为绝望动物抑郁模型的抗抑郁作用[J].中药材，2001，24（1）：40.
[38]张信岳，郑高利，寿燕，等.石菖蒲的益智和抗惊厥作用研究[J].浙江中医学院学报,1999,23(2):46.
[39]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：416.
[40]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：1237.
[41]刘军，吴卫平，匡培根.丹参在缺血再灌注时的神经保护作用及成纤维细胞生长因子上调[J].中国神经免疫学和神经病学杂志，1996，3（4）：225.
[42]李菊，杨期东.微透析技术和丹参注射液对Wistar鼠脑缺血时海马细胞外液兴奋性氨基酸释放的影响[J].中风与神经疾病杂志，1995，12（5）：258.
[43]柯朝阳，吴展元，肖伯奎.丹参、辅酶Q10对低氧供豚鼠动脉血气及耳蜗电图的影响[J].实用儿科临床杂志，2000，15（3）：132.
[44]蔡庆，刘红巾.碱性成纤维细胞生长因子和丹参对反复高Gz暴露大鼠脑组织中Bcl-2和P53mRNA表达的影响.空军总医学院学报[J]，2000，16（3）：130.
[45]吴卫平，姜树军，张小澍. C-Jun、bFGF、和HSP70在丹参改善颞叶缺血大鼠空间记忆障碍中的变化[J].卒中与神经疾病，1999，6（4）：193-195.
[46]黄青青，杨镛，万林骏，等.复方丹参注射液对兔内毒素休克的防治作用[J].中国危重病急救医学,1998，10（4）：227.
[47]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：170.
[48]Li M, Handa S, Ikeda Y, et al.Specific inhibiting characteristics oftertramethylpyrazine,one of active ingredients of the Chinese herbal medicine‘Chuanxiong,’on platelet thrombus formation under high shear rates[J]. ThrombRes,2001,104(1):15-28.
[49]范立，吕爱刚.川芎嗪对离体兔血管平滑肌和豚鼠盲肠带的作用[J].新药与临床，1997，16（4）：211.
[50]曲友直，高国栋，赵燕玲.川芎嗪、黄芪合用对脑缺血再灌注后脑组织C-Jun蛋白表达影响[J].安徽中医学院学报，2000，19（6）：256.
[51]Shih YH, Wu SL, Chiou WF, et al. Protective effects oftetramethylpyrazine on kainite-induced excitotoxicity in hippocampal culture.Neuro report[J],2002,13(4):515-519.
[52]蔡晨，等.川芎嗪对烫伤豚鼠血管内皮细胞保护作用的实验研究[J].中国烧伤创疡杂志，2000，（4）：11.
[53]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：1504—1507.
[54]Zhang GC.Antioxidation of pueraria lobata isoflavones[J].CA,1999,130:105.
[55]洪俐.葛根的研究进展概述.基层中医药杂志[J]，2000，14（2）：52.
[56]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：657-661.
[57]金鸣，李金荣.红花水溶性成分抗氧化作用的研究[J].心肺血管病杂志，1998，17（4）：277.
[58]张明霞，赵惠.红花对小鼠抗应激能力的影响[J].中国中医药信息杂志，1999，6（1）：26.
[59]马亚兵.枳壳的胃肠作用及炮制前后的变化[J].中药药理与临床，1996，12（6）：28.
[60]明彩荣，刘海星，井欢.中药抗过敏反应的显微微分干涉显微镜观察[J].中医药学刊，2001，19（3）：275.
[61]杨俊业，熊天琦.夜交藤煎剂的镇静催眠作用[J].华西医科大学学报，1990，21（2）：175.
[62]黄树莲，陈学芬.首乌藤降脂作用的实验研究[J].中草药，1991，22（3）：117.
[63]薛建欣，严永清，蒋莹.黄芪、归尾、香附、川芎、赤芍等配伍对“血瘀”大鼠血液流变学的影响.中国中药杂志，1994,19（2）:108.
