心肝宝胶囊对STZ诱导的糖尿病大鼠早期肾损伤的保护作用及其机制研究

作者	单位
高洁	山东大学齐鲁医院肾内科
范例	山东大学齐鲁医院肾内科
刘茂静	山东大学齐鲁医院肾内科
甄军晖	滨州医学院附属医院肾内科
姜虹	山东大学齐鲁医院教育部和卫生部心血管重构与功能研究重点实验室
胡昭	山东大学齐鲁医院肾内科

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中文摘要:

目的探讨心肝宝胶囊对链尿佐菌素(streptozocin,STZ)诱导的糖尿病(diabetes mellitus,DM)大鼠早期肾损伤的保护作用及其机制。方法选取24只雄性Wistar大鼠建立STZ DM动物模型。造模后随机分为模型组、心肝宝胶囊治疗组[中药组,0.5g/(kg·d)]及贝那普利治疗组[西药组,4mg/(kg·d)],每组8只,另选取8只大鼠作为空白对照组(空白组)。模型组与空白组给予等量自来水灌胃,连续干预8周。干预4、8周末检测各组大鼠血糖,8周后观察24h尿蛋白(24h urine protein,24hUP)、血尿素氮(blood urea nitrogen,BUN)和肌酐(serum creatinine,SCr),采用免疫组化法检测转化生长因子-β1(transforming growth factor-beta1,TGF-β1)、层黏连蛋白(laminin,LN)、Ⅳ型胶原蛋白(collagenⅣ,Col-Ⅳ)的表达,采用RT-PCR法检测肾脏TGF-β1、组织金属蛋白酶抑制剂-1(tissue inhibitor of matrix metalloproteinase-1,TIMP-1)及纤溶酶原激活物抑制剂(plasminogen activator inhibitor-1,PAI-1)mRNA的表达,行HE、PAS和Masson染色观察肾脏病理变化。结果与空白组比较,DM大鼠造模后出现血糖升高、多饮、多食、多尿、体重减轻、消瘦、体毛枯燥无光泽、易激惹等表现,干预8周后两个用药组症状减轻。与空白组比较,模型组24hUP、SCr、血糖、Col-Ⅳ、LN、TGF-β1阳性表达量,TGF-β1、TIMP-1及PAI-1mRNA,肾小球面积(GA)、细胞外基质(ECM)及ECM/GA均升高,差异均有统计学意义(P<0.01),病理改变可见明显肾小球体积增大,毛细血管袢扩张,小球系膜细胞增生,系膜区基质增多,增宽,基底膜增厚,肾小管排列紊乱,部分上皮细胞可见空泡变性、脱落,可见蛋白管型,间质有炎性细胞浸润。与模型组比较,两个用药组各指标均降低,差异均有统计学意义(P<0.01),病理改变较轻,系膜细胞轻度增生,系膜基质轻度增多。中药组SCr、PAI-1mRNA表达低于西药组(P<0.05),两个用药组其他指标比较,差异无统计意义(P>0.05)。结论心肝宝胶囊对STZ诱导的糖尿病大鼠早期肾损伤有一定保护作用及抗肾小球纤维化作用,其机制可能与下调TGF-β1、TIMP-1、PAI-1及Col-Ⅳ的表达,减少细胞外基质,减少尿蛋白有关。

英文摘要:

Objective To discuss the protective effects of Xin’ganbao Capsule(XC) on early kidney injury in streptozocin(STZ)-induced diabetic rats and its mechanisms.Methods Twenty-four male Wistar rats were selected to establish STZ induced diabetes mellitus(DM) model.After modeling they were randomly divided into the model group,the XC group(at the daily dose of 0.5 g/kg),and the benazepril group(at the daily dose of 4 mg/kg),8 in each group.Another 8 rats were chosen as the blank control group.Rats in the model group and the blank control group were administered with equal volume of normal saline by gastrogavage for 8 successive weeks.The blood glucose was monitored by the end of the 4th week and the 8th week.The 24 h urine protein(24 hUP),blood urea nitrogen(BUN),and serum creatinine(SCr) were detected by the end of the 8th week.The transforming growth factor-beta1(TGF-β1),laminin(LN),collagen Ⅳ(Col-Ⅳ) expression were detected using immunohistochemical assay.The mRNA expressions of renal TGF-β1,tissue inhibitor of matrix metalloproteinase-1(TIMP-1),and plasminogen activator inhibitor-1(PAI-1) were detected using RT-PCR.The pathological changes of the renal tissue were observed by HE,PAS,and Masson stain methods.Results Compared with the blank control group,hyperglycemia,polydipsia,polyphagia,polyuria,weight loss,emaciation,dry and dim body hair,and irritability appeared in the diabetic rats.After 8 weeks the symptoms of the two medication groups were attenuated.When compared with the blank control group,the 24 hUP,SCr,blood glucose,Col-Ⅳ,LN,TGF-β1 positive expression ratio,the levels of TGF-β1,TIMP-1,PAI-1 mRNA,the area of glomerular(GA),extracellular matrix(ECM),and ECM/GA all increased in the model group with statistical difference(P<0.01).The pathological changes showed obvious glomerular enlargement,the capillary loop expansion,the proliferation of the mesangial cells,increased mesangial matrix,widen and thicken glomerular basement membrane(GBM),and tubular derangement.The vacuolar degeneration and shedding could be seen in partial epithelial cells.The protein cast could also be seen with infiltration of interstitial inflammatory cells.Compared with the model group,each index of the two medication groups decreased with statistical difference(P<0.01).The pathological changes were less in the two medication groups.The mesangial cells were slightly proliferated and the mesangial matrix slightly increased.The mRNA expressions of SCr and PAI-1 were lower in the XC group than in the benazepril group(P<0.05).There was no statistical difference in the other indices between the two medication groups(P>0.05).Conclusions XC had some protective effects and anti-glomerulosclerosis effects on early kidney injury in STZ-induced diabetic rats.Its mechanisms might be associated with down-regulating the mRNA expressions of TGF-β1,TIMP-1,PAI-1,and Col-Ⅳ,reducing ECM and urine protein.

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