益气活血、化瘀通络之中药对肾间质纤维化的作用研究
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摘要
目的:肾间质纤维化(tubulointerstitial,TIF)是各种原因所致肾脏疾病进展至终末期肾功能衰竭的共同病理特征[1]。大量临床病理资料表明,各种原因引起的慢性肾功能损伤与肾小管间质纤维化病变的严重程度较与肾小球病变的关系更为密切。而细胞外基质(extracellular martix,ECM)的合成和降解失衡,造成其大量积聚是引起肾间质纤维化的主要原因[2]。临床上应用血管紧张素转化酶抑制剂(angiotensin-converting enzyme inhibitor,ACEI)、血管紧张素II受体拮抗剂(angiotensin II receptor antagonist,ARB)等药物,通过调节ECM合成、降解来防治TIF,但循证医学已证明,上述药物在血肌酐高于一定值时,对肾脏不但无保护作用,还能加重其损害,且药价偏高,因而使其应用受限。因此寻找具有多途径多靶点治疗作用、安全低成本的药物对肾间质纤维化的防治有重要的意义。本实验应用导师经验方龙甲参芪汤对单侧输尿管结扎(unilateral ureteral obsurtction,UUO)诱导的肾间质纤维化大鼠模型进行干预性治疗,并以ARB类药物缬沙坦(Valsartna)为对照,旨在通过观察龙甲参芪汤对实验动物梗阻侧肾脏组织形态学及转化生长因子β_1(transforming growth factorβ_1,TGF-β_1)、III型胶原(collagenIII,ColIII)及其受体的影响,探讨中药复方—龙甲参芪汤对肾间质纤维化的拮抗作用及可能的作用机制。
方法:本实验研究选用雄性SD大鼠40只,适应性饲养一周后,分别置代谢笼里取尿,测定尿蛋白及尿红细胞全部阴性,随机分为4组,假手术组(SHAM group)、模型组(UUO group)、缬沙坦组(UUOV group)、龙甲参芪汤(UUOL group)组,每组10只,除假手术组外,各组动物均结扎左侧输尿管并在术前一天灌胃给药,假手术组与模型组分别灌入等量的生理盐水,于结扎后第15天用全自动生化分析仪测各组大鼠的血肌酐(SCr)、尿素氮(BUN),杀检测肾/体比(Index),并取左侧肾脏行病理切片,光镜下观察各组大鼠肾间质及肾小管的变化情况,免疫组化法检测TGF-β_1、ColIII在肾间质及肾小管的表达情况,并用病理图像分析软件进行半定量分析。
结果:
1 Index、SCr、BUN检测结果。
实验结束时,左侧肾脏体积增大,大鼠Index模型组比假手术组明显升高(p<0.01),缬沙坦组及龙甲参芪汤组与模型组比较显著降低(p<0.05,p<0.05),其中缬沙坦组较龙甲参芪汤组略低,但两组间无统计学意义(p>0.05)。SCr测定结果显示,模型组明显高于假手术组(p<0.05),缬沙坦组、龙甲参芪汤组大鼠的SCr与模型组比较显著降低(p<0.05,p<0.05),其中缬沙坦组的SCr低于龙甲参芪汤组,但无统计学意义(p>0.05)。BUN测定结果显示,模型组大鼠BUN明显高于假手术组(p<0.05),缬沙坦组、龙甲参芪汤组与模型组相比显著降低(p<0.05,p<0.05),其中缬沙坦组BUN低于龙甲参芪汤组,但无统计学意义(p>0.05)。
2肾组织HE及天狼猩红染色显示:
模型组间质大量炎性细胞浸润和纤维组织增生,肾小管部分萎缩、管腔闭塞或扩张,肾小球数目明显减少,面积缩小,肾小囊黏连或扩张,小囊周围大量纤维组织增生。各治疗组间质可见多少不等的炎性细胞浸润,少量纤维组织增生,肾小管轻度扩张,部分肾小球轻度萎缩,小囊扩张,肾小球数量无明显改变。
3 TGF-β_1在肾组织的表达、分布及半定量分析结果:
TGF-β_1主要在梗阻肾近曲小管上皮细胞胞浆及间质细胞内呈强阳性表达,假手术组实验动物在肾小管上皮细胞内TGF-β_1蛋白为弱阳性,其余各组在肾小管上皮细胞、间质细胞及肾小球内表达显著增强,其中模型组较假手术组TGF-β_1的阳性面积及积分光密度值均呈显著性升高(p<0.01),缬沙坦组、龙甲参芪汤组与模型组相比,TGF-β_1的阳性面积及积分光密度均呈显著性下降(p<0.01,p<0.01),提示龙甲参芪汤、缬沙坦均可抑制梗阻侧肾组织TGF-β_1的表达,缬沙坦组优于龙甲参芪汤组,但无显著性差异(p>0.05)。
4 ColIII在肾组织的表达、分布及半定量分析结果:
ColIII主要表达于梗阻肾的间质细胞内。假手术组大鼠肾间质ColIII呈弱阳性表达,其余各组表达显著增强,为条索状或成片块状。其中,模型组与假手术组相比ColIII的阳性面积及积分光密度值呈显著性升高(p<0.01),缬沙坦组、龙甲参芪汤组与模型组相比,ColIII的阳性面积及积分光密度值均呈显著性下降(p<0.01,p<0.01),提示龙甲参芪汤、缬沙坦均可抑制梗阻侧肾组织中ColIII的组织表达,其中,缬沙坦组优于龙甲参芪汤组,但无显著性差异(p>0.05)。
5肾间质纤维化面积半定量分析结果
模型组肾间质纤维化程度较假手术组明显加重(p<0.01);缬沙坦组、龙甲参芪汤组与模型组比较,肾间质纤维化面积明显减少(p<0.01,p<0.05),提示龙甲参芪汤及缬沙坦均有拮抗肾间质纤维化的作用。其中,缬沙坦组优于龙甲参芪汤组,但无显著性差异(p>0.05)。
结论:
1龙甲参芪汤能降低血清肌酐、尿素氮,降低肾脏指数,保护肾功能。
2龙甲参芪汤能抑制TGF-β_1、ColIII在肾组织的表达,从而减少病变肾组织ECM成分的合成及积聚,延缓肾间质纤维化的进程。
3龙甲参芪汤对UUO大鼠肾间质纤维化的拮抗作用与缬沙坦基本相似。
Objective:Tubulointerstitial fibrosis (TIF) is a common pathological feature of most end-stage kidney disease. Numbers of clinical pathology observations had shown that tubulointerstitial injury played an more important role than glomerular damage on renal failure. It is the TIF’s cause that Disequilibrium of extracellular martix (ECM) composition and degradation lead to a great quantity ECM’s accumulation. Clinically,Angiotensin Converting Enzyme Inhibitor(ACEI)and Angiotensin II ReceptorAntagonist(ARB) etal are applied to prevent and cure TIF by adjusting synthesis and degrada -tion of ECM and transformation of myofibroblast(Myo-FB). However the medical science had testified that above-mentioned higher price medicines not only can’t protect kidney,but also might lesion it in Scr higher than certain numerus,Therefore, to search the medicine that have many channels, many targets, safe and low cost treatment, there is a world of significance to prevent and cure TIF. In the experiment, The TIF that unilateral ureteral obsurtction(UUO) rat model induce to are treated by advisor’s empirical formula- Longjiashenqi soup,and contrast with ARB- Valsartna to observe the change that Longjiashenqi soup effect observeexperimental animal Obstruct kidney’s transforming growth factorβ_1(TGF-β_1), collagenIII(ColIII) and histomorphology so as to approach Chinese medicinal formulae Longjiashenqi soup antagonism and mechanism of action in TIF.