[64]王本祥主编.现代中药药理与临床[M].天津:天津科技翻译出版公司，2004，6月第1版：428-429.
[65]包金凤，杨永健.甘草次酸钠对大鼠化学性腹膜炎的影响[J].中国药理学报，1997，18（3）：277.
[66]Kenzo Ohtsuki,Sanae Nakamura, Yoshihito Shimoyama, et al. A96-kDaglycyrrhizin-binding protein(gp96) from soybeans acts as a substrate for caseinkinase Ⅱ, and is highly related to lipoxygenase3[J]. The Journal ofBiochemistry,1995,118(6):1145-1150.
[67]Harada S, Karino A, Shimoyama Y,et al.Identification ofglycyrrhizin-binding protein kinase as casein Ⅱ and characterization of itsassociated phosphate acceptors in mouse liver[J].Biochem Biophys ResCommum,1996,227(1):102-109.
[68]句海松，忻文娟.甘草类黄酮对脂质过氧化和活性氧自由基的作用[J].药学学报，1998，24（11）：807.
[69]刘蕊、安杨.突发性耳聋证治的现代文献研究[J].新疆中医药，2009，27（3）：11-13.
[70]王高华，黄从新.青年抑郁障碍患者的血浆血小板因子4浓度[J].微循环学杂志，2003，13（2）：36-39.
[71]杨斌，叶兰仙，刘建斌，等.抑郁症患者血小板膜表面P-选择素的表达研究[J].中国神经精神疾病杂志，2005，31（1）：37-39.
[72]Delisi JS, Konopka JM, Russell K, et al. Platelet cytosolic calciumhyperresponsivity to serotonin in patients with hypertension and de-pressivesymptoms[J]. Biological Psychiatry,1999,45(8):1035-1041.
[73]钱欣梅,张向阳.氧自由基与耳聋[J].济宁医学院学报,2003，26(2):67.
[74]Coakun O, Ocakei A, Bayraktaroglu T, et al. Exercise training prevents andstreptozotocin-induced oxidative stress and β-cell damage in rat pancieas[J].Tohoku J Exp Med,2004,203(3):145-154.
[75]杨小莹，马世平，瞿融.柴胡提取物对慢性应激抑郁大鼠脂质过氧化及淋巴细胞增殖的影响[J].中国药科大学学报，2007，38（5）：442-445.
[76]Wichers M, Maes M. The psychoneuroimmuno-pathophysiology ofcytokine-induced depression in humans[J]. Int J Neuropsychopharmacol,2002,5(4):375-388.
[77]季宁东，李娟好，李明亚.石菖蒲提取液的抗抑郁作用及柴胡皂苷对其作用的影响[J].南京医科大学学报，2006，26（12）：1203-1206.
[78]王宝亮，苏谨.龙胆泻肝汤联合巴曲酶治疗突发性耳聋30例疗效观察[J].中国实用神经疾病杂志，2010，13（3）：79—80.
[79]Achen MG, Stacker SA.The vascular endothelial growth factor family proteinswhich guide the development of the vasculature[J]. Int J ExpPath,1998,79:255-265.
[80]李蕾，刘开祥. VEGF在缺血性脑血管病中研究进展[J].牡丹江医学院学报，2010，31（5）：69-71.
[81]Tuomisto TI, Rissanen TT, Vajanto I, et al.HIF-VEGF-VEGFR-2, TNF alpha andIGF pathways are up regulated in critical human skeletal muscle ischemia asstudied with DNA array[J].Atherosclerosis,2004,174:11-20.
[82]郭敬，王烈.血管内皮生长因子与治疗性血管生成研究进展[J].福州总医院学报，2010，17（1）：56-58.
[83]Shen F, FanY, SuH, et al. Adeno-associated viral vector-mediated hypoxia-regulated VEGF gene transfer promotes angiogenesis following focal cerebralischemia in mice[J]. Gene Ther,2008,15(1):30-39.
