基于对Klotho表达的调控作用探讨何首乌提取物二苯乙烯苷对慢性肾衰竭大鼠高脂血症的影响

Experimental Study on Effects of TSG Extracted from Heshouwu(Radix Polygoni Multiflori)on Hyperlipidemia Caused by Chronic Kidney Failure Rats by Regulating Klotho Expression

目的:探讨何首乌提取物二苯乙烯苷(TSG)对慢性肾衰竭大鼠肾脏Klotho蛋白表达及血脂变化的影响。方法:40只大鼠随机分为假手术组、模型组、TSG组[20 mg/(kg·d)]、缬沙坦组[55 mg/(kg·d)],每组10只。采用5/6肾脏切除法建立慢性肾衰竭模型,分别灌胃给药4周,假手术组与模型组给予等剂量饮用水灌胃。全自动生化分析仪检测大鼠血肌酐(SCr)、尿素氮(BUN)、总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C),及24 h尿蛋白定量(24 hUP),计算三酰甘油与高密度脂蛋白胆固醇比值(TG/HDL-C)。肾组织HE染色,光镜观察肾脏病理改变;免疫组化及Western blot检测肾组织Klotho蛋白及TGF-β1(转化生长因子-β1)表达。结果:与假手术组相比,模型组SCr、BUN及24 hUP水平均升高(均P<0.01);与模型组比较,TSG和缬沙坦组以上指标均下降(P<0.05或P<0.01),两给药组之间无明显差异(P>0.05)。模型组TC、TG、LDL-C、TG/HDL-C均明显高于假手术组(P<0.05或P<0.01),TSG组以上指标明显降低,与模型组相比有明显差异(P<0.05或P<0.01),缬沙坦无降低血脂作用。免疫组化和Western blot结果显示,模型组Klotho蛋白表达明显低于假手术组(P<0.01),TGF-β1表达明显高于假手术组(P<0.01);TSG及缬沙坦均可以升高Klotho蛋白表达(P<0.05或P<0.01),降低TGF-β1表达(P<0.05或P<0.01)。结论:TSG能调节慢性肾衰竭大鼠血脂异常,尤其是降低TG水平及TG/HDL-C,保护肾功能,其作用机制可能与升高Klotho蛋白表达相关。

Objective:To investigate the effects of Tetrahydroxy Stilbene Glucoside(TSG)extracted from Radix Polygoni Multiflori on Klotho protein expression and serum lipid in renal failure rats.Methods:40 Rats were randomly divided into sham-operation group,model group,TSG group and Valsartan group.5/6 nephrectomy was used to establish the model.TSG[20 mg/(kg·d)]and Valsartan[55 mg/(kg·d)]were administered intragastrically,the rats in sham operation group and model group were given the same dose of drinking water for 4 weeks.SCr,BUN,TC,TG,LDL-C,HDL-C and 24-hour urine protein(24 h UP)were detected by automatic biochemical analyzer,and calculate the ratio of TG/HDL-C.Renal pathological changes were observed by light microscopy,the protein expressions of Klotho and TGF-β1(transforming growth factorβ1)were detected by immunohistochemistry and Western blot.Results:Compared with sham-operation group,SCr,BUN and 24 hUP were increased(P<0.01)in model group and which were decreased in TSG and Valsarta groups(P<0.05 or P<0.01),but there was no significant difference between the TSG and Valsarta groups(P>0.05).The TC,TG,LDL-c,TG/HDL-C in the model group were significantly higher than those in the sham operation group(P<0.01 or P<0.05),which were significantly reduced in TSG group,(P<0.05 or P<0.01),however,blood lipid levels of Valsartan group were not reduced.The expression of Klotho protein was significantly decreased(P<0.01)in model group,while the expression of TGF-β1 was significantly increased(P<0.01).Klotho protein expression was increased in Both TSG and Valsartan groups(P<0.05 or P<0.01)and TGF-β1 expression was decreased(P<0.05 or P<0.01).Conclusion:TSG can modulate dyslipidemia,especially decrease TG and TG/HDL-C,and protect renal function in chronic kidney failure rats.The mechanism may be related to the increasing of Klotho protein expression.