环维黄杨星D肾脏毒性机制的研究
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摘要
环维黄杨星D (Cyclovirobuxine D,简称CVB-D)是从黄杨科植物小叶黄杨(Buxus microphylla Sieb.et Zucc.Var.Sinica Rehd.et Wils)及其同属植物中提取得到的活性单体—生物碱,属孕甾烷衍生物,是黄杨宁片的主要成分。在临床上广泛用于心律失常、心绞痛、心肌缺血等治疗。近年来,对CVB-D的基础研究主要集中在其药理作用及其机制方面,毒理学研究报道较少。本实验室前期研究发现长期应用CVB-D能引起大鼠肾毒性,具有一定的剂量和时间依赖性关系,且病变在停药后不能彻底恢复。尽管如此,关于CVB-D肾脏毒性的机制目前尚无报道。基于此,我们将对CVB-D|肾脏损伤机制进行较为系统的研究,为临床应用提供合理依据。
本研究主要包括以下内容：
1.大鼠连续腹腔注射CVB-D两周后,检测各组大鼠尿液和血清中相关指标；检测肾皮质中生物膜和氧化相关指标；并做病理检查；此外基因芯片技术检测大鼠连续腹腔注射CVB-D 20 mg/kg 14天后与对照组大鼠肾脏基因表达的差异。结果显示,CVB-D各剂量组大鼠尿液中葡萄糖(Glu)和尿蛋白(TP)含量升高,血清中尿素氮(BUN)、肌酐(Cr)、Cl-含量升高并且降低大鼠肾皮质中Na+-K+-ATPase和Ca2+-Mg2+-ATPase活性,但是对脂质过氧化方面的指标没有显著影响。病理检测结果显示CVB-D可致肾小球毛细血管球细胞增生,肾小管上皮细胞有变性、坏死,肾间质有少量炎细胞浸润等现象；基因芯片检测结果显示,CVB-D对大鼠肾脏中凋亡、细胞周期、死亡受体等方面的相关通路和基因有所影响。以上结果显示CVB-D在较短时间内腹腔注射给药也会引起肾脏毒性,毒性机制与其引起肾小球、’肾小管损伤和抑制生物膜活性相关。
2.我们以HK-2细胞株为研究对象,在体外观察了CVB-D对肾小管上皮细胞的直接毒性。用比色法检测不同浓度CVB-D作用不同时间后HK-2细胞的活力变化、CVB-D对细胞LDH释放量的影响；倒置显微镜下观察CVB-D对细胞整体形态的影响；以及CVB-D对细胞周期和细胞内活性氧(ROS)水平的影响。结果发现,CVB-D可显著抑制HK-2细胞的增殖,促进细胞LDH的释放,且呈现时间和剂量依赖性,同时观察到细胞形态的变化如变圆、收缩,脱壁等变化。说明在实验设定的剂量和时间范围内,CVB-D对HK-2细胞具有明显的毒性作用。经CVB-D处理48h后,HK-2细胞出现S期阻滞,同时亚二倍体峰随着剂量的增加而增加；此外,CVB-D并未刺激HK-2细胞产生ROS。以上结果提示CVB-D对HK-2细胞的毒性可能与凋亡有关。
3.CVB-D处理HK-2细胞后,用透射电镜观察细胞形态学改变,并用Hoechst染色法观察细胞核形态变化；用Annexin V/PI双染法在流式细胞仪上检测CVB-D对HK-2细胞的凋亡诱导；用分光光度法检测CVB-D对HK-2细胞caspase-3活化程度的影响；基因芯片技术检测60μg/ml CVB-D作用HK-2细胞48h对其相关通路及基因表达变化的影响。结果显示,HK-2细胞出现了明显的染色质凝聚、边集等凋亡早期现象,细胞核染色同时显示出凋亡特征；AnnexinV/PI染色确证CVB-D处理后HK-2细胞出现早期凋亡,并且几乎没有晚期凋亡/坏死现象；CVB-D可活化HK-2细胞caspase-3;基因芯片结果显示,CVB-D可通过Caspase cascade, Fas和P53等凋亡通路影响HK-2细胞的凋亡,并且上调DIABLO, FADD和Gadd45a等促凋亡基因。上述现象说明,CVB-D对HK-2细胞的毒性作用主要是通过细胞凋亡的诱导而产生的,并且和caspase-3有关。此外,CVB-D可通过Caspase cascade, Fas和P53等凋亡通路影响HK-2细胞的凋亡。
4.观察CVB-D对Na+-K+-ATPase活性的影响,并用Western Blot方法检测CVB-D对Na+-K+-ATPaseα1和β1亚基蛋白表达的影响；此外,测定不同浓度CVB-D作用48h后对HK-2细胞线粒体膜电位和胞内Ca2+浓度的影响。结果显示,CVB-D对HK-2细胞Na+-K+-ATPase活性有显著抑制,具有浓度和时间的依赖性,并且显著下调了Na+-K+-ATPaseα1和Na+-K+-ATPaseβ1蛋白表达；CVB-D降低了HK-2细胞线粒体膜电位并且显著升高了胞内Ca2+浓度。以上实验结果显示,CVB-D通过下调HK-2细胞中Na+-K+-ATPaseα1和β1蛋白的表达从而抑制Na+-K+-ATPase的活性,导致细胞内Ca2+浓度升高,介导HK-2细胞凋亡
Cyclovirobuxine D (CVB-D), an active compound extracted from the Chinese medicinal herb Buxus microphylla Sieb. et Zucc. var. sinica Rehd. et Wils, is the active component of huangyangning tablets, which are widely used in China for the treatment of arrhythmias, angina pectoris, and myocardial ischemia. Numerous studies have examined the pharmacodynamic effects and mechanism of CVB-D, but few studies have examined its toxicity. We have found that long-term using CVB-D may cause nephrotoxicology, with the relationship between dose-dependent and time-dependent manners:and the nephrotoxicology cannot be recovered completely after injection. However, few studies have examined with its mechanism of kidney toxicity. In this study, we investigated the mechanism of CVB-D on nephrotoxicity to provide the reasonable reference for clinical practice.
1. After CVB-D was ip. given for two weeks, and we tested the relative indicators of urine and serum of rats. We also detected the biomembrane and oxidation indicators of renal cortex and observed pathological changes in renal tissue. Besides, we tested the differential gene expression between the CVB-D20 mg/kg group and control group using gene chip technology. Results showed compared with control group, Glu and TP in urine and BUN, Cr and Cr in serum of CVB-D group was increased; while the activity of Na+-K+-ATPase and Ca2+-Mg2+-ATPase was significant decreased in CVB-D group. CVB-D had few effects on oxidation indicators of renal cortex. Histopathology showed CVB-D caused capillary ball cell hyperplasia of glomerulus, denaturation and necrosis of renal tubular epithelial cells, and little inflammatory cell in renal interstitial. Gene chip showed CVB-D influenced the pathway about apoptosis, cell cycle, death receptor and so on. Those evidences above suggested that CVB-D caused the nephrotoxicity with ip.for shorter time, and the mechanism was associated with damage of glomerulus and tubles and inhibition of biomembrane activity.
2. We observed direct cytotoxic of CVB-D on HK-2cell in vitro by cell viability inhibition, LDH leakage, morphological changes, cell cycle changes and ROS generation. Results showed that CVB-D inhibited HK-2 cells viability and induced LDH leakage at a dose-and time-dependent manner. Treatment of CVB-D increased S phase and the SubGl phase appeared to be simultaneously. In addition, CVB-D had few effects on ROS generation. Those evidences above, especially the appearance of SubGl phase and turned round cells, have suggested that the cytotoxicities of CVB-D may related to apoptosis.
3. We observed cell morphology by electron microscope and cell nucleus morphology by Hoechst 33258 staining. Not only did we detect HK-2 cell apoptosis by annexinⅤ/PI staining combined with FCM analysis. We also detected the effects of CVB-D on caspase-3 activity by spectrophotometry and tested the differential gene expression between the CVB-D60μg/ml group and control group using gene chip technology. Results showed CVB-D group HK-2 cell exhibited chromatin margination, which was due to early apoptotic events. AnnexinV/PI staining confirmed the apoptosis was induced by CVB-D in HK-2 cell and without significant necrosis. In the meantime, CVB-D can activated caspase-3 and gene chip results showed it influenced HK-2 cell apoptosis according to caspase cascade, Fas and P53 pathway. All of above showed the CVB-D is able to induce apoptosis on HK-2 cell and caspase-3 is involved.
4. We observed the effects of CVB-D on Na+-K+-ATPase activity and tested expression of Na+-K+-ATPaseα1andβ1by western blotting. Besides, we tested the effects of CVB-D on mitochondrial membrane potential and intracellular Ca2+of HK-2 cell. Results showed that CVB-D can significant inhibited Na+-K+-ATPase activity and down regulated expression of Na+-K+-ATPaseα1 andβ1 at a dose-and time-dependent manner. In the meanwhile, CVB-D decreased mitochondrial membrane potential of HK-2 cell and increased intracellular Ca2+. All of above showed a significant decrease in Na+-K+-ATPase activity in cells treated with CVB-D, probably caused by the decrease in the expression of theα1 andβ1 subunit of Na+-K+-ATPase in HK-2 cells that may be associated with the CVB-D-induced apoptosis of HK-2 cells.