[84]Huang YF, Zhuang SQ, Chen DP, et al. Angiogenesis and its regulatory factorsin brain tissue of neonatal rat hypoxic-ischemic encephalopathy[J]. ZhonghuaErKe Za Zhi2004,42(3):210-214.
[85]梅爱农,姜亚平.缺血性脑血管病与血管生成[J].国外医学脑血管疾病分册,2004,12(1):67-70.
[86]Sengupta S, Toh SA, Sellers LA, et al. Modulating angiogenesis: the yin andthe yang in ginseng[J].Circulation,2004,110(10):1219.
[87]]王建湘,王行宽,尹树忠.心痛治肝法“心痛灵Ⅱ号”对冠心病心绞痛患者血脂代谢及内皮功能的影响[J].中国中医急症,2003,12(5):392.
[88]王文,王宗仁,张金洲,等.芪丹通脉片对大鼠缺血心肌的血管新生作用与机制研究[J].中国中医急症,2007,16(2):190.
[89]王振涛,韩丽华,朱明军,等.黄酮皂苷类中药对促心肌梗死后大鼠缺血心肌血管新生作用及相关生长因子表达的影响[J].中西医结合心脑血管病杂志,2006,4(4):305.
[90]]张喜成,何延政,李跃武,等.补肾生血胶囊动员骨髓内皮祖细胞促进缺血肢体血管新生的研究[J].江苏中医药,2006,27(3):57
[91]Yang ZJ, Bao WL, Qiu MH, et al. Role of vascular endothelial growth factorin neuronal DNA damage and repair in rat brain follow-ing a transient cerebralischemia［J. J Neurosci Res,2002,70(2):140-149.
[92]Bottcher R T，Niehrs C．Fibroblast growth factor signaling during earlyvertebratedevelopment［J］．Endocrine Rev，2005，26（1）：63－77．
[93］Damico F M．Angiogenesis and retinal diseases［J］．Arq Bras Oftlmol，2007，70（3）：547－553．
[94]王晶，尹丽娅，李庚山，等．bFGF对缺氧条件下培养的人脐静脉内皮细胞增殖影响［J］．武汉大学学报：医学版，2002，23（4）：298－300．
[95]Hall H．Modified fibrin hydrogel matrices：both，3D-scaffolds and local andcontrolled release systems to stimulate angiogenesis［J］．Curr Pharm Des，2007，13（35）：3597－3607．
[96]Koivisto H，Hyvarinen M，Stromberg AM，et al．Cultures of human embryonicstem cells：serum replacement medium or serum-containing media and the effectof basic fibroblast growth factor［J．Reprod Biomed Online，2004，9（3）：330－337．
[97]Cutroneo K R．Gene therapy for tissue regeneration［J］．Cell Biochem，2003，88（2）：418－425．
[98]Coumoul X，Deng C X．Roles of FGF receptors in mammalian development andcongenital diseases［J］．Birth Defects Res C Embryo Today，2003，69（4）：286－304．
[99]Mueller KL, Jacques BE, Kelley MW. Fibroblast growth factor signalingregulates pillar cell development in the organ of Corti[J]. J Neurosc,2002,22(21):9368-9377.
[100]Carmeliet P. Mechanisms of angiogenesis and arteriogenesis［J］. NatureMedicine,2000,6(4):389-395.
[101]郑鸣，王丰，梁书锋.豚鼠耳蜗缺血再灌注损伤后iNOS活性与细胞凋亡关系[J].中国临床解剖学杂志，2006.24（2）：190-193.
[102] Ruan RS, Leong SK, YeohKH. Effects of nitric oxide on normal and ischemiccochlea of the guinea pig[J]. Exp Neuro,l2001,169(1):200-207.
[103]胡安，金晓杰.耳蜗微循环缺血与再灌注损伤的研究进展[J].听力学及言语疾病杂志，2008，1（5）：437-438.
