肾炎康血清干预嘌呤霉素氨基核苷诱导足细胞损伤研究

摘要

Objective:To investigate the protective effect of Compound ShenYanKang Granules,a Chinese herbal recipe mainly composed of Galium aparine,Panax notoginseng,Coicis Semen,Herba Pyrolae,on the podocyte injury induced by Puromycin amino nucleside. Method:The primary-cultured podocytes of rats were randomly divided into normal group,control group,low-dose,middle-dose, high-dose groups of ShenYanKang and irbesartan group. The podocytes in the normal group were given normal rat serum alone while that of the other groups were all given serum containing Puromycin amino nucleoside, and additionally,ShenYanKang groups and irbesartan group were given serum containing low-dose,middle-dose and high-dose ShenYanKang and irbesartan respectively for 48 hours. The apoptosis of podocyte was determined by TUNEL assay. P53 and Caspase-3 mRNA were observed by real-time PCR. Expression of p53 and Caspase-3 protein were examined by Western blot. Result:The apoptosis rate of podocytes,P53 and Caspase 3 mRNA expression level in Puromycin amino nucleoside control group were significantly higher than that in the normal group(P<0.05 or P<0.01),and were decreased in the medication groups(P<0.05 or P<0.01 compared with that in Puromycin amino nucleoside control group).The apoptosis rate of podocytes,P53 and Caspase 3 expression level in high-dose ShenYanKang group was not significantly different from that in irbesartan group(P>0.05).Conclusion: Compound ShenYanKang could inhibit the podocyte apoptosis induced by Puromycin amino nucleoside and its mechanism may be related to the inhibition of P53 and Caspase 3 expression.%目的：探讨复方肾炎康对嘌呤霉素氨基核苷诱导足细胞损伤的保护作用。方法：原代培养大鼠足细胞，随机分为正常组，对照组，肾炎康低、中、高剂量组，厄贝沙坦组除正常组外，其余各组分别予以嘌呤霉素氨基核苷，正常组加正常大鼠血清，肾炎康低、中、高剂量组分别加肾炎康低、中、高剂量，厄贝沙坦组加厄贝沙坦，均干预48h。用原位末端标记法（TUNEL）检测细胞凋亡，实时定量PCR方法检测P53、Caspase 3mRNA，western印迹检测细胞中P53、Caspase 3蛋白变化。结果：嘌呤霉素氨基核苷作用下的各组足细胞凋亡率、P53、Caspase 3mRNA及蛋白表达均显著高于正常组（P＜0.05，P＜0.01），药物干预的各组足细胞凋亡率、P53、Caspase 3mRNA及蛋白表达均显著低于对照组（P＜0.05，P＜0.01），肾炎康高剂量组足细胞凋亡率、P53、Caspase 3mRNA及蛋白表达与厄贝沙坦组比较差异无统计学意义（P＞0.05）。结论：中药复方肾炎康能抑制嘌呤霉素氨基核苷诱导的足细胞凋亡，其机制可能与抑制P53、Caspase 3表达有关。