本研究主要包括以下内容：
1.大鼠连续腹腔注射CVB-D两周后,检测各组大鼠尿液和血清中相关指标；检测肾皮质中生物膜和氧化相关指标；并做病理检查；此外基因芯片技术检测大鼠连续腹腔注射CVB-D 20 mg/kg 14天后与对照组大鼠肾脏基因表达的差异。结果显示,CVB-D各剂量组大鼠尿液中葡萄糖(Glu)和尿蛋白(TP)含量升高,血清中尿素氮(BUN)、肌酐(Cr)、Cl-含量升高并且降低大鼠肾皮质中Na+-K+-ATPase和Ca2+-Mg2+-ATPase活性,但是对脂质过氧化方面的指标没有显著影响。病理检测结果显示CVB-D可致肾小球毛细血管球细胞增生,肾小管上皮细胞有变性、坏死,肾间质有少量炎细胞浸润等现象；基因芯片检测结果显示,CVB-D对大鼠肾脏中凋亡、细胞周期、死亡受体等方面的相关通路和基因有所影响。以上结果显示CVB-D在较短时间内腹腔注射给药也会引起肾脏毒性,毒性机制与其引起肾小球、’肾小管损伤和抑制生物膜活性相关。
2.我们以HK-2细胞株为研究对象,在体外观察了CVB-D对肾小管上皮细胞的直接毒性。用比色法检测不同浓度CVB-D作用不同时间后HK-2细胞的活力变化、CVB-D对细胞LDH释放量的影响；倒置显微镜下观察CVB-D对细胞整体形态的影响；以及CVB-D对细胞周期和细胞内活性氧(ROS)水平的影响。结果发现,CVB-D可显著抑制HK-2细胞的增殖,促进细胞LDH的释放,且呈现时间和剂量依赖性,同时观察到细胞形态的变化如变圆、收缩,脱壁等变化。说明在实验设定的剂量和时间范围内,CVB-D对HK-2细胞具有明显的毒性作用。经CVB-D处理48h后,HK-2细胞出现S期阻滞,同时亚二倍体峰随着剂量的增加而增加；此外,CVB-D并未刺激HK-2细胞产生ROS。以上结果提示CVB-D对HK-2细胞的毒性可能与凋亡有关。
3.CVB-D处理HK-2细胞后,用透射电镜观察细胞形态学改变,并用Hoechst染色法观察细胞核形态变化；用Annexin V/PI双染法在流式细胞仪上检测CVB-D对HK-2细胞的凋亡诱导；用分光光度法检测CVB-D对HK-2细胞caspase-3活化程度的影响；基因芯片技术检测60μg/ml CVB-D作用HK-2细胞48h对其相关通路及基因表达变化的影响。结果显示,HK-2细胞出现了明显的染色质凝聚、边集等凋亡早期现象,细胞核染色同时显示出凋亡特征；AnnexinV/PI染色确证CVB-D处理后HK-2细胞出现早期凋亡,并且几乎没有晚期凋亡/坏死现象；CVB-D可活化HK-2细胞caspase-3;基因芯片结果显示,CVB-D可通过Caspase cascade, Fas和P53等凋亡通路影响HK-2细胞的凋亡,并且上调DIABLO, FADD和Gadd45a等促凋亡基因。上述现象说明,CVB-D对HK-2细胞的毒性作用主要是通过细胞凋亡的诱导而产生的,并且和caspase-3有关。此外,CVB-D可通过Caspase cascade, Fas和P53等凋亡通路影响HK-2细胞的凋亡。
4.观察CVB-D对Na+-K+-ATPase活性的影响,并用Western Blot方法检测CVB-D对Na+-K+-ATPaseα1和β1亚基蛋白表达的影响；此外,测定不同浓度CVB-D作用48h后对HK-2细胞线粒体膜电位和胞内Ca2+浓度的影响。结果显示,CVB-D对HK-2细胞Na+-K+-ATPase活性有显著抑制,具有浓度和时间的依赖性,并且显著下调了Na+-K+-ATPaseα1和Na+-K+-ATPaseβ1蛋白表达；CVB-D降低了HK-2细胞线粒体膜电位并且显著升高了胞内Ca2+浓度。以上实验结果显示,CVB-D通过下调HK-2细胞中Na+-K+-ATPaseα1和β1蛋白的表达从而抑制Na+-K+-ATPase的活性,导致细胞内Ca2+浓度升高,介导HK-2细胞凋亡
Cyclovirobuxine D (CVB-D), an active compound extracted from the Chinese medicinal herb Buxus microphylla Sieb. et Zucc. var. sinica Rehd. et Wils, is the active component of huangyangning tablets, which are widely used in China for the treatment of arrhythmias, angina pectoris, and myocardial ischemia. Numerous studies have examined the pharmacodynamic effects and mechanism of CVB-D, but few studies have examined its toxicity. We have found that long-term using CVB-D may cause nephrotoxicology, with the relationship between dose-dependent and time-dependent manners:and the nephrotoxicology cannot be recovered completely after injection. However, few studies have examined with its mechanism of kidney toxicity. In this study, we investigated the mechanism of CVB-D on nephrotoxicity to provide the reasonable reference for clinical practice.
1. After CVB-D was ip. given for two weeks, and we tested the relative indicators of urine and serum of rats. We also detected the biomembrane and oxidation indicators of renal cortex and observed pathological changes in renal tissue. Besides, we tested the differential gene expression between the CVB-D20 mg/kg group and control group using gene chip technology. Results showed compared with control group, Glu and TP in urine and BUN, Cr and Cr in serum of CVB-D group was increased; while the activity of Na+-K+-ATPase and Ca2+-Mg2+-ATPase was significant decreased in CVB-D group. CVB-D had few effects on oxidation indicators of renal cortex. Histopathology showed CVB-D caused capillary ball cell hyperplasia of glomerulus, denaturation and necrosis of renal tubular epithelial cells, and little inflammatory cell in renal interstitial. Gene chip showed CVB-D influenced the pathway about apoptosis, cell cycle, death receptor and so on. Those evidences above suggested that CVB-D caused the nephrotoxicity with ip.for shorter time, and the mechanism was associated with damage of glomerulus and tubles and inhibition of biomembrane activity.
2. We observed direct cytotoxic of CVB-D on HK-2cell in vitro by cell viability inhibition, LDH leakage, morphological changes, cell cycle changes and ROS generation. Results showed that CVB-D inhibited HK-2 cells viability and induced LDH leakage at a dose-and time-dependent manner. Treatment of CVB-D increased S phase and the SubGl phase appeared to be simultaneously. In addition, CVB-D had few effects on ROS generation. Those evidences above, especially the appearance of SubGl phase and turned round cells, have suggested that the cytotoxicities of CVB-D may related to apoptosis.
3. We observed cell morphology by electron microscope and cell nucleus morphology by Hoechst 33258 staining. Not only did we detect HK-2 cell apoptosis by annexinⅤ/PI staining combined with FCM analysis. We also detected the effects of CVB-D on caspase-3 activity by spectrophotometry and tested the differential gene expression between the CVB-D60μg/ml group and control group using gene chip technology. Results showed CVB-D group HK-2 cell exhibited chromatin margination, which was due to early apoptotic events. AnnexinV/PI staining confirmed the apoptosis was induced by CVB-D in HK-2 cell and without significant necrosis. In the meantime, CVB-D can activated caspase-3 and gene chip results showed it influenced HK-2 cell apoptosis according to caspase cascade, Fas and P53 pathway. All of above showed the CVB-D is able to induce apoptosis on HK-2 cell and caspase-3 is involved.
4. We observed the effects of CVB-D on Na+-K+-ATPase activity and tested expression of Na+-K+-ATPaseα1andβ1by western blotting. Besides, we tested the effects of CVB-D on mitochondrial membrane potential and intracellular Ca2+of HK-2 cell. Results showed that CVB-D can significant inhibited Na+-K+-ATPase activity and down regulated expression of Na+-K+-ATPaseα1 andβ1 at a dose-and time-dependent manner. In the meanwhile, CVB-D decreased mitochondrial membrane potential of HK-2 cell and increased intracellular Ca2+. All of above showed a significant decrease in Na+-K+-ATPase activity in cells treated with CVB-D, probably caused by the decrease in the expression of theα1 andβ1 subunit of Na+-K+-ATPase in HK-2 cells that may be associated with the CVB-D-induced apoptosis of HK-2 cells.
