补气健脾益肾法对糖尿病肾病大鼠肾小管上皮细胞转分化及肾间质纤维化影响

Effect of Buqi Jianpi Yishen Method on Renal Tubular Epithelial Cell Transdifferentiation and Renal Interstitial Fibrosis for Diabetic Nephropathy Rats

目的:分析补气健脾益肾法对糖尿病肾病(DN)大鼠肾小管上皮细胞的间充质转分化(EMT)及肾间质纤维化影响。方法:选取SPF级Wistar雄性大鼠45只,随机数字表法将分成三组,正常组、模型组和观察组,每组各15只,观察组和模型组制备DN模型,观察组大鼠使用补气健脾益肾法灌胃,剂量为1.4g/kg,正常组和模型组大鼠使用同剂量生理盐水灌胃,每天1次,共灌胃8周。末次给药后大鼠置入代谢笼内,采集其24小时排泄尿液,计算其24小时尿量,全自动生化分析仪检测24小时尿蛋白、血尿素氮及血肌酸酐含量,血糖仪检测其血糖含量,肾组织常规行HE染色,在光镜下查看大鼠肾脏组织的病理状况,免疫组化检测肾组织内转化生长因子-β(TGF-β)、骨成形蛋白(BMP-7)及α-平滑肌肌蛋白(α-SMA)表达状况。结果:模型组大鼠24h尿蛋白、肾质量及24h尿量比正常组上升,观察组24h尿蛋白、肾质量及24h尿量比模型组下降,差异均有统计学意义(P<0.05);模型组及观察组血糖、模型组血尿素氮、血肌酐酸含量比正常组上升,观察组血尿素氮、血肌酐酸比模型组下降,差异均有统计学意义(P<0.05);模型组大鼠BMP-7表达量比正常组下降,TGF-β、α-SMA表达量比正常组上升,观察组BMP-7表达量比模型组上升,TGF-β、α-SMA表达量比模型组下降,差异均有统计学意义(P<0.05)。结论:补气健脾益肾法可降低DN大鼠尿蛋白含量,抑制肾小球EMT和肾间质纤维化发生发展。

Objective:To analyze the effect of Buqi Jianpi Yishen Recipe on mesenchymal transdifferentiation(EMT)and renal interstitial fibrosis in renal tubular epithelial cells of diabetic nephropathy(DN)rats.Methods:A total of 45 male Wistar male rats of SPF grade were randomly divided into three groups:normal group,model group and observation group,with 15 rats in each group.The observation group and the model group were prepared with DN model.Buqi Jianpi Yishen Recipe was intragastrically administered at a dose of 1.4 g/kg.The rats in the normal group and the model group were intragastrically administered with the same dose of normal saline once a day for a total of eight weeks.After the last administration,the rats were placed in a metabolic cage,and the urine was collected for 24 hours to calculate the urine volume.The 24 h urine protein,blood urea nitrogen and blood creatinine content were detected by automatic biochemical analyzer,and the blood glucose level was detected by blood glucose meter.Renal tissue was routinely stained with HE.The pathological condition of rat kidney tissue was observed under light microscope.Immunohistochemistry was used to detect the expression status of transforming growth factor-β(TGF-β),bone forming protein(BMP-7)andα-smooth muscle myoprotein(α-SMA).Results:The 24 h urine protein,kidney mass and 24 h urine volume of the model group were higher than those of the normal group.The 24 h urine protein,kidney mass and 24 h urine volume of the observation group were significantly lower than the model group(P<0.05).The blood glucose and blood creatinine levels in the model group and the observation group were higher than those in the normal group.The blood urea nitrogen and serum creatinine ratio in the observation group decreased,and the difference was statistically significant(P<0.05).The expression of BMP-7 in rats was lower than that in the normal group.The expression of TGF-βandα-SMA was higher than that in the normal group.The expression of BMP-7 in the observation group was higher than that in the model group,and the expression of TGF-βandα-SMA was higher than that in the model group.The difference was statistically significant(P<0.05).Conclusion:Buqi Jianpi Yishen method can reduce the urine protein content of DN rats and inhibit the development of glomerular EMT and renal interstitial fibrosis.