Methods: Fourty female Sprague-Dawley rats were randomly divided into four groups(n=10), Shma-operation (SHAM), model(UUO), Longjiashenqi soup (UUOL) and Valsartna (UUOV). Each animal was ligated left ureter except SHAM group. Durgs were administered from the day before operation but the saline were used in SHAM and UUO groups by oral. Animals were sacrificed after twenty-one days the left kidneys were harvested for pathological study. Imm -unohistochemisyrt staining were used for expression of TGF-β_1, ColIII,and to apply patho-image analysis software to do semi-quantitative analysis.
Results:
1 The detection result on Index、SCr、BUN
1.1 At the end of experiment, left kidney volume is increasing,the Index on UUO group was higher than that on SHAM group(p<0.01);compared with the UUO group,The Index of UUOV,UUOL groups decreased marked ly (p<0.05, p <0.05). but there was not statistically significant between them(p>0.05).
1.2 The SCr of UUO group was higher than that on SHAM group(p<0.05);compared with the UUO group,the SCr of UUOV,UUOL groups decreased markedly(p<0.05, p<0.05). but there was not statistically significant between them(p>0.05).
1.3 The BUN of UUO group was higher than that on SHAM group(p<0.05);compared with the UUO group,the BUN of UUOV,UUOL groups decreased markedly(p<0.05, p<0.05). but there was not statistically significant between them(p>0.05).
2 The renal tissues were stained with HE and Tianlangxing -hong:There was no change in SHAM group.But in UUO group,interstitial macrophage and monocyte infiltra -tion,renal tubular atrophy,emphraxis or expand were shown. The number of glomeurli reduced markedly,and the area of glomeurli decreased.The renal capsule adhered or expanded. Fibrous tissues proliferated around the renal cpasule.While the lesions in treatment groups were slighter than that in UUO group.
3 The expression,distribution and semi-quantitative analysis of TGF-β_1: It was shown a weak expression for TGF-β_1 in SHAM group,while in other groups the expression of TGF-β_1 inereased. They can be seen in kidney proximal tubule, cellula epithelialis endochylema and interstitial cell. The Positive area and density of TGF-β_1 in UUO group was higher than that in SHAM group(p<0.01);compared with the UUO group,the Positive area and density of TGF-β_1in UUOV ,UUOLgroups decreased markedly(p<0.01,p<0.01). It means that Longjiashenqi soup and Valsartan can inhibit the expression of TGF-β_1 in obstructive kindye.The inhibition of UUOV group was stronger than that of UUOL group,but there was not statistically significant between them(p>0.05).
4 The expression, distribution and semi-quantitative analysis of ColIII: They can be seen in renal interstitium. It was shown a weak expression for ColIIIin SHAM group,while in other groups the expression of ColIII inereased.The Positive area and density of ColIII in UUO group was higher than that in SHAM grouP(p<0.01);compared with the UUO group,the Positive area and density of ColIII in UUOV, UUOLgroups decreased markedly(p<0.01,p<0.01). It means that Longjiashenqi soup and Valsartan can inhibit the expression of ColIII in obstructive kindye.The inhibition of UUOV group was stronger than that of UUOL group,but there was not statistically significant between them(p>0.05).
