冬虫夏草对糖尿病大鼠肾小管上皮细胞ILK表达的影响

Effects of Cordyceps Sinensis on Expression of ILK in Kidney of Diabetic Rats

【目的】观察糖尿病大鼠肾小管上皮细胞整合素连接激酶(ILK)的表达及冬虫夏草对其表达的影响。【方法】雄性Wistar大鼠随机分为3组:正常对照组(N组,n=12)、糖尿病模型组(D组,n=12)、糖尿病冬虫夏草干预组[C组,n=12,冬虫夏草1000mg/(kg·d)灌胃]。成模后第8周末及16周末各组分别处死6只大鼠,测定24h尿蛋白排泄量、血肌酐;HE及MASSON染色法观察肾脏病理改变;免疫组织化学法检测ILK、E钙粘蛋白(E-cad)、α平滑肌肌动蛋白(α-SMA)及转化生长因子β1(TGF-β1)的表达。【结果】①与N组比较,D组大鼠肾小管间质损伤指数、肾间质胶原面积明显增加(P<0.01)。②D组大鼠肾小管上皮细胞E-cad阳性表达显著低于N组,TGF-β1、ILK和α-SMA阳性表达均显著高于N组;E-cad表达和TGF-β1表达呈负相关(rs=-0.60,P<0.05);ILK、α-SMA表达和TGF-β1表达呈正相关(rs=0.88,P<0.01;rs=0.91,P<0.05)。③C组大鼠肾小管间质损伤指数、肾间质胶原面积较D组明显减弱,其肾小管上皮细胞的E-cad的表达较D组明显增加,TGF-β1、ILK和α-SMA表达较D组均明显减弱(P<0.01)。【结论】冬虫夏草能通过下调ILK表达发挥肾脏保护作用。

[Objective]To study the effect and mechanism of Cordyceps sinensis on the process of tubular epithelial-mesenehymal transformation in kidney of diabetic rats. [Methods] Wistar male rats were randomly assigned to 3 groups： control group（N group, n = 12）, diabetic group（D group, n = 12）, Cordyceps sinensis treatment group（C group, n =12）. The rats in Cordyceps sinensis treatment group were treated with daily intragastric administration of 1000mg/（kg · d） of Cordyceps sinensis after the models were successfully established. Six rats of each group were killed respectively at 8 and 16 weeks after experiment. The excretion of urinary protein and serum creatine were measured. Renal histological changes were observed by HE and MASSON staining. Immunohistochemistry was performed to investigate the expressions of TGF-β1, ILK, E-cadherin and α-SMA in kidney, [Results]①Compared with the control group, the tubulointerstitial injury and the accumulation of extracellular matrix protein in diabetic models were obvious（ P〈0.01）; ② Compared with the control group, the expression of E-cad was decreased significantly and the expressions of TGF-β1, ILK and α-SMA were increased significantly in diabetic group. E-cad was negatively correlate to TGF-β1 （rs = -0.60, P 〈0.05）, and ILK and α-SMA were positively correlate to TGF-β1 （rs =0.88, P 〈0.01;rs =0.91. P 〈0. 05）; ③In comparison with diabetic group, Cordyceps sinensis could up-regulate the expression of E-cad and down-regulate the expressions of TGF-β1 , ILK and α-SMA in renal tubular epithelial cells （ P 〈0.01）. [Conclusion] Cordyceps sinensis could inhibit tubular epithelial-mesenchymal transformation（EMT）, protect kidney by ameliorating interstitial fibrosis in diabetic rats. The mechanism may depend on downregulating the expressions of TGF-β1 and ILK.