芦荟大黄素对小鼠肾毒性的作用机制

Mechanism of Nephrotoxicity of Aloe Emodin in Mice

目的:研究长期给予不同剂量的芦荟大黄素导致小鼠肾毒性,并探讨其毒性机制。方法:将30只昆明种小鼠随机分为雌雄空白组、雌雄芦荟大黄素低、高剂量组(0.8,1.6 g·kg^(-1))。芦荟大黄素各剂量组连续灌胃给药11周,每日早晚各1次。采用生化试剂盒检测小鼠血清尿素氮(urea nitrogen,BUN),肌酐(creatinine,SCr),超氧化物歧化酶(superoxide dismustase,SOD)和丙二醛(malondialdehyde,MDA)含量,肾脏总谷胱甘肽(total glutathione,T-GSH)/氧化性谷胱甘肽试剂盒(oxidized glutathione,GSSG)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平;酶联免疫吸附试验(ELISA)检测血清中肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)和白细胞介素-6(interleukins-6,IL-6)水平;苏木素-伊红(HE)染色检测肾脏病理变化;免疫组化检测肾脏半胱氨酸蛋白酶-3(cysteine aspartic acid specific protease-3,Caspase-3)和转化生长因子-β_1(transforming growth factor-β_1,TGF-β_1)蛋白表达。结果:与同性别空白组比较,芦荟大黄素高剂量组雌、雄小鼠血清中BUN含量升高(P<0.05,P<0.01),芦荟大黄素高剂量组雄性小鼠血清中SCr含量升高(P<0.05),芦荟大黄素低剂量组肾小管轻度损伤,高剂量肾小管和肾小球中度损伤;与空白组比较,芦荟大黄素高剂量组雌、雄小鼠血清MDA含量升高(P<0.05),SOD活性降低(P<0.05),芦荟大黄素高剂量组雌雄小鼠肾脏中GSH/GSSG含量降低(P<0.05),雄性小鼠Caspase-3蛋白表达增加(P<0.05);与空白组比较,芦荟大黄素高剂量组雄性小鼠TNF-α和IL-6含量升高(P<0.05),芦荟大黄素低、高剂量组雌雄小鼠肾脏TGF-β_1蛋白表达均增加(P<0.05)。结论:芦荟大黄素1.6 g·kg^(-1)给药11周,对小鼠肾脏有毒性作用,其机制与机体氧化应激、细胞凋亡和TGF-β_1蛋白表达有关。

Objective:To study nephrotoxicity induced by long-term administration of different doses of aloe-emodin in mice,and explore its mechanism.Method:A total of 30 male and female Kunming mice were randomly divided into normal control group,and low-dose aloe-emodin group,high-dose aloe-emodin group(0.8,1.6 g·kg-1).Every dose of group was administered intragastrically for 11 weeks,twice daily.effect of serum urea nitrogen(BUN),creatinine(SCr),superoxide dismutase(SOD),malondialdehyde(MDA),Glutathione(GSH/GSSG)and Glutathione Peroxidase(GSH-Px)levels were detected by biochemical kits according to manufacturer’s instruction.Enzyme-linked immune assay was used to determine serum tumor necrosis factor(TNF)-αand interleukins(IL)-6 levels.Hematoxylin eosin(HE)staining was used to detect renal pathological changes in kidney tissues,and cysteine aspartic acid specific protease(Caspase)-3 and transforming growth factor(TGF)-β1proteins were determined by immunohistochemistry.Result:According to results,compared with normal control group,the levels of BUN and SCr in serum with high-dose aloe-emodin were increased.The renal tubules in low-dose group were mildly injured,while renal tubules and glomeruli of high-dose group were moderately damaged.Compared with normal control group,the level of SOD was significant decreased(P<0.05),MDA was increased(P<0.05),the levels of GSH/GSSG in kidneys of high-dose groups were decreased(P<0.05).In high-dose group,the expression of Caspase-3 protein was increased in kidneys,especially in males(P<0.05).Compared with normal control group,the levels of TNF-αand IL-6 were increased,the expression of TGF-β1protein in kidneys was increased in low-dose and high-dose groups(P<0.05).Conclusion:results show that 1.6 g·kg-1aloe-emodin was administered intragastrically for 11 weeks,which had toxic effects on kidney in mice.The mechanism may be related to oxidative stress,apoptosis and TGF-β1protein expression.