[104]熊巍，邱建华.耳蜗的缺血/再灌注损伤研究进展[J].国外医学耳鼻咽喉科学分册，2005,29（4）：201-203.
[105]Cash D,Beech JS,Rayne RC,et al. Neuroprotective effect of aminoguanidineon transient focal ischaemia in the rat brain[J].Brain Res,2001,29(1):91-103.
[106]叶放蕾,董明敏.豚鼠基底动脉缺血一再灌注耳蜗损伤的实研究[J].郑州大学学报(医学版),2002,37:320.
[107]Lepore DA.Nitric oxide synthase-independent generation of nitric oxide inmuscle ischemia-reperfusiion injury[J].Nitric Oxide，2000,4(6):541-545.
[108]Seidman MD,Bai U,Khan MJ,et al.Mitochondrial DAN deletion associated withaging and presbyacusis[J].Arch Otolaryngol Head Neck Surg，1997,123(10):1039-1045.
[109]Berges A,Van Nassauw L,Bosmans J,et al.Role of nitric oxide and oxidativestress in ischemic myocardial injury and preconditioning[J]. ActaCardiol,2003,58(2):119-132.
[110]邓芬,胡长林,谢运兰.丹红注射液对大鼠局灶脑缺血后一氧化氮合酶的影响[J].中西医结合心脑血管病杂志，2006，4（10）：880-881.
[111]Moncada S,Palmer RM. Nitric oxide: physiology,pathophysiology andpharmacology[J]. Pharmacological Reviews,1991,43(2):109-142.
[112]Danielisova V, Nemethova M, Burda J.The protective effect ofaminoguanidineon cerebral ischemic damage in the rat brain[J]. Physiol Res,2004,53(5):533-540.
[113]陈山泉.血府逐瘀汤治疗冠心病心绞痛的临床研究[J].中医函授通讯,1999,18(6):48.
[114]洁西卡，桃李，章李军,等.诱导型一氧化氮合酶抑制剂研发现状[J].现代生物医学进展，2007，17（11）：1741-1743.
[115]Chambers AF，Matrisian LM．Changing views of the role of matrixmetalloproteinases metalloproteinases in metastasis［J］.J Natl Cancer Inst，1997，89(17):1260-1270．
[116]姜双，陈晓隆. ROP小鼠视网膜组织中MMP-2、VEGF的表达变化及意义[J].山东医药，2010，50（42）：28-29.
[117]Turpeenniemi-Hujanen T.Gelatinases(MMP-2and-9)and their naturalinhibitors as prognostic indicators in solid cancers[J].Biochimie,2005,87(3):287-297.
[118]Nagel S, Su Y, Horstmann S,et al. Minocycline and hypothermia forreperfusion injury after focal cerebral ischemia in the rat: effects on BBBbreakdown and MMP expression in the acute and subacute phase [J]. Clin Chem,2010,56:21-33.
[119] Pfefferkorn T,Rosenberg GA.Closure of the blood-brain barrier by matrixmetalloproteinase inhibition reduces rtPA-mediated mortality in ce-rebralischemia with delayed reperfusion[J].Stroke,2003,34(8):2025-2030.
[120]张会超，韩丽华，王振涛，等.益气活血方对心梗后左室重构大鼠心肌基质金属蛋白酶-2及转化生长因子β1的影响[J].中成药，2010,32（11）：1871-1874.
[121]伍尚敏，张蔓菁，刘宾,等.地塞米松对血管瘤内皮细胞分泌MMP-2的影响[J].中国美容医学，2010，19（11）：1645-1649.
[122]陈彩萍,过晶.心通口服液对冠心病心绞痛患者血MMP-2、MMP-9的影响[J].实用临床医药杂志，2010,14（21）：87-89.
[123]雷燕，杨静，赵浩，等.人参三七川芎提取物延缓衰老小鼠血管老化的实验研究[J].中国中西医结合杂志，2010,30（9）：946-951.
