益气养阴清热方对糖尿病肾病小鼠保护作用及机制探索

Effect and Mechanism of Yiqi Yangyin Qinre Decoction(益气养阴清热方)on Diabetic Nephropathy Mice

目的 通过观察益气养阴清热方对糖尿病肾病小鼠肾组织保护作用,进而探讨其可能的作用机制。方法20只db/db小鼠随机分为模型组、益气养阴清热方治疗组,10只db/m小鼠作为空白对照组。给药12周后,采集小鼠血尿及组织标本,检测血肌酐(Scr)、血尿素氮(BUN)、血糖(Glu)、尿白蛋白/肌酐(ACR);HE染色、PAS染色并通过光镜观察小鼠肾组织病理改变;通过电镜观察小鼠肾组织超微结构改变情况;Western blot检测小鼠肾组织IL-1β、IL-6、IL-18、NF-κB、Beclin-1、Atg-7、LC3、p62的蛋白表达水平。结果 与对照组比较,模型组小鼠出现肾小球体积增大、炎性细胞浸润、肾小球基底膜增厚,系膜区扩张,系膜基质增多、足突融合变形等宏观及微观病理改变,血清Scr、BUN、Glu和ACR增加(P<0.01),肾脏IL-1β、IL-6、IL-18、NF-κB、p62蛋白表达水平上升(P<0.01),Beclin-1、Atg-7、LC3蛋白表达水平下降(P<0.01);与模型组相比,治疗组经中药干预后,肾脏病理损伤均有改善,Scr、BUN、Glu和ACR显著降低(P<0.01),同时组织中IL-1β、IL-6、IL-18、NF-κB、p62蛋白表达减少(P<0.01),Beclin-1、Atg-7、LC3蛋白表达增加(P<0.01)。结论 益气养阴清热方具有调节血糖、保护肾功能的作用,其机制可能与调控自噬通路、抑制炎症反应相关。

Objective To observe the protective effect of Yiqi Yangyin Qingre Decoction(益气养阴清热方)on renal tissue of diabetic nephropathy mice and explore its possible mechanism.Methods 20 db/db mice were randomly divided into model group,Yiqi Yangyin Qingre Decoction treatment group and 10 db/m mice as blank control group.After 12 weeks of administration,hematuria and tissue samples were collected to detect serum creatinine(Scr),blood urea nitrogen(BUN),blood glucose(Glu),urinary albumin/creatinine(ACR);HE staining,PAS staining and through light microscope to observe the pathological changes of kidney tissue;ultrastructural changes of kidney tissue were observed by electron microscope.The protein expressions of IL-1β,IL-6,IL-18,NF-κB,Beclin-1,Atg-7,LC3 and p62 in kidney tissue of mice were detected by Western blot.Results Compared with the control group,the model group showed macroscopic and microscopic pathological changes,such as glomerular volume increase,inflammatory cell infiltration,glomerular basement membrane thickening,mesangial expansion,mesangial matrix increase,foot process fusion and deformation,serum Scr,BUN,Glu and ACR increased(P<0.01),and renal IL-1β,IL-6,IL-18,NF-κB,p62 protein expression increased(P<0.01).The protein expression of Beclin-1,Atg-7 and LC3 decreased(P<0.01).Compared with the model group,the renal pathological injury in the treatment group was improved,Scr,BUN,Glu and ACR were significantly decreased(P<0.01),and the expression of IL-1β,IL-6,IL-18,NF-κB,p62 protein decreased(P<0.01)and the expression of Beclin-1,Atg-7,LC3 protein increased in the treatment group(P<0.01).Conclusion Yiqi Yangyin Qingre Decoction can regulate blood glucose and protect renal function,and its mechanism may be related to the regulation of autophagy pathway and inhibition of inflammation.