引文
[1]梁秉文,张朝位.黄杨宁与心脑血管疾病[M].北京：蓝天出版社,2000：67
[2]梁涛,方泰惠,姚秀娟,等.环维黄杨星D约理研究进展[J].解放军约学学报,2001,17(1)：35-37
[3]周玖瑶.环维黄杨星D约理研究概况[J].广州中医约大学学报,2005,22(1)：70-74
[4]谭峰,顾卫,吴海科.环维黄杨星D的临床与实验研究近况[J].中国中医急症,2005,14(7)：683-684
[5]温业绍,周念辉,胡式冷,等.环维黄杨星D对心肌的止性肌力效应和细胞膜Na+-K+-ATPase活力的抑制作用[J].生理学报,1982,34(3)：295-303
[6]梁涛,方泰惠,姚秀娟,等.环维黄杨星D对培养的心肌细胞内游离钙浓度的影响[J].时珍国医国约,2001,12(5)：388-389
[7]梁涛,方泰惠,姚秀娟.环维黄杨星D升高心肌细胞内游离钙浓度实验研究[J].江苏药学与临床,2001,9(1)：9-10
[8]赖顺果,杜一鸣,章蕴毅.环维黄杨星D对心肌钙离子通道的作用[J].复旦学报(医学版),2005,32(2)：225-228
[9]Vacca G, Battaglia A, Brunellesehis S, et al. Hemodynamic effects of the intravenous Administration of cyclovirobuxine D in anesthetized pigs [J]. Life Sci.1997,61 (17):255-260
[10]Grossini E, Battaglia A, Brunellesehis S, et al. Coronary effeets of cyclovirobuxine D in anesthetized pigs and in isolated porcinecoronary arteries [J]. Life Sci,1999,65 (5):59-61
[11]汪永孝,郑云敏,范家驻,等.环维黄扬星D对正常和心肌缺血.大鼠血液流变学的影响[J].中国高血.压杂志.1995,3(2)：113-114
[12]Chen QW, Shan HL, Wang H, et al. Mechanisms of Cyclovirobuxine D on APD Prolongation in Rat Ventricular Myocytes[J]. Journal of Chinese Pharmaceutical Sciences.2003,12 (3):142-146
[13]方泰惠,伍必英,姚明辉,等.黄杨木碱1号对急性心肌缺血的实验研究[J].中草约通讯,1979(4)：33
[14]章蕴毅,王锦平,李佩芬,等.黄杨宁改进片增加动物冠脉流量和抗心律火常作用[J].中国临床约学杂志,2001,10(5)：310.
[15]叶寿山,下萍,倪光玉,等.注射用黄杨宁对心血管作用的研究[J].中约约理与临床2005,21(4)：16-20
[16]陈章强,胡申江,夏强,等.环维黄杨星D对大鼠心室乳头肌电生理作用[J].中国药学杂志,2004,39(7)：504-505
[17]陈庆文,单宏丽,孙宏丽,等.环维黄杨星D对分离大鼠心室肌细胞内Ca2+和L型钙电流的影响[J].约学学报,2004,39(7)：500-503
[18]陈章强,胡中江,史维雅,等.环维黄杨星D对大鼠心律失常双向作用的电生理研究[J].中国中西医结合杂志,2004,24(11)：1010-1013
[19]胡式冷,周念辉,范世藩.环维黄杨星D抗心律火常与诱发心律失常作用的实验分析[J].中国药理学报,1981,2(2)：101
[20]方泰惠,许惠,徐立,等.黄杨宁对急性实验性脑缺血的保护作用[J].中草约,1997,28(7)：413
[21]陶欣艺,沈祥春,方泰惠,等.环维黄杨星D对实验性脑缺血的保护作用[J].中约约理与临床,2003,19(5)：13-14
[22]谭峰,顾卫,吴海科,等.环维黄杨星D对高血压大鼠脑缺血损伤的保护作用[J].中国中西医结合急救杂志,2005,12(2)：97-100
[23]沈祥存,方泰惠,朱萱萱,等.环维黄杨星D对微循环的影响[J].中药约理与临床,2000,16(4)：15-16
[24]袁冬平,龙军,方泰惠.环维黄杨星D对PC12谷氨酸损伤的保护作用的实验研究[J].中西医结合心脑血.管病杂志,2004,2(1)：37-38
[25]李海涛,胡刚.环维黄杨星-D刘6-羟基多巴胺诱导的PC12细胞死亡率的影响[J].中国约科大学学报,2004,35(6)：562-564
[26]张惠勤,黄崇基.小叶黄杨约理研究初探[J].华夏医学,2005,18(3)：334-335
[27]程德培,吴华璞.黄杨宁对鼠整体氧耗量的动态观察[J].时珍国医国约,2002,13(10)：602-603.
[28]许立,梁涛,徐立,等.环维黄杨星D对大鼠肾脏毒性的初步观察[J].中药新药与临床药理,2007,18(2)：91-92
[29]梁晓雯,周静,黄艳等.环维黄杨星D对大鼠的肾脏毒性[J].中国约理学与毒理学,2010,24(6)：505-509
[30]杨霁云.约物性肾损伤[J].中国实用儿科杂志,2005,20(7)：387-390
[31]Baliga R, Ueda N, Walker PD, et al. Oxidant mechanisms in toxic acute renal failure[J]. Drug Metab Rev. 1999,31(4):971-975
[32]Lebeau C, Arlt VM, Schmeiser HH, et al. Aristolochic acid impedes endocytosis and induces DNA adducts in proximal tubule cells [J]. Kidney Int..2001,60(4):1332-1342
[33]Ryan MJ, Johnson G, Kirk J, HK-2:an immortalized proximal tubule epithelial cell line from normal adult human kidney [J]. Kidney Int.1994,45(1):48-57
[34]矫毓娟,刘江红.细胞凋亡的检测方法[J].中国神经免疫学和神经病学杂志,2004,11(1)：53-56
[35]Kerr JF, Wyllie AH, Currie AR. Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics [J]. Br J Cancer.1972,26(4):239-257
[36]Morishima N, Nakanishi K, Takenouchi H, et al. An endoplasmic reticulum stress-specific caspase cascade in apoptosis. Cytochrome c-independent activation of caspase-9 by caspase-12 [J]. Biol Chem, 2002,277 (37):34287-34294
[37]洪振亚Caspase非依赖性细胞程序性死亡[J].国际输血及血液学杂志,2006,29(2)：137-139
[38]姚君,王立生,高原Na+,K+-ATPase亚单位表达的研究进展[J].现代生物医学进展,2009,19(18)：3565-3568
[39]Shao Y, Ismail-Beigi F. Control of Na+,K+-ATPase beta 1-subunit expression:role of 3'-untranslated region [J]. Am J Physiol Cell Physiol,2004,286(3):580-585
[40]徐瑞成,张敏Na+,K+-ATP酶抑制引起的细胞凋亡和杂合性细胞死亡[J].细胞生物学杂志,2004,26：467-470
[41]Olej B, dos Santos NF, Leal L, et al. Ouabain induces apoptosis on PHA-activated lymphocytes [J]. Biosci Rep.1998,18(1):1-7
[42]Kurosawa M; Tani Y, Nishimura S. et al. Distinct PKC isozymes regulate bufalin-induced differentiation and apoptosis in human monocytic cells [J]. Am J Physiol Cell Physiol.2001,280(3):459-464
[43]Mann CL, Bortner CD, Jewell CM, et al. Glucocorticoid-induced plasma membrane depolarization during thymocyte apoptosis:association with cell shrinkage and degradation of the Na(+)/K(+)-adenosine triphosphatase [J]. Endocrinology.2001,142(12):5059-5068
[44]Wang H, Haas M, Liang M, et al. Ouabain assembles signaling cascades through the caveolar Na+,K+-ATPase [J]. J Biol Chem.2004,279(17):17250-17259
[45]Haas M, Wang H, Tian J, et al. Src-mediated inter-receptor cross-talk between the Na+,K+-ATPase and the epidermal growth factor receptor relays the signal from ouabain to mitogen-activated protein kinases [J]. J Biol Chem.2002,277(21):18694-18702
[46]王敏Na+,K+-ATP酶信号转导功能的分子机制[J].国际病理科学与临床杂志,2006,26(3)：228-231
[47]Yeh JY, Huang WJ, Kan SF, et al. Inhibitory effectsofdi-gitalison the proliferation of androgen dependent and in-dependent prostate cancer cells [J]. J Urol.2001,166(5):1937-1942
[48]KometianiP, Liu L, AskariA, et al. Digitalis-induced signaling by Na+,K+-ATPase in human breast cancer cells [J]. Mol Pharmacol.2005,67(3):929-936
[1]王林,陈大伟,曹瑾,等.铅镉联合暴露对大鼠肾脏功能损伤的研究[J].毒理学杂志,2008,22(5)：345-348
[2]刘湘群.44例糖尿病患者血尿酸与血.糖、尿素氮和肌酐的相关分析[J].当代医药卫生,2004,20：201-202
[3]金文达,宙义,陈锋,等.铅的肾脏毒性研究探讨[J].实用预防医学,2007,14(2)：597-599
[4]姚磊,盛鹏,朱德钟.肾综合征出血热病毒对体外人血管内皮细胞形态和ATP酶活性的影响[J].中国人兽共患病杂志,2001,17(4)：21-24
[5]李贞,高丽萍.氧化应激在顺铂肾毒性中的作用[J].广东医学,2010,31(19)：2600-2602
[6]于桂芬.活性氧清除剂对缺氧所致大鼠大脑皮层细胞产生脂质过氧化物的影响[J].中国病理生理杂志,1995,11(4)：378-381
[7]Rock MJ, Kincaid RL, Carstens GE. Effects of prenatal source and level of dietary selenium on passive immunity and thermometabolism of newborn lambs [J]. Small Ruminant Res.2001,40:129-138
[8]朱深银,周远大,杜冠华.二甘醇对大鼠肾脏的损伤作用[J].中国药理学与毒理学杂志,2009,23(2)：134-139