5 In UUO group,the area of fibrosis in UUO group were more serious than in Sham group; the area of fibrosis in UUOV, UUOL groups were decreased markedly compared with the UUO group (p<0.01,p<0.05).It means that Longjiashenqi soup and Valsartan can inhibit fibrosis in obstructive nephro pathy. Among them , the inhibition of UUOL group was stronger than that of UUOVgroup,but there was not statistically significant between them(p>0.05).
Conclusion:
1 Longjiashenqi soup can degrade serum creatinine and urea nitrogen, protect renal function.
2 Longjiashenqi soup can restrain expression of TGF-β_1, ColIII in nephridial tissue so as to reduce pathological changes synthesis and accumulation of ECM, finally, it can delay TIF’s proceeding.
3 Longjiashenqi soup’s intervention effect to TIF of the UUO rat is similar to Valsartan’s.
方法:本实验研究选用雄性SD大鼠40只,适应性饲养一周后,分别置代谢笼里取尿,测定尿蛋白及尿红细胞全部阴性,随机分为4组,假手术组(SHAM group)、模型组(UUO group)、缬沙坦组(UUOV group)、龙甲参芪汤(UUOL group)组,每组10只,除假手术组外,各组动物均结扎左侧输尿管并在术前一天灌胃给药,假手术组与模型组分别灌入等量的生理盐水,于结扎后第15天用全自动生化分析仪测各组大鼠的血肌酐(SCr)、尿素氮(BUN),杀检测肾/体比(Index),并取左侧肾脏行病理切片,光镜下观察各组大鼠肾间质及肾小管的变化情况,免疫组化法检测TGF-β_1、ColIII在肾间质及肾小管的表达情况,并用病理图像分析软件进行半定量分析。
结果:
1 Index、SCr、BUN检测结果。
实验结束时,左侧肾脏体积增大,大鼠Index模型组比假手术组明显升高(p<0.01),缬沙坦组及龙甲参芪汤组与模型组比较显著降低(p<0.05,p<0.05),其中缬沙坦组较龙甲参芪汤组略低,但两组间无统计学意义(p>0.05)。SCr测定结果显示,模型组明显高于假手术组(p<0.05),缬沙坦组、龙甲参芪汤组大鼠的SCr与模型组比较显著降低(p<0.05,p<0.05),其中缬沙坦组的SCr低于龙甲参芪汤组,但无统计学意义(p>0.05)。BUN测定结果显示,模型组大鼠BUN明显高于假手术组(p<0.05),缬沙坦组、龙甲参芪汤组与模型组相比显著降低(p<0.05,p<0.05),其中缬沙坦组BUN低于龙甲参芪汤组,但无统计学意义(p>0.05)。
2肾组织HE及天狼猩红染色显示:
模型组间质大量炎性细胞浸润和纤维组织增生,肾小管部分萎缩、管腔闭塞或扩张,肾小球数目明显减少,面积缩小,肾小囊黏连或扩张,小囊周围大量纤维组织增生。各治疗组间质可见多少不等的炎性细胞浸润,少量纤维组织增生,肾小管轻度扩张,部分肾小球轻度萎缩,小囊扩张,肾小球数量无明显改变。
3 TGF-β_1在肾组织的表达、分布及半定量分析结果:
TGF-β_1主要在梗阻肾近曲小管上皮细胞胞浆及间质细胞内呈强阳性表达,假手术组实验动物在肾小管上皮细胞内TGF-β_1蛋白为弱阳性,其余各组在肾小管上皮细胞、间质细胞及肾小球内表达显著增强,其中模型组较假手术组TGF-β_1的阳性面积及积分光密度值均呈显著性升高(p<0.01),缬沙坦组、龙甲参芪汤组与模型组相比,TGF-β_1的阳性面积及积分光密度均呈显著性下降(p<0.01,p<0.01),提示龙甲参芪汤、缬沙坦均可抑制梗阻侧肾组织TGF-β_1的表达,缬沙坦组优于龙甲参芪汤组,但无显著性差异(p>0.05)。
4 ColIII在肾组织的表达、分布及半定量分析结果:
ColIII主要表达于梗阻肾的间质细胞内。假手术组大鼠肾间质ColIII呈弱阳性表达,其余各组表达显著增强,为条索状或成片块状。其中,模型组与假手术组相比ColIII的阳性面积及积分光密度值呈显著性升高(p<0.01),缬沙坦组、龙甲参芪汤组与模型组相比,ColIII的阳性面积及积分光密度值均呈显著性下降(p<0.01,p<0.01),提示龙甲参芪汤、缬沙坦均可抑制梗阻侧肾组织中ColIII的组织表达,其中,缬沙坦组优于龙甲参芪汤组,但无显著性差异(p>0.05)。
5肾间质纤维化面积半定量分析结果
模型组肾间质纤维化程度较假手术组明显加重(p<0.01);缬沙坦组、龙甲参芪汤组与模型组比较,肾间质纤维化面积明显减少(p<0.01,p<0.05),提示龙甲参芪汤及缬沙坦均有拮抗肾间质纤维化的作用。其中,缬沙坦组优于龙甲参芪汤组,但无显著性差异(p>0.05)。
结论:
1龙甲参芪汤能降低血清肌酐、尿素氮,降低肾脏指数,保护肾功能。
2龙甲参芪汤能抑制TGF-β_1、ColIII在肾组织的表达,从而减少病变肾组织ECM成分的合成及积聚,延缓肾间质纤维化的进程。
3龙甲参芪汤对UUO大鼠肾间质纤维化的拮抗作用与缬沙坦基本相似。
Objective:Tubulointerstitial fibrosis (TIF) is a common pathological feature of most end-stage kidney disease. Numbers of clinical pathology observations had shown that tubulointerstitial injury played an more important role than glomerular damage on renal failure. It is the TIF’s cause that Disequilibrium of extracellular martix (ECM) composition and degradation lead to a great quantity ECM’s accumulation. Clinically,Angiotensin Converting Enzyme Inhibitor(ACEI)and Angiotensin II ReceptorAntagonist(ARB) etal are applied to prevent and cure TIF by adjusting synthesis and degrada -tion of ECM and transformation of myofibroblast(Myo-FB). However the medical science had testified that above-mentioned higher price medicines not only can’t protect kidney,but also might lesion it in Scr higher than certain numerus,Therefore, to search the medicine that have many channels, many targets, safe and low cost treatment, there is a world of significance to prevent and cure TIF. In the experiment, The TIF that unilateral ureteral obsurtction(UUO) rat model induce to are treated by advisor’s empirical formula- Longjiashenqi soup,and contrast with ARB- Valsartna to observe the change that Longjiashenqi soup effect observeexperimental animal Obstruct kidney’s transforming growth factorβ_1(TGF-β_1), collagenIII(ColIII) and histomorphology so as to approach Chinese medicinal formulae Longjiashenqi soup antagonism and mechanism of action in TIF.
