糖肾宁对糖尿病肾病KKAy小鼠的肾脏保护作用及其对Notch/Snail1信号转导通路的影响

Renoprotective Effect of Tang-Shen-Ning and Its Influence on Notch/Snail1 Pathway in Diabetic Nephropathy KKAy Mice

目的:明确糖肾宁是否通过干预Notch/snail1通路抑制KKAy小鼠糖尿病肾病肾小管上皮细胞转分化肾间质纤维化。方法:KKAy小鼠30只,经过10周专用鼠料饲养,血糖>16.7 mmol·L-1,24 h尿蛋白>0.4 mg为糖尿病肾病动物模型。造模成功小鼠按血糖和体质量分层随机分为模型组、厄贝沙坦组与糖肾宁组,灌胃给药,雌性C57BL/6J小鼠10只作为正常对照组。检测小鼠一般状况,称体重、测24 h尿蛋白定量。干预16周后取材,检测血糖、尿素氮(Blood Urea Nitrogen,BUN)、血清肌酐(Serum creatinine,Scr);肾组织行HE、Mallory染色。原位杂交及Western blotting法检测小鼠肾组织中Notch/snail1通路、α-SMA、E-Cadherin蛋白及m RNA表达。SPSS20.0软件进行统计学处理。结果:与模型组比较,糖肾宁组小鼠一般状态改善,体质量、24 h尿蛋白定量显著减少(P<0.01),血清BUN及Scr含量降低(P<0.01,P<0.05),病理染色显示肾间质纤维化明显减轻,糖肾宁组肾组织Notch/snail1通路和α-SMA蛋白及m RNA表达明显降低,有显著统计学差异(P<0.01),E-Cad蛋白及m RNA表达显著升高(P<0.01)。结论:糖肾宁可以保护糖尿病肾病肾功能,延缓糖尿病肾病进展,抑制糖尿病肾病肾小管上皮细胞转分化肾间质纤维化,改善糖尿病肾病EMT的机制可能是通过抑制Notch/snail1通路表达实现。

This paper was aimed to study the renal protective effect of Tang-Shen-Ning （TSN） on diabetic nephropathy （DN） KKAy mice by inhibiting the Noteh/snaill signal transduction pathway. A total of 30 KKAy mice, which were fed with mice-dedicated food for 10 weeks and with the blood glucose over 16.7 mmol. L-1, 24-hour urinary albumin larger than 0.4 mg, were made into the DN model. The DN mice were randomly divided into the model group, irbesartan group and TSN group according to their blood glucose and weight. Intragastric administration of medication was given. A total of 10 female C57BL/6J mice were selected as the control group. The general condition, body weight and 24-hour urinary protein quantitation were detected. After 16-week intervention, mice were sacrificed. Levels of blood glucose, blood urea nitrogen （BUN） and serum creatinine （Scr） were detected. HE and Mallory staining were applied to renal tissues. In situ hybridization （ISH） and western blotting were used to detect the Notch/snaill pathway, α-SMA, E-Cadherin protein and mRNA expression in renal tissues. Statistical analysis was made by SPSS20.0 software. The results showed that compared with the model group, the rats＇ general conditions were improved; body weight and 24-hour urinary protein quantitation were significantly decreased （P〈0.01）; contents of BUN and Scr were reduced （P〈0.01, P〈0.05）. The pathological staining showed significantly reduction on renal interstitial fibrosis. The Notch/snaill pathway, protein and mRNA expression of α-SMA were significant reduced with statistical significance （P〈0.01）; protein and mRNA expression of E- Cad protein were significant increased with statistical significance （P〈0.01）. It was concluded that TSN can protect the renal function of DN, delay the disease progression of DN, and inhibit epithelial-mesenchymal transdifferentiation （EMT） of renal tubular epithelial cells and renal interstitial fibrosis. Furthermore, the inhibition on EMT may be through the regulation of the Notch/snail 1 pathway.