[9]梁涛,方泰惠,姚秀娟,等.环维黄杨星D对心肌细胞缺氧/复氧损伤的保护作用和对心肌细胞内游离钙浓度的影响[J].中国实验方剂学杂志,2009,15(12)：94-97
[10]Larisa L, Belyanskaya A, Algirdas Z. TRAIL-induced survival and proliferation of SCLC cells is mediated by ERK and dependenton TRAIL-R2/DR5 expression in the absence of Caspase-8[J]. Lang Cancer.2007,11(5):1-11
[11]Jin Z, El-Deiry WS. Overview of cell death signaling pathways [J]. Cancer Biol Ther.2005,4(2): 139-163
[12]张新生,左云飞,任双义tRNA转运与细胞周期检查点[J].生命的化学,2009,29(3)：335-337
[13]Kastan MB, Bartek J. Cell-cycle checkpoints and cancer [J]. Nature.2004,432(7015):316-323
[14]梁晓雯,周静,黄艳,等.环维黄杨星D对大鼠肾脏毒性的研究[J].中国药理学与毒理学杂志,2010,24(6)：505-509
[1]袁伯俊,廖明阳,李波.药物毒理学实验方法与技术[M].北京：化学工业出版社,2007,562
[2]Kim YM, de Vera ME, Watkins SC, et al. Nitric oxide protects cultured rat hepatocytes from tumor necrosis factor-alpha-induced apoptosis by inducing heat shock protein 70 expression. J Biol Chem. 1997,272(2):1402-1411
[3]Fadok VA, Voelker DR, Campbell PA, et al. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. The Journal of Immunology. 1992,148,2207-2216
[4]Sanchez Ⅰ, Dynlacht BD. New insights into cyclins, CDKs, and cell cycle control. Semin Cell Dev Biol. 2005,16(3):311-321
[5]任会荣,钱令嘉.线粒体通透性转换孔、细胞色素C与细胞凋亡[J].生命的化学,2002,22(4)：316-318
[6]郑荣梁,黄中洋.活性氧也许是端粒酶的调控者.科学通报,1999,24:2654-2658
[7]张惠勤,陈森洲,陈爱芳,等.环维黄杨星D对培养大鼠乳鼠心肌细胞缺氧/复氧损伤的保护作用.中国药理学通报,2007,23(10)：1338-1341
[1]Li J, Zhang L. Jiang Z, et al. Toxicities of aristolochic acid I and aristololactam I in cultured renal epithelial cells [J]. Toxicology in Vitro.2010,24,1092-1097
[2]Fadok VA, Voelker DR, Campbell PA, et al. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. [J]. J Immunol. 1992,148(7):2207-2216
[3]Daniel C, Duffield J. Brunner T, et al. Marti HP Matrix metalloproteinase inhibitors cause cell cycle arrest and apoptosis in glomerular mesangial cells [J]. J Pharmacol Exp Ther.2001.297,57-68
[4]Bassett DE Jr, Eisen MB, Boguski MS. Gene expression informatics--it's all in your mine [J]. Nat Genet. 1999.21(1 Suppl):51-55
[5]Medlin JF. Timely toxicology [J]. Environ Health Perspect.1999,107(5):A256-A258
[6]Vermes Ⅰ, Haanen C, Steffens-Nakken H, et al. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V [J]. Journal of Immunological Methods 1995,184,39-51
[7]Koopman G, Reutelingsperger CP, Kuijten GA, et al. Annexin Ⅴ for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis [J]. Blood 1994,84,1415-1420
[8]Schutte B. Nuydens R, Geerts H, et al. Annexin Ⅴ binding assay as a tool to measure apoptosis in differentiated neuronal cells [J]. Journal of Neuroscience Methods 1998,86,63-69
[9]汀立杰,叶良平,沈彤,等.三氯乙烯诱导人角质形成细胞凋亡中Caspase-3活力的研究[J].中国职业医学,2009,36(1)：7-10
[10]Fischer U, Janicke RU, Schulze-Osthoff K. Many cuts to ruin:a comprehensive update of caspase substrates [J]. Cell Death Differ.2003,10(1):76-100
[11]Thornberry NA, Lazebnik Y. Caspases:enemies within [J]. Science.1998,281(5381):1312-1316
[12]杨涛,杨琛懋,马璟,等.基因芯片技术研究威替米星及庆大霉素对HK-2细胞毒作用的机制[J].中国新药杂志,2007,16(3)：193-196
[13]Oltvai ZN, Milliman CI, Kovsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerats programmed all death [J]. Cell 1993,74:609-619
[14]Wang L. Du F, Wang X. TNF-alpha induces two distinct caspase-8 activation pathways [J]. Cell.2008, 133(4):693-703
[15]Bockbrader KM, Tan M. Sun Y. A small molecule Smac/DIABLO-mimic compound induces apoptosis and sensitizes TRAIL-and etoposide-induced apoptosis in breast cancer cells [J]. Oncogene,2005, 24(49):7381-7388
[16]Yamaka M, Yamagachi S, Sazuki T, et al. Apoptosis in rat cardiac myocytes induced by Fas ligand, priming for Fas-mediated apoptosis with doxotubicin [J]. J Mol Cell Cardiol.2000,32(6):881-889.
[17]于观贞,朱明华,陈颖,等.胰腺癌P53通路相关蛋白的表达及其临床意义[J].癌症.2005,24(11)：1398-1403
[18]M ChristineHollander, Albert J Fornace. Genomic instability, centrosome amplication. cell cycle checkpoints and Gadd45a [J]. Oncogene,2002,21(6):6228-6233
[19]池翠萍,李曙芳.137Csγ射线慢性照射刘CDKN1A基因表达的影响.辐射防护通讯,2010,30(5)：22-26
[20]AmourA, Slocombe PM, WebsterA, et al. TNF-alphconverting enzyme (TACE) is inhibited byTIMP-3[J]. FEBS Lett,1998,435(1):39-44
[21]周开梅,郭瑞珍,等.细胞周期蛋白在恶性肿瘤中的表达[J].医学综述,2010,16(4)：533-536
[22]Xu Q.Role of heat shock proteins in atherosclerosis [J]·Arterioscler Thromb Vasc Biol.2002,22 (10): 1547-1559
[23]Miramar MD, Costantini P, Ravagnan L. et al. NADH oxidase activity of mitochondrial apoptosis-inducing factor [J]. J Biol Chem.2001.276(19):16391-16398
[1]Scheiner-Bobis G.. The sodium pump. Its molecular properties and mechanics of ion transport[J]. Eur J Biochem.2002,269(10):2424-2433
[2]Dallaporta B, Hirsch T, Susin SA, et al. Potassium leakage during the apoptotic degradation phase [J]. J Immunol.1998,160(11):5605-5615
[3]Xiao AY, Wei L, Xia S, et al. Ionic mechanism of ouabain-induced concurrent apoptosis and necrosis in individual cultured cortical neurons[J]. J Neurosci.2002.22(4):1350-1362
[4]姚君,王立生,高原Na+,K+-ATPase亚单位表达的研究进展[J].现代生物医学进展.2009,19(18)：3565-3568
[5]Shoshani L,Contreras RG,Roldan ML,et al.The polarized expression of Na+,K+-ATPase in epithelia depends on the association between beta-subunits located in neighboring cells[J]. Mol Biol Cell.2005, 16(3):1071-1081
[6]温业绍.周念辉,胡式冷,等.环维黄杨星D对心肌的正性肌力效应和细胞膜Na+-K+-ATPase活力的抑制作用[J].生理学报,1982,34(3)：295-303
[7]Olson M, Kornbluth S. Mitochondria in apoptosis and human disease[J]. Curr Mol Med.2001,1(1): 91-122
[8]Wang H, Haas M, Liang M, et al. ouabain Assemble signaling cascades through the eaveolar Na+/K+-ATPase[J]. JBiolChem.2004,279(17):17250-17259
[9]McConkey DJ, Orrenius S. The role of calcium in the regulation of apoptosis[J]. Biochem Biophys Res Commun.1997,239(2):357-366
