“益气养阴通络法”通过调控SIRT1延缓糖尿病肾病进展的研究

Therapy with "Replenishing Qi,Nourishing Yin and Dredging Collaterals" Prevents the Progression of Diabetic Nephropathy by Up - regulating SIRT1

目的:观察SIRT1在"益气养阴通络法"治疗糖尿病肾病(diabetic nephropathy,DN)中的作用。方法:将179例确诊为Ⅲ期和Ⅳ期DN患者随机分为基础治疗对照组(76例)和"益气养阴通络法"综合治疗组(103例),评价两组患者临床症状疗效、尿微量白蛋白(Alb)及肌酐清除率(Ccr);并通过免疫组化和免疫荧光法分析SIRT1在人肾组织中的表达情况及在DN患者中的变化情况;进一步通过逆转录病毒沉默原代大鼠肾小管上皮细胞(TEC)SIRT1后,观察TEC在AGE-BAS刺激后的凋亡情况。结果:经过6个月治疗,"益气养阴通络法"综合治疗组中医症状积分改善、微量白蛋白排泄率降低及肌酐清除率升高的患者比率明显高于基础治疗对照组(P<0.01);停药6个月后进行随访,"益气养阴通络法"综合治疗组的远期疗效也优于基础治疗对照组(P<0.01)。免疫组化和免疫荧光显示,SIRT1在人肾组织的足细胞、内皮细胞及小管上皮细胞等均有表达,并且上述表达在DN患者中明显降低;而经"益气养阴通络法"治疗后SIRT1表达明显增强。进一步通过体外实验证实,沉默SIRT1后,cleaved caspase3蛋白表达在AGE-BSA刺激的TEC中明显升高。结论:"益气养阴通络法"在治疗DN过程中有着显著作用,而上述疗效可能是通过上调肾脏固有细胞中SIRT1实现的。

Objective：To observe the effect of SIRT1 during the treatment of on the therapy for diabetic nephmpathy （DN） through ＂Replenishing qi, nourishing yin and dredging collaterals＂. Methods：179 Ⅲ and Ⅳ DN patients were randomly divided into control group with basic treatment （76 cases） and investigation group with the combination of basic treatment and ＂replenishing qi, nourishing yin and dredging collaterals＂ therapy （103 cases）. Clinical symptoms, albumin excretion and creatinine clearance rate were evaluated. The expression of SIRT1 in human renal tissue and the change of SIRTI expression in DN patients were studied by immunohistochemical and immunofluorescent staining. The apoptosis of rat primary tubular epithelial cells （TECs） was assessed after SIRTI knockdown by lentivirus targeting SIRT1. Results： After 6 - month treatment, the remission rates in the clinical symptom score, albumin excretion and creatinine clearance were much higher in combined treatment group than basic treatment control group （P 〈0. O1 ） When follow - up at the 6th month after withdrawing the therapy, it was show that the long - term efficacy in the combined treatment group was also superior to the basic treatment control group （P 〈 0.01 ）. Furthermore, SIRT1 was found expressed in the podocytes, endothelial cells of glomeruli and epithelial cells i of tubules in human renal tissues by immunohistochemistry and immunofluorescence staining. Interestingly, the expression of SIRT1 was significantly decreased in DN patients compared with minimal change nephrotic syndrome patients. And the SIRT1 expression was up - regulated by ＂replenishing qi, nourishing yin and dredging collaterals＂ therapy. We also found that silencing of SIRT1 significantly enhanced the expression of cleaved caspase 3 in AGE - BSA treated TECs in vitro. Conclusion：The ＂replenishing qi, nourishing yin and dredging collaterals＂ therapy played a key role in the treatment of DN, which is probably attributable to the up - regulation of SIRT1 in renal inherent cells.