[124]] Harkness KA, Adamson P, Sussman JD, et al. Dexamethasone regulation ofMatrix metalloproteinase expression in CNS vascular endothelium [J]. Brain,2000,123(4):698-709.
[125]杨爱东,李文雯,汪东颖，等.清热燥湿方对急性肺损伤大鼠肺组织基质金属蛋白酶2及其抑制因子基因表达的影响[J].中国中医药信息杂志,2010,17(8):27-29.
[126] Jialong Guo,Kailun Zhang,Yanmei Ji, et al. Effects of ethyl pyruvate onmyocardial apoptosis and expression of Bcl-2and Bax proteins after ischemiareperfusion in rats［J］.Huazhong Univ Sci Technol Med Sci,2008,28(3):281-3.
[127]Li JS, Zhang W,Kang ZM, et al. Hyperbaric oxygen preconditioningreduces ischemia-reperfusion injury by inhibition of apoptosis viamitochondrial pathway in rat brain［J］. Neuroscience.2009,159(4):1309-1315
[128] Cardoso CM, Almeida LM, Custodio JB. Protection of tamoxifen againstoxidation of mitochondrial thiols and NAD(P)H underlying the permeabilitytransition induced by prooxidants[J]. Chem Biol Interact,2004,148:149-161.
[129]Lawrence MS, Mclaughlin JR, Sun GH, et al. Herpes simplex viral vectorsexpressing Bcl-2are neuroprotective when delivered after a stroke[J].J CerebBlood FlowMetab,1997,17:740-744.
[130] Antonawich FJ,Federoff HJ,Davis IN.Bcl-2trannsduction, using a herpessimplex virus amplicon, protects hippocampal neurons from transient globalischemia[J].Exp Neurol,1999,156:130-137.
[131] Jayasurya A,Dheen ST,Yap WM,et al. Inducible nitric oxide syn-thase andBc1-2expression in nasopharyngeal cancer: correlationwith outcome ofpatientsafter radiotherapy[J]. Int JRadiatOncol BiolPhys,2003,56(3):837-845.
[132] Shimizu S，Nagayama T，Jin KL，et al.Bcl-2antisense treatment preventinduction of tolerance to focal ischemia in the rat brain［J］.Cereb Blood FlowMetab，2001，21(3):233-243.
[133]李浩,刘开祥.丹参酮ⅡA对鼠脑缺血再灌注损伤Bcl-2和caspase-3表达的影响[J].中国康复医学杂志,2008,23(8):691-693
[134]戴雍月,徐正衸,王万铁等.红花注射液对肺缺血再灌注损伤时细胞凋亡及Bcl-2和Bax基因的影响[J].中国动脉硬化杂志，2007,15（6）：415-419.
[135]许益笑，王万铁，徐正衸.葛根素对兔肺缺血再灌注损伤时Bcl-2、Bax及caspase-3的影响[J].温州医学院学报,2007,37（2）：126-129.
[136]胡永红，周爱民，田野，等.川芎嗪促进脑缺血耐受形成的作用及机制研究[J].中国中医基础医学杂志，2010,16（8）：671-674.
[137]樊蔚虹,姚建平,赵文景.柴胡疏肝散对卒中后抑郁大鼠海马组织Bcl-2,Bax蛋白表达的影响[J].中国实验方剂学杂志，2011.11（3）：181-183.
[138]Peng CH, TangTH,Huang CN, et al. Optosis induced by penta-acetyl geniposidein C6glioma cells is associated with JNK activation and Fas ligand induction[J].Toxicol Appl Pharmacol,2005,202(2):172-179.
[139]方永奇,匡忠生,谢宇辉,等.石菖蒲对缺血再灌注脑损伤大鼠神经细胞凋亡的影响[J].现代中西医结合杂志，2002，11(17):1647-1649.
[140]蒋建刚，吴基良，陈金和.甘草酸二铵对大鼠心肌缺血再灌注损伤后细胞凋亡的影响[J].微循环学杂志，2004,14（4）：31-33.