[10]张帅.崔隽,沈萍萍.细胞凋亡中的钙离子调控[J].细胞生物学杂志.2007,29：785-790
[11]Shpak B, Gofman Y, Shpak C, et al. Effects of purified endogenous inhibitor of the Na+ /Ca2+exchanger on ouabain-induced arrhythmias in the atria and ventricle strips of guinea pig[J].Eur J Pharmacol.2006,553(1-3):196-204
[12]陈小义,徐忠伟,王娜,等.哇巴因对血管内皮细胞ECV304细胞凋亡及胞内游离钙离子和活性氧浓度的影响[J].中国约理学与毒理学杂志.2010,24(4)：241-248
[13]唐海林.琦审校.钙离子信号与细胞凋亡关系的研究进展[J].南华大学学报,医学版,2006,34(5)：664-666
[14]Ashkenazi A, Dixit VM. Death receptors:signaling and modulation [J]. Science,1998,281(5381): 1305-1308
[15]Yeh JY, Huang WJ, Kan SF, et al. Inhibitory effectsofdi-gitalison the proliferation of androgen dependent and in-dependent prostate cancer cells [J]. J Urol.2001,166(5):1937-1942
[16]KometianiP, Liu L, AskariA, et al. Digitalis-induced signaling by Na+,K+-ATPase in human breast cancer cells [J]. Mol Pharmacol.2005,67(3):929-936
[1]梁秉文,张朝位.黄杨宁与心脑血管疾病[M].北京：蓝天出版社,2000.67
[2]梁涛,方泰惠,姚秀娟,等.环维黄杨星D约理研究进展[J].解放军药学学报,2001,17(1)：35-37
[3]周玖瑶.环维黄杨星D药理研究概况[J].广州中医约大学学报,2005,22(1)：70-74
[4]谭峰,顾卫,吴海科.环维黄杨星D的临床与实验研究近况[J].中国中医急症,2005,14(7)：683-684
[5]Liu XJ, Yao MH, Fang TH, et al. Some cardiovascular effects of cyclovirobuxine-D [J]. Acta Pharmacol Sin.1982,3(2):101-104
[6]Hu SL, Zhou NH, Fan SF. Experimental analysis of the anti-arrhythmic and arrhythmia-inducing actions of cyclovirbuxine D [J]. Acta Pharmacol Sin.1981.2(2):101-107
[7]邓兰,黄衡,徐鸣夏,等.环维黄杨星D的前药改造及生物活生研究[J].药学学报,2005,40(9)：820-824
[8]邓兰,黄衡,徐鸣夏,等.环维黄杨星D的结构改造及生物活性[J].药学学报,2004,39(6)：434-438
[9]陈爱芳,崔志清.黄杨宁的临床和实验研究近况[J].中华中医药学刊,25(4)：767-768
[10]柯仲成,桂双英,虞盛舟,等.黄杨宁的研究进展[J].甘肃中医学院学报,28(1)：71-73
[11]袁高辉,金培圣,邵华.黄杨宁对老年冠心病心功能不全者内源性洋地黄因子浓度影响的初步观察[J].中西医结合实用临床急救,1995,2(1)：6-7
[12]雷德培,朱瑞春,夏华珍,等.黄杨宁片治疗心律火常疗效观察[J].中成约,1992,14(2)：24-25
[13]牟乃洲,牛继华,高阳.黄杨宁治疗老年冠心病室性早搏52例[J].山东中医杂志,1996,15(7)：301-302
[14]蒋华民,李兆凤.黄杨宁片对高脂血症的临床与实验研究[J].中国中医约信息杂志,1999,6(6)：24-25
[15]严宗震,康庄.黄杨宁治疗低血压症45例疗效分析[J].中西医结合杂志,志,1990,10(6)：366
[16]刘如英.黄杨宁片的临床应用[J].临床内科杂志,1997,11(2)：112
[17]高华,卞如濂,陈修.约理实验方法学[M].北京：人民卫生出版社,1994：921
[18]康威,杜守颖,罗岸.黄杨宁缓释片的体外溶出度测定[J].北京中医约大学学报,2005,28(3)：65-67
[19]马凤余,毛幼桦,臧伟进,黄杨宁心血管药理作用[J].心脏杂志,2003,15(2)：169
[20]柯仲成,张辉,程子洋.黄杨宁分散片的制备工艺研究[J].黄山学院学报,2010,12(3)：40-42
[21]饶曼人,梁满达,刘广东,等.环常绿黄杨碱D对心肌梗死范围心肌代谢的影响[J].中草药,1984,15(4)：17
[22]方泰惠,伍必英,姚明辉,等.黄杨木碱1号对急性心肌缺血的实验研究[J].中草约通讯,1979,(4)：33
[23]陈俊,丁虹,童静,等.环维黄杨D对犬血流动力学及实验性心肌缺血的影响[J].中国药师,2005,8(8)：631-632
[24]刘锡明,姚明辉,方泰惠,等.环维黄杨星D的一些心血管作用[J].中国药理学报,1982,3(2)：101-103
[25]温业绍,周念辉,胡式冷,等.环维黄杨星D对心肌的正性肌力效应和细胞膜Na+-K+-ATPase活力的抑制作用[J].生理学报,1982,34(3)：295-297
[26]汪永孝,潭月华,盛宝恒.环维黄杨星D与哇巴因等约合用对离体心肌收缩力的影响[J].中国约学杂志,1994,29(1)：2-4
[27]梁涛,方泰惠,姚秀娟,等.环维黄杨星D对培养的心肌细胞内游离钙浓度的影响[J].时珍国医国约,2001,12(5)：388-390
[28]Vacca G, Battaglia A, Brunelleschi S, et al. Hemodynamiceffects of the intra-venous administration of cyclovirobu-xine Din anesthetized pigs [J]. Life Sci.1997,61(17):25-27
[29]汪永孝,郑云敏,范家骏,等.环维黄杨星D对正常和心肌缺血大鼠血液流变学的影响[J].中国高血压杂志,1995,3(2)：113-114
[30]梁秉文,张朝位.黄杨宁与心脑血管疾病[M].北京：蓝天出版社,2000,67-82
[31]章蕴毅,于锦平,李佩芬,等.黄杨宁改进片增加动物冠脉流量和抗心律失常作用[J].中国临床约学杂志,2001,10(5)：310-312
[32]陈章强,胡中江.环维黄杨星D对大鼠心律失常双向作用的电生理研究[J].中国中西医结合杂志,2004,24(11)：1010-1013
[33]Chen QW, ShanHL, WangH. et al. Mechanisms ofcycloviro-buxine D on APD prolongation of rat ventricularmyo-cytes [J]. Chin Pharm Sci.2003,12 (3):142-147
[34]汪永孝,郑云敏.环维黄杨星D抗心房纤颤的作用及其电生理机制[J].约学学报,1996,31(7)：481-486
[35]胡式冷,周念辉,范世藩.环维黄杨星D抗心律失常与诱发律火常作用的实验分析[J].中国约理 学报,1981,2(2)：101-104
[36]肖继皋,李选德,刘天培.环维黄杨星D对房室传导的影响—与利多卡的比较性研究[J].南京医学院学报,1984,4(1)：14-15
[37]沈祥春,方泰惠,朱萱萱,等.环维黄杨星D对微循环的影响[J].中药药理与临床,2000,16(4)：15-16
[38]Grossini E, Battaglia A, Brunelleschi S, et al. Coronary effects of cyclovirobuxine D in anesthetized pigs and in isolated porcinecoronary arteries [J]. Life Sci,1999,65(5):59-61
[39]袁冬平,龙军.环维黄杨星D对PC12谷氨酸损伤的保护作用的实验研究[J].中西医结合心脑血管病杂志,2004,2(1)：37-38
[40]张惠勤,黄崇基.小叶黄杨药理研究初探[J].华夏医学,2005,18(3)：334-335
[41]李海涛,胡刚CVB-D对6-羟基多巴胺诱导的PC12细胞死亡率的影响[J].中国药科大学学报,2004,35(6)：562-564
[42]程德培,吴华璞.黄杨宁对鼠整休氧耗量的动态观察[J].时珍国医国药,2002,13(10)：602-603
[43]余鹏,刘文英,高曙,等.环维黄杨星D在健康受试者体内的药动学研究[J].中国药学杂志,2008,43(3)：203-207
[44]Wen HM. Studies on the pharmacokinetics and disposition ofcyclovirbuxine D in rats and dogs[J]. China Pharmaceutical University,2003(5):44-46
[45]汀永孝,郑云敏.环维黄杨星D抗心房纤颤的作用及其电生理机制[J].药学学报,1996,31(7)：481-486
[46]许立,梁涛,徐立,等.环维黄杨星D对大鼠肾脏毒性的初步观察[J].中药新约与临床约理,2007,18(2)：91-92
[47]梁晓雯,周静,黄艳等.环维黄杨星D对大鼠的肾脏毒性[J].中国约理学与毒理学,2010,24(6)：505-509
[48]于民权,许立.环维黄杨星D对大鼠肾小管上皮细胞增殖的影响[J].江西中医药,39(311)：72-73
[2]梁涛,方泰惠,姚秀娟,等.环维黄杨星D约理研究进展[J].解放军约学学报,2001,17(1)：35-37
[3]周玖瑶.环维黄杨星D约理研究概况[J].广州中医约大学学报,2005,22(1)：70-74
[4]谭峰,顾卫,吴海科.环维黄杨星D的临床与实验研究近况[J].中国中医急症,2005,14(7)：683-684
[5]温业绍,周念辉,胡式冷,等.环维黄杨星D对心肌的止性肌力效应和细胞膜Na+-K+-ATPase活力的抑制作用[J].生理学报,1982,34(3)：295-303
[6]梁涛,方泰惠,姚秀娟,等.环维黄杨星D对培养的心肌细胞内游离钙浓度的影响[J].时珍国医国约,2001,12(5)：388-389
[7]梁涛,方泰惠,姚秀娟.环维黄杨星D升高心肌细胞内游离钙浓度实验研究[J].江苏药学与临床,2001,9(1)：9-10
[8]赖顺果,杜一鸣,章蕴毅.环维黄杨星D对心肌钙离子通道的作用[J].复旦学报(医学版),2005,32(2)：225-228
[9]Vacca G, Battaglia A, Brunellesehis S, et al. Hemodynamic effects of the intravenous Administration of cyclovirobuxine D in anesthetized pigs [J]. Life Sci.1997,61 (17):255-260
[10]Grossini E, Battaglia A, Brunellesehis S, et al. Coronary effeets of cyclovirobuxine D in anesthetized pigs and in isolated porcinecoronary arteries [J]. Life Sci,1999,65 (5):59-61
[11]汪永孝,郑云敏,范家驻,等.环维黄扬星D对正常和心肌缺血.大鼠血液流变学的影响[J].中国高血.压杂志.1995,3(2)：113-114
[12]Chen QW, Shan HL, Wang H, et al. Mechanisms of Cyclovirobuxine D on APD Prolongation in Rat Ventricular Myocytes[J]. Journal of Chinese Pharmaceutical Sciences.2003,12 (3):142-146
[13]方泰惠,伍必英,姚明辉,等.黄杨木碱1号对急性心肌缺血的实验研究[J].中草约通讯,1979(4)：33
[14]章蕴毅,王锦平,李佩芬,等.黄杨宁改进片增加动物冠脉流量和抗心律火常作用[J].中国临床约学杂志,2001,10(5)：310.
