制大黄-川芎药对对造影剂肾病大鼠肾小管上皮细胞凋亡的影响

Effects of Zhidahuang-Chuanxiong Drug Pair on Tubular Epithelial Cell Apoptosis in Contrast-induced Nephropathy Rats

目的:探讨制大黄-川芎药对对造影剂肾病(CIN)大鼠肾小管上皮细胞凋亡的影响及机制。方法:将32只雄性SD大鼠分为正常组(A组)、模型组(B组)、药对高剂量组(C组)、药对低剂量组(D组)。C、D组于造模前7天每日灌胃药对水煎液,灌胃量分别为成人标准体重(60kg)常规用量50、20倍。造模后24h处死动物,测定血清肌酐、尿素氮,HE染色观察肾脏病理改变,TUNEL染色检测肾小管上皮细胞凋亡,western印迹检测肾组织Caspase-3表达。结果:建模后24h与A组相比,B组血清肌酐、尿素氮均明显升高(P<0.01);病理形态学检测提示模型大鼠发生明显肾间质水肿、肾小管上皮细胞胞浆空泡样变、肾小管上皮细胞凋亡(P<0.01),Caspase-3蛋白表达显著增多(P<0.01)。与B组相比,C、D组血清肌酐、尿素氮明显回落(P分别<0.01or<0.05),肾脏病理改变显著为轻,肾小管上皮细胞凋亡指数显著减少(P<0.01),Caspase-3蛋白表达明显降低(P<0.01)。结论:caspase3依赖的肾小管上皮细胞凋亡参与了大鼠造影剂肾病的发生,制大黄-川芎药对能通过抑制caspase3抑制肾小管上皮细胞凋亡,并进而保护CIN大鼠肾功能。

Objective： Explore effects and mechanism of Zhi Dahuang - Chuan Xiong Drug Pair on tubular epithelial cell ap- optosis in contrast - induced nephropathy （CIN） rats. Methods： Thirty - two male SD rats were divided into four groups （ n = 8）, nor- mal control group （group A）, CIN model group （group B）, Drug Pair with High Dose treatment group （group C）, and Drug Pair with Low Dose treatment group （group D）. Rats of group C and D were given daily intragastric administration with different doses of Drug Pair decoction separately at 7 days before building model. All rats were killed 24 h after injection of contrast media. The kidney injury was assessed by serum creatinine （Scr）, urea nitrogen （BUN）, and HE staining. Apoptosis of tubular epithelial cells was de- tected by TUNEL staining. Simultaneously, caspase3 expression in protein level was detected by western blotting method. Results： Twenty-four hours after injection of contrast media, compared with group A, Scr, BUN, and Caspase3 protein expression were sig- nificantly increased in group B rats （P 〈0.01 ）. Pathological changes of renal interstitium were the most serious in group B, which mainly showing vacuolar degeneration and apoptosis in tubular epithelial cells （P 〈0.01 ）. Compared with group B, Scr, BUN, and Caspase3 protein expression were significantly decreased in group C and D （P 〈 0.01 or 0.05 ）, which accompanied with lighter path- ological changes in renal interstitium and lower apoptosis index （AI） （ P 〈 0.01 ）. Conclusion ： Caspase3 depended apoptosis plays an important role in the pathogenesis of CIN rats. Zhi Dahuang - Chuan Xiong Drug Pair could inhibit such tubular epithelial cell apopto- sis by inhibiting Caspase3 expression, and thus beneficially protect rats again＇st contrast induced nephropathy.