Methods: Fourty female Sprague-Dawley rats were randomly divided into four groups(n=10), Shma-operation (SHAM), model(UUO), Longjiashenqi soup (UUOL) and Valsartna (UUOV). Each animal was ligated left ureter except SHAM group. Durgs were administered from the day before operation but the saline were used in SHAM and UUO groups by oral. Animals were sacrificed after twenty-one days the left kidneys were harvested for pathological study. Imm -unohistochemisyrt staining were used for expression of TGF-β_1, ColIII,and to apply patho-image analysis software to do semi-quantitative analysis.
Results:
1 The detection result on Index、SCr、BUN
1.1 At the end of experiment, left kidney volume is increasing,the Index on UUO group was higher than that on SHAM group(p<0.01);compared with the UUO group,The Index of UUOV,UUOL groups decreased marked ly (p<0.05, p <0.05). but there was not statistically significant between them(p>0.05).
1.2 The SCr of UUO group was higher than that on SHAM group(p<0.05);compared with the UUO group,the SCr of UUOV,UUOL groups decreased markedly(p<0.05, p<0.05). but there was not statistically significant between them(p>0.05).
1.3 The BUN of UUO group was higher than that on SHAM group(p<0.05);compared with the UUO group,the BUN of UUOV,UUOL groups decreased markedly(p<0.05, p<0.05). but there was not statistically significant between them(p>0.05).
2 The renal tissues were stained with HE and Tianlangxing -hong:There was no change in SHAM group.But in UUO group,interstitial macrophage and monocyte infiltra -tion,renal tubular atrophy,emphraxis or expand were shown. The number of glomeurli reduced markedly,and the area of glomeurli decreased.The renal capsule adhered or expanded. Fibrous tissues proliferated around the renal cpasule.While the lesions in treatment groups were slighter than that in UUO group.
3 The expression,distribution and semi-quantitative analysis of TGF-β_1: It was shown a weak expression for TGF-β_1 in SHAM group,while in other groups the expression of TGF-β_1 inereased. They can be seen in kidney proximal tubule, cellula epithelialis endochylema and interstitial cell. The Positive area and density of TGF-β_1 in UUO group was higher than that in SHAM group(p<0.01);compared with the UUO group,the Positive area and density of TGF-β_1in UUOV ,UUOLgroups decreased markedly(p<0.01,p<0.01). It means that Longjiashenqi soup and Valsartan can inhibit the expression of TGF-β_1 in obstructive kindye.The inhibition of UUOV group was stronger than that of UUOL group,but there was not statistically significant between them(p>0.05).
4 The expression, distribution and semi-quantitative analysis of ColIII: They can be seen in renal interstitium. It was shown a weak expression for ColIIIin SHAM group,while in other groups the expression of ColIII inereased.The Positive area and density of ColIII in UUO group was higher than that in SHAM grouP(p<0.01);compared with the UUO group,the Positive area and density of ColIII in UUOV, UUOLgroups decreased markedly(p<0.01,p<0.01). It means that Longjiashenqi soup and Valsartan can inhibit the expression of ColIII in obstructive kindye.The inhibition of UUOV group was stronger than that of UUOL group,but there was not statistically significant between them(p>0.05).
5 In UUO group,the area of fibrosis in UUO group were more serious than in Sham group; the area of fibrosis in UUOV, UUOL groups were decreased markedly compared with the UUO group (p<0.01,p<0.05).It means that Longjiashenqi soup and Valsartan can inhibit fibrosis in obstructive nephro pathy. Among them , the inhibition of UUOL group was stronger than that of UUOVgroup,but there was not statistically significant between them(p>0.05).
Conclusion:
1 Longjiashenqi soup can degrade serum creatinine and urea nitrogen, protect renal function.
2 Longjiashenqi soup can restrain expression of TGF-β_1, ColIII in nephridial tissue so as to reduce pathological changes synthesis and accumulation of ECM, finally, it can delay TIF’s proceeding.