[15]叶寿山,下萍,倪光玉,等.注射用黄杨宁对心血管作用的研究[J].中约约理与临床2005,21(4)：16-20
[16]陈章强,胡申江,夏强,等.环维黄杨星D对大鼠心室乳头肌电生理作用[J].中国药学杂志,2004,39(7)：504-505
[17]陈庆文,单宏丽,孙宏丽,等.环维黄杨星D对分离大鼠心室肌细胞内Ca2+和L型钙电流的影响[J].约学学报,2004,39(7)：500-503
[18]陈章强,胡中江,史维雅,等.环维黄杨星D对大鼠心律失常双向作用的电生理研究[J].中国中西医结合杂志,2004,24(11)：1010-1013
[19]胡式冷,周念辉,范世藩.环维黄杨星D抗心律火常与诱发心律失常作用的实验分析[J].中国药理学报,1981,2(2)：101
[20]方泰惠,许惠,徐立,等.黄杨宁对急性实验性脑缺血的保护作用[J].中草约,1997,28(7)：413
[21]陶欣艺,沈祥春,方泰惠,等.环维黄杨星D对实验性脑缺血的保护作用[J].中约约理与临床,2003,19(5)：13-14
[22]谭峰,顾卫,吴海科,等.环维黄杨星D对高血压大鼠脑缺血损伤的保护作用[J].中国中西医结合急救杂志,2005,12(2)：97-100
[23]沈祥存,方泰惠,朱萱萱,等.环维黄杨星D对微循环的影响[J].中药约理与临床,2000,16(4)：15-16
[24]袁冬平,龙军,方泰惠.环维黄杨星D对PC12谷氨酸损伤的保护作用的实验研究[J].中西医结合心脑血.管病杂志,2004,2(1)：37-38
[25]李海涛,胡刚.环维黄杨星-D刘6-羟基多巴胺诱导的PC12细胞死亡率的影响[J].中国约科大学学报,2004,35(6)：562-564
[26]张惠勤,黄崇基.小叶黄杨约理研究初探[J].华夏医学,2005,18(3)：334-335
[27]程德培,吴华璞.黄杨宁对鼠整体氧耗量的动态观察[J].时珍国医国约,2002,13(10)：602-603.
[28]许立,梁涛,徐立,等.环维黄杨星D对大鼠肾脏毒性的初步观察[J].中药新药与临床药理,2007,18(2)：91-92
[29]梁晓雯,周静,黄艳等.环维黄杨星D对大鼠的肾脏毒性[J].中国约理学与毒理学,2010,24(6)：505-509
[30]杨霁云.约物性肾损伤[J].中国实用儿科杂志,2005,20(7)：387-390
[31]Baliga R, Ueda N, Walker PD, et al. Oxidant mechanisms in toxic acute renal failure[J]. Drug Metab Rev. 1999,31(4):971-975
[32]Lebeau C, Arlt VM, Schmeiser HH, et al. Aristolochic acid impedes endocytosis and induces DNA adducts in proximal tubule cells [J]. Kidney Int..2001,60(4):1332-1342
[33]Ryan MJ, Johnson G, Kirk J, HK-2:an immortalized proximal tubule epithelial cell line from normal adult human kidney [J]. Kidney Int.1994,45(1):48-57
[34]矫毓娟,刘江红.细胞凋亡的检测方法[J].中国神经免疫学和神经病学杂志,2004,11(1)：53-56
[35]Kerr JF, Wyllie AH, Currie AR. Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics [J]. Br J Cancer.1972,26(4):239-257
[36]Morishima N, Nakanishi K, Takenouchi H, et al. An endoplasmic reticulum stress-specific caspase cascade in apoptosis. Cytochrome c-independent activation of caspase-9 by caspase-12 [J]. Biol Chem, 2002,277 (37):34287-34294
[37]洪振亚Caspase非依赖性细胞程序性死亡[J].国际输血及血液学杂志,2006,29(2)：137-139
[38]姚君,王立生,高原Na+,K+-ATPase亚单位表达的研究进展[J].现代生物医学进展,2009,19(18)：3565-3568
[39]Shao Y, Ismail-Beigi F. Control of Na+,K+-ATPase beta 1-subunit expression:role of 3'-untranslated region [J]. Am J Physiol Cell Physiol,2004,286(3):580-585
[40]徐瑞成,张敏Na+,K+-ATP酶抑制引起的细胞凋亡和杂合性细胞死亡[J].细胞生物学杂志,2004,26：467-470
[41]Olej B, dos Santos NF, Leal L, et al. Ouabain induces apoptosis on PHA-activated lymphocytes [J]. Biosci Rep.1998,18(1):1-7
[42]Kurosawa M; Tani Y, Nishimura S. et al. Distinct PKC isozymes regulate bufalin-induced differentiation and apoptosis in human monocytic cells [J]. Am J Physiol Cell Physiol.2001,280(3):459-464
[43]Mann CL, Bortner CD, Jewell CM, et al. Glucocorticoid-induced plasma membrane depolarization during thymocyte apoptosis:association with cell shrinkage and degradation of the Na(+)/K(+)-adenosine triphosphatase [J]. Endocrinology.2001,142(12):5059-5068
[44]Wang H, Haas M, Liang M, et al. Ouabain assembles signaling cascades through the caveolar Na+,K+-ATPase [J]. J Biol Chem.2004,279(17):17250-17259
[45]Haas M, Wang H, Tian J, et al. Src-mediated inter-receptor cross-talk between the Na+,K+-ATPase and the epidermal growth factor receptor relays the signal from ouabain to mitogen-activated protein kinases [J]. J Biol Chem.2002,277(21):18694-18702
[46]王敏Na+,K+-ATP酶信号转导功能的分子机制[J].国际病理科学与临床杂志,2006,26(3)：228-231
[47]Yeh JY, Huang WJ, Kan SF, et al. Inhibitory effectsofdi-gitalison the proliferation of androgen dependent and in-dependent prostate cancer cells [J]. J Urol.2001,166(5):1937-1942
[48]KometianiP, Liu L, AskariA, et al. Digitalis-induced signaling by Na+,K+-ATPase in human breast cancer cells [J]. Mol Pharmacol.2005,67(3):929-936
[1]王林,陈大伟,曹瑾,等.铅镉联合暴露对大鼠肾脏功能损伤的研究[J].毒理学杂志,2008,22(5)：345-348
[2]刘湘群.44例糖尿病患者血尿酸与血.糖、尿素氮和肌酐的相关分析[J].当代医药卫生,2004,20：201-202
[3]金文达,宙义,陈锋,等.铅的肾脏毒性研究探讨[J].实用预防医学,2007,14(2)：597-599
[4]姚磊,盛鹏,朱德钟.肾综合征出血热病毒对体外人血管内皮细胞形态和ATP酶活性的影响[J].中国人兽共患病杂志,2001,17(4)：21-24
[5]李贞,高丽萍.氧化应激在顺铂肾毒性中的作用[J].广东医学,2010,31(19)：2600-2602
[6]于桂芬.活性氧清除剂对缺氧所致大鼠大脑皮层细胞产生脂质过氧化物的影响[J].中国病理生理杂志,1995,11(4)：378-381
[7]Rock MJ, Kincaid RL, Carstens GE. Effects of prenatal source and level of dietary selenium on passive immunity and thermometabolism of newborn lambs [J]. Small Ruminant Res.2001,40:129-138
[8]朱深银,周远大,杜冠华.二甘醇对大鼠肾脏的损伤作用[J].中国药理学与毒理学杂志,2009,23(2)：134-139
[9]梁涛,方泰惠,姚秀娟,等.环维黄杨星D对心肌细胞缺氧/复氧损伤的保护作用和对心肌细胞内游离钙浓度的影响[J].中国实验方剂学杂志,2009,15(12)：94-97
[10]Larisa L, Belyanskaya A, Algirdas Z. TRAIL-induced survival and proliferation of SCLC cells is mediated by ERK and dependenton TRAIL-R2/DR5 expression in the absence of Caspase-8[J]. Lang Cancer.2007,11(5):1-11
[11]Jin Z, El-Deiry WS. Overview of cell death signaling pathways [J]. Cancer Biol Ther.2005,4(2): 139-163
[12]张新生,左云飞,任双义tRNA转运与细胞周期检查点[J].生命的化学,2009,29(3)：335-337
