灵芝多糖对糖尿病肾病小鼠肾小管上皮细胞间充质转化的抑制作用

Inhibitory effect of ganoderma lucidum polysaccharide on renal tubular endothelial-to-mesenchymal transition of diabetic nephropathy mice

目的探讨灵芝多糖对糖尿病肾病(DN)小鼠肾小管上皮细胞间充质转化(EMT)的影响。方法将40只C57BL/KsJ-db/db小鼠按体重随机分为5组,每组8只。正常组给予普通饲料喂养。模型组、灵芝多糖(GLP)低(100 mg/kg)、高(400 mg/kg)剂量组和厄贝沙坦组(50 mg/kg)给予高糖饲料喂养,DN小鼠造模成功后,各组每日灌胃给药,持续8周,测定各组小鼠肾功能、血脂等生理指标变化;取肾组织HE染色观察GLP对小鼠肾脏病理改变的影响,用Western blot和实时荧光定量PCR实验检测E-cadherin、α-SMA、TGF-β1、Fibronectin、Vimentin、Ncadherin、p-Smad2、p-Smad3、Smad2、Smad3蛋白表达及mRNA水平。结果与模型组比较,GLP低、高剂量组血脂代谢及肾功能升高(P <0.05)。与模型组比较,GLP低、高剂量组及厄贝沙坦组E-cadherin蛋白及mRNA水平升高,而α-SMA、Fibronectin、Vimentin、N-cadherin蛋白及mRNA水平降低(P<0.01)。Smad2、Smad3总蛋白表达水平无变化。结论GLP可能通过调节TGF-β1/Smad信号通路,抑制DN小鼠的EMT。

Objective To investigate the effect of ganoderma lucidum polysaccha-ride(GLP) on epithelial-to-mysenchymal transition(EMT) of renal tubular epithelial cells in diabetic nephropathy(DN) mice.Methods Forty C57 BL/KsJ-db/db mice were randomly divided into 5 groups by weight.Normal group was fed with normal feed.Model group,GLP low(100 mg/kg),high(400 mg/kg) dose groups and Irbesartan group(50 mg/kg,Irbesartan) were given high-sugar dieat.Each group was given the drug by gavage daily for 8 weeks after successful modeling of DN mice.The changes of renal function,blood lipid and other physiological indexes were measured.The protein expression and mRNA level of E-cadherin,α-SMA,TGF-β1,Fibronectin,Vimentin,N-cadherin,TGF-β1,p-Smad2,p-Smad3,Smad2 and Smad3 were measured by Western Blot and Real-time PCR.Results Compared with the model group,the renal function and blood lipid metabolism in GLP low and high doses group were significantly decreased(P <0.05).Compared with the model group,the levels of E-cadherin protein and mRNA were increased in the GLP low and high doses group and the Irbesartan group,while the levels of α-SMA,Fibronectin,Vimentin,N-cadherin protein and mRNA were decreased(P <0.01).The expression levels of Smad2 and Smad3 were unchanged.Conclusion GLP inhibits EMT maybe via inhibition of TGF-β1/Smad signaling pathway.