3 Longjiashenqi soup’s intervention effect to TIF of the UUO rat is similar to Valsartan’s.
引文
1 Bohle A,Muller GA,Wehrmanun M,et al. Pathogenesis of Chornic renal failure in the Primary glomerulopathies, renal Vasculopathies,and chronic interstitial nephritides.Kidney Int,1996,49(Suppl54):S2~S9
2 Kitamura K,Kangawa M,Kawamoto M,et al. Adrenome dullin:a novel hypotensive peptide isolated from human pheochromacytoma.Biochem BioPhys Res Commun, 1993,192:553~560
3 候凡凡.肾小管间质性疾病发病机理进展.国外医学内科分册,1991,18(8):361
4 Kuncio GS,Nilson EG,Haverty T.mechanisms of tubulionterstitial fibrosis.Kidney Int,1991,39(3):550
5 朱忠华,张春,邓安国.酶酚酸酯抑制大鼠肾间质纤维化的研究.中华肾脏病杂志,2002,18(4):295~297
6 左怡沁,马骥,顾勇,等.环加氧酶在梗阻性肾病大鼠肾脏的表 达 及 莫 比 可 的 作 用 机 制 研 究 . 中 华 肾 脏 病 杂志,2002,18(4):351~355
7 水华,陈德基,徐联芳,单侧输尿管大鼠肾小管间质纤维化的机制.武汉大学学报,2001,22(4):328~330
8 Perry M,Sutaria A Michael Jr Renal interstitial fibrosis is reduced in angiotensin II type la receptor-deficient mice. J AM SOC Nephrol,2001,12(2):317~325
9 曹骥翔,唐新玉,丁亮.黄芪、生脉注射液联合本那普利治疗 糖 尿 病 肾 病 32 例 疗 效 观 察 . 湖 南 中 医 药 导报,2002,8(10):596~597
10 牟娜,张庆怡,倪兆慧等.黄芪对高糖作用下肾间质成纤维细 胞 表 达 HGF 的 影 响 . 中 国 中 西 医 结 合 肾 病 杂志,2002,3(l):72
11 倪青.中医治疗慢性肾功能衰竭的思路与方法.辽宁中医杂志,2001,28(12):712~713
12 杜玉琴,李爱军,席瑞峰.黄芪注射液治疗慢性肾功能衰竭40 例临床分析.河北中医药,1999,20(2):121
13 宁英远,王俭勤,屈燧林. 大黄素对人肾成纤维细胞增殖的影响. 中国中西医结合杂志, 2000,20(2):105～107
14 刘冠贤,叶任高,谭志明,等.大黄素对狼疮性肾炎成纤维细胞 生 物 学 行 为 的 影 响 . 中 国 中 西 医 结 合 杂志,2000,20(3):196~198
15 远方,叶任高.叶任高治疗慢性肾功能衰竭经验集要.辽宁中医杂志,2001,28(6):336~337
16 魏向阳.大黄延缓慢性肾功能衰竭机制的研究进展.现代中医药,2003,3(1):58~59
17 彭佑铭,刘伏友,罗季安,等.丹参及生脉液对阿霉素所致大鼠肾小球硬化的实验治疗作用.湖南医科大学学报,1999,24(4):332
18 张萱,韩丽姝,徐晋等. 肾缺血再灌注损伤和丹参的保护作用的实验研究.解剖科学进展,1996,21(3) 269~272
19 沈寅初等. 丹参对实验性家兔肾缺血损伤防治作用的研究.遵义医学院学报,1986,9(4):26
20 沈寅初等. 丹参对实验性犬肾缺血损伤防治作用的研究.贵州医药,1988,12(1):32
21 张国强,叶任高,孔庆瑜等. 丹参对狼疮性肾炎成纤维 细胞增殖、凋亡及 c-myc 蛋白表达的的影响.中国中西医结合杂志,1997,17(8)473~475
22 胡顺金,恩因泽,方琦等. 黄芪及丹参注射液辅助治疗 慢性肾炎的疗效观察及机理探讨.安徽中医临床杂志, 1998,10(6) 350~352
23 乔保平,张亚伟,李道明等.丹参对大鼠环孢素 A 慢性 肾毒性的保护作用.河南医科大学学报,2000,35(4) 307~310
24 张步振等. 丹参对甘油引起的家兔急性肾功能衰竭的药效学研究.中国医药学报,1991,6(2):26
25 曾凡波,晏菊娇,万波,等.鳖甲煎丸药理学研究.中成药,2002,24(7):529～532
26 姚真敏,熊耀康,李剑平等. 鳖甲煎口服液抗肝纤维化作用的实验研究. 浙江中医学院学报,2000,2(1):58~59
27 都广礼,都广伍. 加减鳖甲煎汤对肝纤维化大鼠肝细胞心肌黄酶活性的影响. 辽宁中医学院学报,2000,2(3):82
28 高应斗,周雨凤.木通、茯苓、旋覆花的利尿作用.中华医学杂志,1955,( 10):963~966
29 陈春霞.茯苓多糖体的药理药化研究及其临床应用初探.中草药,1985,16(4):185~188
30 杜文摘译.利尿剂(猪苓汤、五苓散、柴苓汤)的作用机理.国外医学中医中药分册,1981,4(3):45
31 石纪才等.水蛭对缺血肾再灌注防治的实验性研究.天津医药,1992,20(3):166
32 Kuncio GS,Neilson EG, Haverly T.Mechanism of tubular interstitial fibrosis[J]. Kidney Int,1991,39:550~556
33 Massgaue J.Transfroming growth factor-β1family. Annui. Rev,1990,6:597
34 Wolf G, et al.Miner Electrolyte Metab,1998,24(2~ 3):174~180
35 TaiPale J,Shaharinen J,Hendman K,et al.Latent trans -forming.Growth factor-beta 1 and its binding protein are components of extracellular matrix microfibrils. Jhistochem,1996,44:875~889
36 Alvarez RJ,Sun MJ, Haverty TP, et al. Neilso eg: Biosynthetic and proliferative characteristics of tubulointerstitial fibroblasts probed with paracrine cytokines. Kidney Int 1992,41:14～23
37 Desmoulliere A, Geinoz A, Gabbiani F, et al.TGF-β1 induces a smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent cultured fibroblasts. J Cell Biol 1993,122:103～111