[13]Kastan MB, Bartek J. Cell-cycle checkpoints and cancer [J]. Nature.2004,432(7015):316-323
[14]梁晓雯,周静,黄艳,等.环维黄杨星D对大鼠肾脏毒性的研究[J].中国药理学与毒理学杂志,2010,24(6)：505-509
[1]袁伯俊,廖明阳,李波.药物毒理学实验方法与技术[M].北京：化学工业出版社,2007,562
[2]Kim YM, de Vera ME, Watkins SC, et al. Nitric oxide protects cultured rat hepatocytes from tumor necrosis factor-alpha-induced apoptosis by inducing heat shock protein 70 expression. J Biol Chem. 1997,272(2):1402-1411
[3]Fadok VA, Voelker DR, Campbell PA, et al. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. The Journal of Immunology. 1992,148,2207-2216
[4]Sanchez Ⅰ, Dynlacht BD. New insights into cyclins, CDKs, and cell cycle control. Semin Cell Dev Biol. 2005,16(3):311-321
[5]任会荣,钱令嘉.线粒体通透性转换孔、细胞色素C与细胞凋亡[J].生命的化学,2002,22(4)：316-318
[6]郑荣梁,黄中洋.活性氧也许是端粒酶的调控者.科学通报,1999,24:2654-2658
[7]张惠勤,陈森洲,陈爱芳,等.环维黄杨星D对培养大鼠乳鼠心肌细胞缺氧/复氧损伤的保护作用.中国药理学通报,2007,23(10)：1338-1341
[1]Li J, Zhang L. Jiang Z, et al. Toxicities of aristolochic acid I and aristololactam I in cultured renal epithelial cells [J]. Toxicology in Vitro.2010,24,1092-1097
[2]Fadok VA, Voelker DR, Campbell PA, et al. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. [J]. J Immunol. 1992,148(7):2207-2216
[3]Daniel C, Duffield J. Brunner T, et al. Marti HP Matrix metalloproteinase inhibitors cause cell cycle arrest and apoptosis in glomerular mesangial cells [J]. J Pharmacol Exp Ther.2001.297,57-68
[4]Bassett DE Jr, Eisen MB, Boguski MS. Gene expression informatics--it's all in your mine [J]. Nat Genet. 1999.21(1 Suppl):51-55
[5]Medlin JF. Timely toxicology [J]. Environ Health Perspect.1999,107(5):A256-A258
[6]Vermes Ⅰ, Haanen C, Steffens-Nakken H, et al. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V [J]. Journal of Immunological Methods 1995,184,39-51
[7]Koopman G, Reutelingsperger CP, Kuijten GA, et al. Annexin Ⅴ for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis [J]. Blood 1994,84,1415-1420
[8]Schutte B. Nuydens R, Geerts H, et al. Annexin Ⅴ binding assay as a tool to measure apoptosis in differentiated neuronal cells [J]. Journal of Neuroscience Methods 1998,86,63-69
[9]汀立杰,叶良平,沈彤,等.三氯乙烯诱导人角质形成细胞凋亡中Caspase-3活力的研究[J].中国职业医学,2009,36(1)：7-10
[10]Fischer U, Janicke RU, Schulze-Osthoff K. Many cuts to ruin:a comprehensive update of caspase substrates [J]. Cell Death Differ.2003,10(1):76-100
[11]Thornberry NA, Lazebnik Y. Caspases:enemies within [J]. Science.1998,281(5381):1312-1316
[12]杨涛,杨琛懋,马璟,等.基因芯片技术研究威替米星及庆大霉素对HK-2细胞毒作用的机制[J].中国新药杂志,2007,16(3)：193-196
[13]Oltvai ZN, Milliman CI, Kovsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerats programmed all death [J]. Cell 1993,74:609-619
[14]Wang L. Du F, Wang X. TNF-alpha induces two distinct caspase-8 activation pathways [J]. Cell.2008, 133(4):693-703
[15]Bockbrader KM, Tan M. Sun Y. A small molecule Smac/DIABLO-mimic compound induces apoptosis and sensitizes TRAIL-and etoposide-induced apoptosis in breast cancer cells [J]. Oncogene,2005, 24(49):7381-7388
[16]Yamaka M, Yamagachi S, Sazuki T, et al. Apoptosis in rat cardiac myocytes induced by Fas ligand, priming for Fas-mediated apoptosis with doxotubicin [J]. J Mol Cell Cardiol.2000,32(6):881-889.
[17]于观贞,朱明华,陈颖,等.胰腺癌P53通路相关蛋白的表达及其临床意义[J].癌症.2005,24(11)：1398-1403
[18]M ChristineHollander, Albert J Fornace. Genomic instability, centrosome amplication. cell cycle checkpoints and Gadd45a [J]. Oncogene,2002,21(6):6228-6233
[19]池翠萍,李曙芳.137Csγ射线慢性照射刘CDKN1A基因表达的影响.辐射防护通讯,2010,30(5)：22-26
[20]AmourA, Slocombe PM, WebsterA, et al. TNF-alphconverting enzyme (TACE) is inhibited byTIMP-3[J]. FEBS Lett,1998,435(1):39-44
[21]周开梅,郭瑞珍,等.细胞周期蛋白在恶性肿瘤中的表达[J].医学综述,2010,16(4)：533-536
[22]Xu Q.Role of heat shock proteins in atherosclerosis [J]·Arterioscler Thromb Vasc Biol.2002,22 (10): 1547-1559
[23]Miramar MD, Costantini P, Ravagnan L. et al. NADH oxidase activity of mitochondrial apoptosis-inducing factor [J]. J Biol Chem.2001.276(19):16391-16398
[1]Scheiner-Bobis G.. The sodium pump. Its molecular properties and mechanics of ion transport[J]. Eur J Biochem.2002,269(10):2424-2433
[2]Dallaporta B, Hirsch T, Susin SA, et al. Potassium leakage during the apoptotic degradation phase [J]. J Immunol.1998,160(11):5605-5615
[3]Xiao AY, Wei L, Xia S, et al. Ionic mechanism of ouabain-induced concurrent apoptosis and necrosis in individual cultured cortical neurons[J]. J Neurosci.2002.22(4):1350-1362
[4]姚君,王立生,高原Na+,K+-ATPase亚单位表达的研究进展[J].现代生物医学进展.2009,19(18)：3565-3568
[5]Shoshani L,Contreras RG,Roldan ML,et al.The polarized expression of Na+,K+-ATPase in epithelia depends on the association between beta-subunits located in neighboring cells[J]. Mol Biol Cell.2005, 16(3):1071-1081
[6]温业绍.周念辉,胡式冷,等.环维黄杨星D对心肌的正性肌力效应和细胞膜Na+-K+-ATPase活力的抑制作用[J].生理学报,1982,34(3)：295-303
