女贞子-淫羊藿药对对顺铂所致大鼠肾毒性的保护作用

Profective effect of Ligustrum lucidum and Epimedium brevicornum on cisplatin induced nephrotoxicity in rat

目的探究女贞子-淫羊藿药对对顺铂所致大鼠肾毒性的保护作用。方法将Wistar雄性大鼠应用随机数字表法分为对照组、模型组、女贞子-淫羊藿药对组,每组10只。除对照组外,其他两组均于第4 d腹腔注射顺铂3 mg/kg;女贞子-淫羊藿药对组在给予顺铂前3 d灌胃350 mg/kg提取物,连续7 d。取材结束后,计算大鼠体重变化率和脏器指数,考察肾功指标和组织病理学变化,采用免疫组化法观察大鼠肾脏肾损伤分子1(Kidney injury molecule-1,KIM-1)和NADPH氧化酶4(Anti-NADPH oxidase 4,NOX4)阳性信号的表达和分布;Western Blot法检测KIM-1和NOX4蛋白表达的影响;试剂盒微量法检测大鼠肾组织中丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、还原型谷胱甘肽(reduced glutathione,GSH)和烟酰胺腺嘌呤二核苷磷酸/还原型辅酶II(NADP+/NADPH)水平的变化;ELISA法检测女贞子-淫羊藿药对对大鼠肾组织中白介素-1β(Interleukin-1β,IL-1β)、白介素-6(Interleukin-6,IL-6)和肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)水平的影响。结果相较于对照组,顺铂导致大鼠体重和肾组织SOD、CAT、GSH和TNF-α水平显著性降低(P<0.05),肾脏器指数、血清血尿素氮(blood urea nitrogen,BUN)、肌酐(creatinine,CREA)、尿酸(uric acid,UA)、肾组织MDA、NADP+/NADPH、IL-1β、IL-6水平显著性升高(P<0.05),HE染色显微镜下显示肾小管上皮细胞坏死脱落,管腔变形空泡化,KIM-1和NOX4阳性信号和蛋白表达均明显增加(P<0.05);相比于模型组,女贞子-淫羊藿药对干预使肾组织SOD、CAT、GSH和TNF-α水平显著升高(P<0.05),肾脏器指数、血清BUN、UA和肾组织MDA、NADP+/NADPH、IL-6水平显著下降(P<0.05),肾组织HE染色显示损伤减轻且KIM-1和NOX4阳性信号和蛋白表达均明显降低(P<0.05)。结论女贞子-淫羊藿药对通过抑制肾组织中KIM-1的表达,激活抗氧化酶的活性和抑制炎症因子的释放,发挥了对顺铂肾毒性的保护作用。

Objective To investigate the protective effect of combination of Ligustrum lucidum and Epimedium brevicornum(herb pair)on rat nephrotoxicity induced by cisplatin.Methods Wistar male rats were randomly divided into control group,model group and herb pair group,with 10 rats in each group.Except the control group,the other two groups were intraperitoneally injected with 3 mg/kg cisplatin on day 4;rats in herb pair group were given 350mg/kg extract intragastric administration 3 days before cisplatin for 7 consecutive days.After sampling,the change rate of body weight and organ index of rats were calculated,renal function indexes and histopathological changes were investigated.Immunohistochemistry was used to observe the expression and distribution of positive signals of kidney injury molecule–1(KIM-1)and anti-NADPH oxidase 4(NOX4)in the kidney of rats.Western Blot was used to detect the effects of KIM-1 and NOX4 protein expression.The changes of malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT),reduced glutathione(GSH)and NADP+/NADPH in rat kidney tissue were detected by kit microassay.The effects of herb pair on Interleukin-1b(IL-1b)、Interleukin-6(IL-6)and tumor necrosis factor-a(TNF-a)in kidney tissues of rats were determined by ELISA.Results Compared with the control group,cisplatin significantly reduced the body weight and levels of SOD,CAT,GSH,and TNF-αin the renal tissue of rats(P<0.05),The levels of renal organ index,serum blood urea nitrogen(BUN),creatinine(CREA),and renal tissue MDA,NADP+/NADPH,IL-1β,and IL-6 were significantly increased(P<0.05),HE staining microscope showed that the renal tubular epithelial necrosis were necrotic and exfoliated,and the lumen was deformed and vacuolized,KIM-1 and NOX4 positive signals and protein expression were significantly increased(P<0.05);Compared with the model group,the intervention of Ligustrum lucidum and Epimedium herb significantly increased the levels of SOD,CAT,GSH,and TNF-a in renal tissue(P<0.05).The renal organ index,serum BUN,UA,and renal tissue MDA,NADP+/NADPH,IL-6 levels significantly decreased(P<0.05).HE staining of renal tissue showed a reduction in damage and a significant decrease in KIM-1 and NOX4 positive signals and protein expression(P<0.05).Conclusion The combination of Ligustrum lucidum and Epimedium brevicornum exerted a protective effect against cisplatin-induced renal toxicity through inhibiting the expression of KIM-1 in kidney tissue,activating the activity of antioxidant enzymes,and suppressing the release of inflammatory factors.