38 Badid C,Vincent M,Fouque D,et al.Myofibroblast:a prognostic marker and target cell in progressive renal disease.Ren Fail,2001,23:543~549
1 Gerhard A,Muller,Volke Sehettler,etal.Prevention of progression of renal fibrosis: How far are we.Kidney Int,1996,49(supply32):S75~S82
2 Nahas AME. Mechanisms of Progression and consequences of nephron reduction:inCamerons,Davison AM,Grunfeld JP , Kerr D, Ritz E(eds) Oxford TeXtbook of Clinical NePhrology, Oxford Univeisity Press,1992, (supply 34):1119~1226
3 RohleA,MullerGA,Wehrmann M,et al. Pathogenesis of chronic renal failure in the Primary glomerulo Pathies,renalvasculoPathies,and chronicinterstitial. Kidney Int,1996,49(supply 54):S2~S9
4 BholeA,GiseHV,Maekensen-Haen S,et al。 The obliteration of the Postglomerular capillaries and its influence upon the function of both glomeruli and tubuli. Klin Wbchensehr, 1981,59:1043~1051
5 Remuzzi G,Ruggeneti P Benigni A. Understanding the nature of renal disease progression. Kidney Int,1997,51:2~15
6 Bruijn JA,et al. J Lab Clin Med Biology of disease,atubular glomeruli and the structural basis for chronic renal failure,1998,111:140~156
7 Marcussen N. Biology of disease,atubular glomeruli and the struetural basis for chronic renal failure.LabInvest,1992,66:265~284
8 Wolf G, Ziyadeh FN, Stahl RA, et al. Angiotensin II stimulates expression of transforming growth factor beta receptor type II in cultured mouse proximal tubular cells. J Molmed, 1999, 77(5)556～564
9 Cheng SF, David H L. Gelatinase A (MMP-2) is necessary and sufficient for renal tubular cell epithelial mesenchymal transformation. Am J Pathol, 2003,162(12):1937～1949
10 王海燕.重视和研究小管间质损害在肾小球疾病发展和预后的作用.中华肾脏杂志,2000,16(1):5～6
11 李 竣,邹万卢,张波,等.病理性损伤因素对肾小管上皮细胞表型转化的影响.北京大学学报,2003,35(1):12～17
12 Klahr S, Morrissey JJ.The role of growth factors, cytokines, and vasoactive compounds in obstructive nephropathy. Semin Nephrol,1998,18(6):622～632
13 Alvarez RJ, Sun MJ, Haverty TP, et al. Neilso eg: biosynthetic and proliferative characteristics of tubulointerstitial fibroblasts probed with paracrine cytokines. Kidney Int, 1992;41:14～23
14 Desmoulliere A, Geinoz A, Gabbiani F, et al 。 TGF-β1induces a smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent cultured fibroblasts. J Cell Biol ,1993;122:103～111
15 Muchaneta-Kubara EC,el Nahas.Myofibroblast phenotypes expression in experimental renal scarring.Nephrol Dial Transplant,1997,12:904~915
16 Boukhalfa G,Desmouliere A,Rondeau E,et al.Relationship between alpha-smooth muscle actin expression and fibrotic changes in human kidney.Exp Nephrol,1996, 4(4):241~247
17 Ng YY,Huang TP,Yang WC,et al.Tubular epithelial -myofibroblast transdifferentiation in Dropressive tubulo -interstitial fibrosis in 5/6 nephrectomized rats.Kidney Int,1998,54(3):864~876
18 Goumenos D,Tsomi K,latrou C,et al.Myofibroblast and the progression of crescentic glomeulonephritis.Nephrol Dial Transplant,1998,13(7):1652~1661
19 乔保平,张亚伟,李道明等.丹参对大鼠环抱素 A 慢性肾毒性的保护用.河南医科大学学报,2000,35(4):307~310
20 丁一炜,李惊子,邹万忠等.黄芪当归合剂对肾病综合征鼠转 化 生 长 因 β1, 的 影 响 . 中 华 肾 脏 病 杂志,1998,14(4):229~232
21 李岩,陈香美,张颖等.中药海乐得对部分肾切除大鼠残余肾脏表达 PAI-1、TGF-βmRNA 的影响.中国病理生理杂志,1999,15(7):594~596
22 郭啸华,刘志红,戴春笋等.大黄酸抑制 TGF-β1,诱导的肾小管上皮细胞肥大及 ECM 产生.肾脏病透析与移植杂志,2001,10(2):101~105