[7]Olson M, Kornbluth S. Mitochondria in apoptosis and human disease[J]. Curr Mol Med.2001,1(1): 91-122
[8]Wang H, Haas M, Liang M, et al. ouabain Assemble signaling cascades through the eaveolar Na+/K+-ATPase[J]. JBiolChem.2004,279(17):17250-17259
[9]McConkey DJ, Orrenius S. The role of calcium in the regulation of apoptosis[J]. Biochem Biophys Res Commun.1997,239(2):357-366
[10]张帅.崔隽,沈萍萍.细胞凋亡中的钙离子调控[J].细胞生物学杂志.2007,29：785-790
[11]Shpak B, Gofman Y, Shpak C, et al. Effects of purified endogenous inhibitor of the Na+ /Ca2+exchanger on ouabain-induced arrhythmias in the atria and ventricle strips of guinea pig[J].Eur J Pharmacol.2006,553(1-3):196-204
[12]陈小义,徐忠伟,王娜,等.哇巴因对血管内皮细胞ECV304细胞凋亡及胞内游离钙离子和活性氧浓度的影响[J].中国约理学与毒理学杂志.2010,24(4)：241-248
[13]唐海林.琦审校.钙离子信号与细胞凋亡关系的研究进展[J].南华大学学报,医学版,2006,34(5)：664-666
[14]Ashkenazi A, Dixit VM. Death receptors:signaling and modulation [J]. Science,1998,281(5381): 1305-1308
[15]Yeh JY, Huang WJ, Kan SF, et al. Inhibitory effectsofdi-gitalison the proliferation of androgen dependent and in-dependent prostate cancer cells [J]. J Urol.2001,166(5):1937-1942
[16]KometianiP, Liu L, AskariA, et al. Digitalis-induced signaling by Na+,K+-ATPase in human breast cancer cells [J]. Mol Pharmacol.2005,67(3):929-936
[1]梁秉文,张朝位.黄杨宁与心脑血管疾病[M].北京：蓝天出版社,2000.67
[2]梁涛,方泰惠,姚秀娟,等.环维黄杨星D约理研究进展[J].解放军药学学报,2001,17(1)：35-37
[3]周玖瑶.环维黄杨星D药理研究概况[J].广州中医约大学学报,2005,22(1)：70-74
[4]谭峰,顾卫,吴海科.环维黄杨星D的临床与实验研究近况[J].中国中医急症,2005,14(7)：683-684
[5]Liu XJ, Yao MH, Fang TH, et al. Some cardiovascular effects of cyclovirobuxine-D [J]. Acta Pharmacol Sin.1982,3(2):101-104
[6]Hu SL, Zhou NH, Fan SF. Experimental analysis of the anti-arrhythmic and arrhythmia-inducing actions of cyclovirbuxine D [J]. Acta Pharmacol Sin.1981.2(2):101-107
[7]邓兰,黄衡,徐鸣夏,等.环维黄杨星D的前药改造及生物活生研究[J].药学学报,2005,40(9)：820-824
[8]邓兰,黄衡,徐鸣夏,等.环维黄杨星D的结构改造及生物活性[J].药学学报,2004,39(6)：434-438
[9]陈爱芳,崔志清.黄杨宁的临床和实验研究近况[J].中华中医药学刊,25(4)：767-768
[10]柯仲成,桂双英,虞盛舟,等.黄杨宁的研究进展[J].甘肃中医学院学报,28(1)：71-73
[11]袁高辉,金培圣,邵华.黄杨宁对老年冠心病心功能不全者内源性洋地黄因子浓度影响的初步观察[J].中西医结合实用临床急救,1995,2(1)：6-7
[12]雷德培,朱瑞春,夏华珍,等.黄杨宁片治疗心律火常疗效观察[J].中成约,1992,14(2)：24-25
[13]牟乃洲,牛继华,高阳.黄杨宁治疗老年冠心病室性早搏52例[J].山东中医杂志,1996,15(7)：301-302
[14]蒋华民,李兆凤.黄杨宁片对高脂血症的临床与实验研究[J].中国中医约信息杂志,1999,6(6)：24-25
[15]严宗震,康庄.黄杨宁治疗低血压症45例疗效分析[J].中西医结合杂志,志,1990,10(6)：366
[16]刘如英.黄杨宁片的临床应用[J].临床内科杂志,1997,11(2)：112
[17]高华,卞如濂,陈修.约理实验方法学[M].北京：人民卫生出版社,1994：921
[18]康威,杜守颖,罗岸.黄杨宁缓释片的体外溶出度测定[J].北京中医约大学学报,2005,28(3)：65-67
[19]马凤余,毛幼桦,臧伟进,黄杨宁心血管药理作用[J].心脏杂志,2003,15(2)：169
[20]柯仲成,张辉,程子洋.黄杨宁分散片的制备工艺研究[J].黄山学院学报,2010,12(3)：40-42
[21]饶曼人,梁满达,刘广东,等.环常绿黄杨碱D对心肌梗死范围心肌代谢的影响[J].中草药,1984,15(4)：17
[22]方泰惠,伍必英,姚明辉,等.黄杨木碱1号对急性心肌缺血的实验研究[J].中草约通讯,1979,(4)：33
[23]陈俊,丁虹,童静,等.环维黄杨D对犬血流动力学及实验性心肌缺血的影响[J].中国药师,2005,8(8)：631-632
[24]刘锡明,姚明辉,方泰惠,等.环维黄杨星D的一些心血管作用[J].中国药理学报,1982,3(2)：101-103
[25]温业绍,周念辉,胡式冷,等.环维黄杨星D对心肌的正性肌力效应和细胞膜Na+-K+-ATPase活力的抑制作用[J].生理学报,1982,34(3)：295-297
[26]汪永孝,潭月华,盛宝恒.环维黄杨星D与哇巴因等约合用对离体心肌收缩力的影响[J].中国约学杂志,1994,29(1)：2-4
[27]梁涛,方泰惠,姚秀娟,等.环维黄杨星D对培养的心肌细胞内游离钙浓度的影响[J].时珍国医国约,2001,12(5)：388-390
[28]Vacca G, Battaglia A, Brunelleschi S, et al. Hemodynamiceffects of the intra-venous administration of cyclovirobu-xine Din anesthetized pigs [J]. Life Sci.1997,61(17):25-27
[29]汪永孝,郑云敏,范家骏,等.环维黄杨星D对正常和心肌缺血大鼠血液流变学的影响[J].中国高血压杂志,1995,3(2)：113-114
[30]梁秉文,张朝位.黄杨宁与心脑血管疾病[M].北京：蓝天出版社,2000,67-82
[31]章蕴毅,于锦平,李佩芬,等.黄杨宁改进片增加动物冠脉流量和抗心律失常作用[J].中国临床约学杂志,2001,10(5)：310-312
[32]陈章强,胡中江.环维黄杨星D对大鼠心律失常双向作用的电生理研究[J].中国中西医结合杂志,2004,24(11)：1010-1013
[33]Chen QW, ShanHL, WangH. et al. Mechanisms ofcycloviro-buxine D on APD prolongation of rat ventricularmyo-cytes [J]. Chin Pharm Sci.2003,12 (3):142-147
[34]汪永孝,郑云敏.环维黄杨星D抗心房纤颤的作用及其电生理机制[J].约学学报,1996,31(7)：481-486
[35]胡式冷,周念辉,范世藩.环维黄杨星D抗心律失常与诱发律火常作用的实验分析[J].中国约理 学报,1981,2(2)：101-104
[36]肖继皋,李选德,刘天培.环维黄杨星D对房室传导的影响—与利多卡的比较性研究[J].南京医学院学报,1984,4(1)：14-15
[37]沈祥春,方泰惠,朱萱萱,等.环维黄杨星D对微循环的影响[J].中药药理与临床,2000,16(4)：15-16
[38]Grossini E, Battaglia A, Brunelleschi S, et al. Coronary effects of cyclovirobuxine D in anesthetized pigs and in isolated porcinecoronary arteries [J]. Life Sci,1999,65(5):59-61
[39]袁冬平,龙军.环维黄杨星D对PC12谷氨酸损伤的保护作用的实验研究[J].中西医结合心脑血管病杂志,2004,2(1)：37-38
[40]张惠勤,黄崇基.小叶黄杨药理研究初探[J].华夏医学,2005,18(3)：334-335
[41]李海涛,胡刚CVB-D对6-羟基多巴胺诱导的PC12细胞死亡率的影响[J].中国药科大学学报,2004,35(6)：562-564
[42]程德培,吴华璞.黄杨宁对鼠整休氧耗量的动态观察[J].时珍国医国药,2002,13(10)：602-603
[43]余鹏,刘文英,高曙,等.环维黄杨星D在健康受试者体内的药动学研究[J].中国药学杂志,2008,43(3)：203-207
[44]Wen HM. Studies on the pharmacokinetics and disposition ofcyclovirbuxine D in rats and dogs[J]. China Pharmaceutical University,2003(5):44-46
[45]汀永孝,郑云敏.环维黄杨星D抗心房纤颤的作用及其电生理机制[J].药学学报,1996,31(7)：481-486
[46]许立,梁涛,徐立,等.环维黄杨星D对大鼠肾脏毒性的初步观察[J].中药新约与临床约理,2007,18(2)：91-92
[47]梁晓雯,周静,黄艳等.环维黄杨星D对大鼠的肾脏毒性[J].中国约理学与毒理学,2010,24(6)：505-509
[48]于民权,许立.环维黄杨星D对大鼠肾小管上皮细胞增殖的影响[J].江西中医药,39(311)：72-73