川芎嗪对缺血/再灌注损伤大鼠肾脏细胞凋亡及Bcl-2和Bax表达的影响

Effects of ligustrazine on cell apoptosis and expressions of Bcl-2 and Bax in rat kidneys with ischemia-reperfusion injury

目的:观察川芎嗪对急性缺血/再灌注(I/R)损伤大鼠肾脏细胞凋亡及凋亡相关蛋白的影响,探讨川芎嗪对急性肾I/R损伤保护作用的可能机制.方法:将40只Wistar大鼠夹闭双侧肾动脉45min再灌注24h,制备成急性肾I/R损伤动物模型,随机分为假手术对照组、I/R组、川芎嗪治疗组和川芎嗪预防组,采用原位末端标记法检测细胞凋亡指数,免疫组化法测定Bcl-2,Bax表达,电镜观察肾组织细胞超微结构.结果:I/R组较假手术对照组肾小管细胞凋亡指数明显增多(28.8±4.6vs1.9±0.5,P<0.01),Bax表达显著增强(162.6±17.1vs182.7±12.8,P<0.01),Bcl-2/Bax显著降低(1.1±0.1vs1.0±0.1,P<0.01);川芎嗪预防组较I/R组肾小管凋亡细胞数明显减少(13.6±2.9vs28.8±4.6,P<0.05),Bax表达明显减弱(179.1±12.7vs162.6±17.1,P<0.05),Bcl-2表达明显增强(166.6±15.1vs178.7±13.0,P<0.05),Bcl-2/Bax显著增高(0.9±0.1vs1.1±0.1,P<0.05).结论:川芎嗪对急性肾I/R损伤具有保护作用,其作用机制可能是通过调节凋亡相关基因Bcl-2和Bax介导的I/R损伤肾脏细胞凋亡而实现.

AIM： To investigate the effects of ligustrazine on cell apoptosis and expressions of Bcl-2 and Bax in rat kidneys with acute ischemia-reperfusion （I/R） injury, and study the possible mechanism of renal protection affected by ligustrazine against acute I/R injury. METHODS： Forty Wistar rats of either sex were randomly divided into 4 groups, which were sham operation （ n = 10 ）, I/R model （ n = 10 ）, intraperitoneal ligustrazine 32 mg/kg every 6 h after reperfusion for 30 min （n = 10） and intraperitoneal ligustrazine 32 mg/kg at 45 min, 8, 16, 24 h before ischemia （n = 10 ）. The models were established through clamping bilateral renal arteries for 45 min and then reperfusing for 24 h. Cell apoptotic index was examined by means of terminal deoxynucleotidyl transferase-mediated d-UTP nick end labeling （TUNEL）. The expressions of Bcl-2 and Bax protein were measured by immunohistochemical technique and the kidney uhrastrucutre was observed by electron microscope. RESULTS： Compared with those in the control group, the cell apoptotic index （ 1.9 ± 0. 5 vs 28.8 ± 4.6, P 〈 0.01 ） and the expression of Bax （ 162.6 ± 17.1 vs 182.7 ± 12.8, P 〈0.01 ） were significantly increased, and the Bcl-2/Bax ratio （ 1.1 ±0. 1 vs 1.0 ±0. 1, P 〈0.01 ） was decreased significantly in I/R model group. By ligustrazine pretreatment, the apoptotic index（28.8 ±4.6 vs 13.6 ±2.9, P 〈 0.05 ） and the expression of Bax were decreased significantly （ 179.1 ± 12.7 vs 162.6 ± 17. 1, P 〈 0. 05 ）, the expression of Bcl-2 （ 166.6 ± 15.1 vs 178.7 ± 13.0, P 〈 0.05） and the Bcl-2/Bax ratio（0.9 ±0.1 vs 1.1 ±0.1, P 〈0.05） were increased significantly. CONCLUSION： Ligustrazine can protect kidney from acute I/R injury, partly through inhibition of cell apoptosis by regulating the expressions of apoptosis-related genes Bcl-2 and Bax.