氧化苦参碱对糖尿病大鼠肾组织纤维化的作用及可能机制研究

Effect of Oxymatrine on Renal Fibrosis in Diabetic Rats and Its Possible Mechanism

目的探讨氧化苦参碱(OM)对糖尿病(DM)大鼠肾组织纤维化的作用及可能机制。方法 SD大鼠随机分为正常对照(NC)组、糖尿病(DM)组、氧化苦参碱治疗(OM)组大鼠,尾静脉注射STZ复制DM模型,于成模13周起,DMA组大鼠采用OM个体化治疗,以血糖控制在4~7mmol/L,尿糖阴性为准,分别于24周处死各组大鼠。生化方法测血糖、血肌酐和24h尿蛋白;HE和Masson染色观察大鼠肾组织结构改变;免疫组化检测各组大鼠肾组织转化生长因子-β1(TGF-β1)、E-钙黏附素(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)和纤维连接蛋白(FN)蛋白的表达变化;免疫荧光检测大鼠肾组织中Arkadia和核转录共抑制因子(SnoN)蛋白的表达。结果与NC组相比,DM组大鼠血糖,血肌酐,24h尿蛋白明显增多(P<0.05),并出现肾组织形态学异常改变;而OM组大鼠血糖、血肌酐、24h尿蛋白均显著低于DM组(P<0.05),肾纤维化病变明显改善;DM组TGF-β1、α-SMA和FN蛋白表达显著增加(P<0.05)、E-cadherin蛋白表达显著减少(P<0.05);与DM组相比,OM组TGF-β1、α-SMA和FN蛋白表达显著减少(P<0.05),E-cadherin蛋白表达显著增加(P<0.05);免疫荧光显示Arkadia与SnoN蛋白的表达趋势相反。结论 OM具有抑制DM大鼠肾组织纤维化的作用,其机制可能与通过抑制TGF-β1表达和抑制Arkadia泛素化降解SnoN蛋白有关。

Objective To investigate the effects of oxymatrine（OM） to improve diabetes（DM） renal fibrosis in rats,and study its possible mechanism. Methods SD rats were randomly divided into control（NC）, diabetes mellitus （DM） and OM-treated DM groups（OM）. DM model of rat was made by injecting streptozotocin（STZ） via tail vein. The blood glucose was controlled to the level of 4~7 retool/L, and urine glucose remained negative. The rats were sacrificed at 24 h week respectively. The blood glucose, serum creatinine and 24 h urine protein were measured by biochemical method. The pathological changes of renal tissue by HE and Massion staining were observed under light microscope. Immunohistochemistry was employed to assay the expression of TGF-β1, FN, E-eadherin and α-SMA protein in the rat renal tissue. In addition,the expression of Arkadia and Ski-related novel protein N（SnoN） was also examined by Immunofluorescence. Results Compared with NC group, the blood glucose, serum creatinine, and 24 h u- rine protein of DM group increased significantly（P〈0. 05）, and renal tissue were presenting typical morphologic changes at different time points. The blood glucose,serum ereatinine,and 24 h urine protein of OM group were significantly lower than those in DM group（P〈0.05）. Compared with NC group,the rats of group DM appeared in renal tissue fibrosis,TGF-β1,α-SMA and FN protein expression increased significantly（P〈0, 05）, E-cadherin protein expression were significantly decreased（P〈0.05）. Compared with DM group,OM in renal tissue of rats fibrosis significantly improved,TGF-β1, α-SMA and FN protein expression were significantly decreased（P〈0.05）, the E-cadherin protein expression increased significantly（P〈0.05）. Immunofluoreseence showed the expression tendency of Arkadia and SnoN protein in contrast. Conclusion OM can ameliorat renal fibrosis in DM rats through relieving ex- pression of TGF-β1 protein associated with inhibition of Arkadia ubiquitination and degradation of SnoN protein.