23 刘冠贤,叶任高,谭志明等.大黄素对狼疮性肾炎成纤维细胞 生 物 学 行 为 的 影 响 . 中 国 中 西 医 结 合 杂志,2000,20(3):196~198
24 张国强,叶任高,孔庆瑜等.三七总苷诱导肾间质纤维化人肾成纤维细胞凋亡及其分子机理初探.中华肾脏病杂志,1998,14(2):93~95
25 屈隧林,方勤,陈高翔等.汉防己甲素、川芎嗪和苦杏仁苷对 人 肾 成 纤 维 细 胞 的 影 响 . 中 华 肾 脏 病 杂志,2000,16(3):186~189
26 王亚平,李伯样,王海泉等.红景天改善实验性大鼠肾间质纤维化的研究.中国中医药信息杂志,1998,5(11):21~22
27 孙万森,刘锐,乔成林等.绞股蓝总皂苷抗大鼠肾纤维化的实验研究.中国医药,1998,5(l):21~22
2 Kitamura K,Kangawa M,Kawamoto M,et al. Adrenome dullin:a novel hypotensive peptide isolated from human pheochromacytoma.Biochem BioPhys Res Commun, 1993,192:553~560
3 候凡凡.肾小管间质性疾病发病机理进展.国外医学内科分册,1991,18(8):361
4 Kuncio GS,Nilson EG,Haverty T.mechanisms of tubulionterstitial fibrosis.Kidney Int,1991,39(3):550
5 朱忠华,张春,邓安国.酶酚酸酯抑制大鼠肾间质纤维化的研究.中华肾脏病杂志,2002,18(4):295~297
6 左怡沁,马骥,顾勇,等.环加氧酶在梗阻性肾病大鼠肾脏的表 达 及 莫 比 可 的 作 用 机 制 研 究 . 中 华 肾 脏 病 杂志,2002,18(4):351~355
7 水华,陈德基,徐联芳,单侧输尿管大鼠肾小管间质纤维化的机制.武汉大学学报,2001,22(4):328~330
8 Perry M,Sutaria A Michael Jr Renal interstitial fibrosis is reduced in angiotensin II type la receptor-deficient mice. J AM SOC Nephrol,2001,12(2):317~325
9 曹骥翔,唐新玉,丁亮.黄芪、生脉注射液联合本那普利治疗 糖 尿 病 肾 病 32 例 疗 效 观 察 . 湖 南 中 医 药 导报,2002,8(10):596~597
10 牟娜,张庆怡,倪兆慧等.黄芪对高糖作用下肾间质成纤维细 胞 表 达 HGF 的 影 响 . 中 国 中 西 医 结 合 肾 病 杂志,2002,3(l):72
11 倪青.中医治疗慢性肾功能衰竭的思路与方法.辽宁中医杂志,2001,28(12):712~713
12 杜玉琴,李爱军,席瑞峰.黄芪注射液治疗慢性肾功能衰竭40 例临床分析.河北中医药,1999,20(2):121
13 宁英远,王俭勤,屈燧林. 大黄素对人肾成纤维细胞增殖的影响. 中国中西医结合杂志, 2000,20(2):105～107
14 刘冠贤,叶任高,谭志明,等.大黄素对狼疮性肾炎成纤维细胞 生 物 学 行 为 的 影 响 . 中 国 中 西 医 结 合 杂志,2000,20(3):196~198
15 远方,叶任高.叶任高治疗慢性肾功能衰竭经验集要.辽宁中医杂志,2001,28(6):336~337
16 魏向阳.大黄延缓慢性肾功能衰竭机制的研究进展.现代中医药,2003,3(1):58~59
17 彭佑铭,刘伏友,罗季安,等.丹参及生脉液对阿霉素所致大鼠肾小球硬化的实验治疗作用.湖南医科大学学报,1999,24(4):332
18 张萱,韩丽姝,徐晋等. 肾缺血再灌注损伤和丹参的保护作用的实验研究.解剖科学进展,1996,21(3) 269~272
19 沈寅初等. 丹参对实验性家兔肾缺血损伤防治作用的研究.遵义医学院学报,1986,9(4):26
20 沈寅初等. 丹参对实验性犬肾缺血损伤防治作用的研究.贵州医药,1988,12(1):32
21 张国强,叶任高,孔庆瑜等. 丹参对狼疮性肾炎成纤维 细胞增殖、凋亡及 c-myc 蛋白表达的的影响.中国中西医结合杂志,1997,17(8)473~475
22 胡顺金,恩因泽,方琦等. 黄芪及丹参注射液辅助治疗 慢性肾炎的疗效观察及机理探讨.安徽中医临床杂志, 1998,10(6) 350~352
23 乔保平,张亚伟,李道明等.丹参对大鼠环孢素 A 慢性 肾毒性的保护作用.河南医科大学学报,2000,35(4) 307~310
24 张步振等. 丹参对甘油引起的家兔急性肾功能衰竭的药效学研究.中国医药学报,1991,6(2):26
25 曾凡波,晏菊娇,万波,等.鳖甲煎丸药理学研究.中成药,2002,24(7):529～532
26 姚真敏,熊耀康,李剑平等. 鳖甲煎口服液抗肝纤维化作用的实验研究. 浙江中医学院学报,2000,2(1):58~59
27 都广礼,都广伍. 加减鳖甲煎汤对肝纤维化大鼠肝细胞心肌黄酶活性的影响. 辽宁中医学院学报,2000,2(3):82
28 高应斗,周雨凤.木通、茯苓、旋覆花的利尿作用.中华医学杂志,1955,( 10):963~966
29 陈春霞.茯苓多糖体的药理药化研究及其临床应用初探.中草药,1985,16(4):185~188
30 杜文摘译.利尿剂(猪苓汤、五苓散、柴苓汤)的作用机理.国外医学中医中药分册,1981,4(3):45
31 石纪才等.水蛭对缺血肾再灌注防治的实验性研究.天津医药,1992,20(3):166
32 Kuncio GS,Neilson EG, Haverly T.Mechanism of tubular interstitial fibrosis[J]. Kidney Int,1991,39:550~556
33 Massgaue J.Transfroming growth factor-β1family. Annui. Rev,1990,6:597
34 Wolf G, et al.Miner Electrolyte Metab,1998,24(2~ 3):174~180
35 TaiPale J,Shaharinen J,Hendman K,et al.Latent trans -forming.Growth factor-beta 1 and its binding protein are components of extracellular matrix microfibrils. Jhistochem,1996,44:875~889
36 Alvarez RJ,Sun MJ, Haverty TP, et al. Neilso eg: Biosynthetic and proliferative characteristics of tubulointerstitial fibroblasts probed with paracrine cytokines. Kidney Int 1992,41:14～23
37 Desmoulliere A, Geinoz A, Gabbiani F, et al.TGF-β1 induces a smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent cultured fibroblasts. J Cell Biol 1993,122:103～111
