葛根素注射液对糖尿病KKA^y小鼠肾小管上皮细胞的影响

Effect of puerarin injection on epithelial-mesenchymal transition in renal tubular epithelial cells of diabetic KKA^y mice

目的:观察葛根素治疗糖尿病小鼠肾间质纤维化(renal interstitial fibrosis,RIF)过程中肾小管上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白表达的变化,为临床药物治疗糖尿病RIF提供实验依据。方法:将16只2型糖尿病模型KKAy小鼠随机分为模型组(n=8)和葛根素注射液治疗组(n=8)。8只C57BL/6J小鼠作为正常组。用药组小鼠14周龄开始以腹腔注射的方式给药,观测小鼠日常状态及血糖变化,于24周龄处死小鼠,取肾脏分别观察肾组织病理形态学变化,免疫组织化学法检测小鼠肾组织中α-平滑肌肌动蛋白(alpha-smooth muscle actin,α-SMA)、转化生长因子β1(transforming growth factorβ1,TGF-β1)和转化生长因子βI型受体(TGF-β-RI)蛋白的表达。结果:形态学观察可见模型组发生明显的纤维化改变,而经葛根素注射液治疗后,小鼠肾间质基质轻度增多,病变较轻微。治疗组KKAy小鼠肾组织中较模型组小鼠肾组织中α-SMA、TGF-β1和TGF-β-RI蛋白的表达减少。结论:葛根素注射液可在一定程度上减少α-SMA的表达,抑制KKAy小鼠肾脏组织中TGF-β1和TGF-β-RI蛋白表达,阻断并逆转EMT的过程,延缓肾间质纤维化的发生和发展。

AIM： To observe the effect of puerarin injection on the expression of epithelial-mesenchymal tran- sition-related proteins in KKAy mice with renal interstitial fibrosis （RIF）. METHODS： Sixteen KKAy mice were randomly divided into model group （n = 8） and puerarin injection treatment group （n = 8 ）, and 8 C57BL/6J mice were used as nor- real controls. The mice in treatment group were intraperitoneally given puerarin injection from the 14th week. The blood glucose levels were observed on a daily basis. The mice were sacrificed at the 24th week. The renal pathological changes were observed under light microscope. The expression of a-smooth muscle actin （ot-SMA）, transforming growth factor ~t （TGF-~,） and transforming growth factor ~ type I receptor （TGF-^-RI） in the renal tissues were examined by immunohis- tochemical staining. RESULTS ： Fibrosis was found in the KKAy mice of model group, while the mice in treatment group showed a slight increase in renal interstitium. Treatment with puerarin injection decreased the protein expression levels of α-SMA, TGF-β1 and TGF-β-RI in the kidney tissues as compared with those in model group. CONCLUSION： Puerarin injection reduces the expression of α-SMA, and restrains the protein expression of TGF-β1 and TGF-β-RI in the kidney tis- sue of KKAy mice. These changes may inhibit and reverse the process of epithelial-mesenchymal transition, thus delaying the occurrence and development of RIF.