糖肾平胶囊对糖尿病肾病KKAy小鼠肾脏保护作用及对Wnt/β-catenin信号通路的影响

Renal Protective Effect of Tangshiping Capsule on Diabetic Nephropathy Kkay Mice and Its Effect on Wnt/β-catenin Signal Transduction Pathway

目的:探讨糖肾平胶囊(糖肾平)对糖尿病肾病(DN) KKAy小鼠肾脏保护作用及其对Wnt/β-连环蛋白(β-catenin)途径信号通路的影响。方法:雌性10周龄SPF级KKAy小鼠60只,用KKAy鼠料诱导10周,以血糖>16. 7 mmol·L-1,24 h尿蛋白> 0. 4 mg确定DN小鼠模型成功。将成模小鼠随机分为模型组、厄贝沙坦组、糖肾平小、中、大剂量(0. 525,1. 05,2. 1 g·kg-1)组,10只雌性C57BL/6J小鼠作为正常组。每个治疗组均相应的治疗药物进行灌胃,正常组、模型组小鼠给予等体积的去离子水灌胃,灌胃剂量按0. 01 mL·g-1体质量系数,每天1次。观察小鼠一般情况,每4周称体质量并测24 h尿蛋白定量。第26周,眼球取血并处死小鼠,并测量肾脏的质量,分离血清测中血尿素氮(BUN),血肌酐(SCr),甘油三酯(TG),丙二醛(MDA),一氧化氮(NO)和超氧化物歧化酶(SOD)的含量;用原位杂交、免疫组化检测肾组织Wnt4,糖原合成酶激酶3β(GSK3β),β-连环蛋白(β-catenin) mRNA及蛋白的表达。结果:与模型组比较,糖肾平高剂量组体质量、肾重/体质量,24 h尿蛋白显著降低(P <0. 01);肾脏病理损害明显减轻,血清BUN,SCr,TG和MDA含量显著降低(P <0. 01),NO,SOD含量显著升高(P <0. 01),Wnt4,Gsk-3β,β-catenin mRNA及蛋白表达显著减少(P <0. 01)。结论:糖肾平可能是通过抑制Wnt/β-catenin信号通路活化,逆转DN KKAy小鼠肾小管上皮细胞转分化,延缓肾间质纤维化的进程,进而发挥肾脏保护作用。

Objective:To investigate the renal protective effect of Tangshenping capsule(Tangshenping on diabetic nephropathy(DN)KKAy mice and its effect on Wnt/β-catenin signaling pathway.Method:Sixty female Sprague-Dawley KKAy mice aged 10 weeks old were induced with KKAy rat feed for 10 weeks.The DN animal model was successfully determined with blood glucose(>16.7 mmol·L-1)and 24 hour urine protein(>0.4 mg).The model mice were randomly divided into a model group,an irbesartan group,and low,medium and high-dose Tangshenping group,with 10 female C57 BL/6 J mice as a control group.The treatment groups were given the corresponding drugs by gavage.The normal group and the model group were given an equal volume of deionized water by gavage.The intragastric dose was 0.01 m L·g-1 body weight coefficient once a day.The general conditions of the mice were observed,the body mass was weighed every 4 weeks,and 24 h urine protein was quantified.At the 26 thweek,the blood was collected from eyeballs,and the mice were put to death.The quality of the kidneys,serum blood urea nitrogen(BUN),serum creatinine(SCr),triglyceride(TG),malondialdehyde(MDA),nitric oxide(NO)and superoxide dismutase(SOD)content were measured.In situ hybridization and immunohistochemistry were used to detect the expressions of Wnt4,glycogen synthase kinase 3β(GSK3β)andβ-catenin in kidney tissues.Result:Compared with model group,body mass,kidney mass/body mass,and 24 h urine protein were significantly lower in high-dose Tangshenping group(P<0.01).The pathological damage of the kidney was significantly reduced,the serum BUN,SCr,TG and MDA contents were significantly decreased(P<0.01),the contents of NO and SOD were significantly increased(P<0.01),and the protein and mRNA expression levels of Wnt4,Gsk-3βandβ-catenin were decreased(P<0.01).Conclusion:Tangshenping may inhibit the activation of Wnt/β-catenin signaling pathway,reverse the transdifferentiation of renal tubular epithelial cells in DN KKAy mice,delay the progression of renal interstitial fibrosis,and then exert renal protection.