38 Badid C,Vincent M,Fouque D,et al.Myofibroblast:a prognostic marker and target cell in progressive renal disease.Ren Fail,2001,23:543~549
1 Gerhard A,Muller,Volke Sehettler,etal.Prevention of progression of renal fibrosis: How far are we.Kidney Int,1996,49(supply32):S75~S82
2 Nahas AME. Mechanisms of Progression and consequences of nephron reduction:inCamerons,Davison AM,Grunfeld JP , Kerr D, Ritz E(eds) Oxford TeXtbook of Clinical NePhrology, Oxford Univeisity Press,1992, (supply 34):1119~1226
3 RohleA,MullerGA,Wehrmann M,et al. Pathogenesis of chronic renal failure in the Primary glomerulo Pathies,renalvasculoPathies,and chronicinterstitial. Kidney Int,1996,49(supply 54):S2~S9
4 BholeA,GiseHV,Maekensen-Haen S,et al。 The obliteration of the Postglomerular capillaries and its influence upon the function of both glomeruli and tubuli. Klin Wbchensehr, 1981,59:1043~1051
5 Remuzzi G,Ruggeneti P Benigni A. Understanding the nature of renal disease progression. Kidney Int,1997,51:2~15
6 Bruijn JA,et al. J Lab Clin Med Biology of disease,atubular glomeruli and the structural basis for chronic renal failure,1998,111:140~156
7 Marcussen N. Biology of disease,atubular glomeruli and the struetural basis for chronic renal failure.LabInvest,1992,66:265~284
8 Wolf G, Ziyadeh FN, Stahl RA, et al. Angiotensin II stimulates expression of transforming growth factor beta receptor type II in cultured mouse proximal tubular cells. J Molmed, 1999, 77(5)556～564
9 Cheng SF, David H L. Gelatinase A (MMP-2) is necessary and sufficient for renal tubular cell epithelial mesenchymal transformation. Am J Pathol, 2003,162(12):1937～1949
10 王海燕.重视和研究小管间质损害在肾小球疾病发展和预后的作用.中华肾脏杂志,2000,16(1):5～6
11 李 竣,邹万卢,张波,等.病理性损伤因素对肾小管上皮细胞表型转化的影响.北京大学学报,2003,35(1):12～17
12 Klahr S, Morrissey JJ.The role of growth factors, cytokines, and vasoactive compounds in obstructive nephropathy. Semin Nephrol,1998,18(6):622～632
13 Alvarez RJ, Sun MJ, Haverty TP, et al. Neilso eg: biosynthetic and proliferative characteristics of tubulointerstitial fibroblasts probed with paracrine cytokines. Kidney Int, 1992;41:14～23
14 Desmoulliere A, Geinoz A, Gabbiani F, et al 。 TGF-β1induces a smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent cultured fibroblasts. J Cell Biol ,1993;122:103～111
15 Muchaneta-Kubara EC,el Nahas.Myofibroblast phenotypes expression in experimental renal scarring.Nephrol Dial Transplant,1997,12:904~915
16 Boukhalfa G,Desmouliere A,Rondeau E,et al.Relationship between alpha-smooth muscle actin expression and fibrotic changes in human kidney.Exp Nephrol,1996, 4(4):241~247
17 Ng YY,Huang TP,Yang WC,et al.Tubular epithelial -myofibroblast transdifferentiation in Dropressive tubulo -interstitial fibrosis in 5/6 nephrectomized rats.Kidney Int,1998,54(3):864~876
18 Goumenos D,Tsomi K,latrou C,et al.Myofibroblast and the progression of crescentic glomeulonephritis.Nephrol Dial Transplant,1998,13(7):1652~1661
19 乔保平,张亚伟,李道明等.丹参对大鼠环抱素 A 慢性肾毒性的保护用.河南医科大学学报,2000,35(4):307~310
20 丁一炜,李惊子,邹万忠等.黄芪当归合剂对肾病综合征鼠转 化 生 长 因 β1, 的 影 响 . 中 华 肾 脏 病 杂志,1998,14(4):229~232
21 李岩,陈香美,张颖等.中药海乐得对部分肾切除大鼠残余肾脏表达 PAI-1、TGF-βmRNA 的影响.中国病理生理杂志,1999,15(7):594~596
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