目的 應(yīng)用自發(fā)性2型糖尿病模型KK-Ay小鼠，研究中藥降糖復(fù)方水提物（WEJTD）對糖尿病及其引起的糖尿病腎病（DN）的療效及作用機制。方法 將KK-Ay小鼠按隨機數(shù)法隨機分為5組：模型組、鹽酸二甲雙胍（250 mg/kg）組和WEJTD低、中、高劑量（2、4、8 g/kg）組，C57BL/6J小鼠作為對照組，每天ig給藥1次，共12周，對照組和模型組ig等體積蒸餾水。給藥12周后，將小鼠放入代謝籠，檢測攝食、飲水及尿量；內(nèi)眥取血，ELISA法檢測血清中白細胞介素-6（IL-6）、細胞間黏附分子-1（ICAM-1）、腫瘤壞死因子-α（TNF-α）含量；隨后處死小鼠，取小鼠腎臟做蘇木精-伊紅（HE）及過碘酸六胺銀（PASM）染色；取腎組織，采用蛋白免疫印跡（Western blotting）和實時熒光定量PCR（qRT-PCR）法檢測磷脂酰肌醇激酶-3（PI3K）、蛋白激酶B（Akt）、核因子-κB（NF-κB）、IL-6、ICAM-1、TNF-α等指標。結(jié)果 WEJTD顯著減少KK-Ay小鼠攝食、飲水和尿量（P<0.05、0.01、0.001）；緩解小鼠腎臟的病理形態(tài)學(xué)變化，顯著改善糖原沉積（P<0.001）；顯著降低血清和腎臟組織中炎癥因子IL-6、ICAM-1和TNF-α水平（P<0.05、0.01、0.001）；顯著上調(diào)PI3K和Akt的磷酸化水平（P<0.05、0.01、0.001）；抑制NF-κB磷酸化水平（P<0.001）。結(jié)論 WEJTD顯著改善KK-Ay小鼠糖尿病癥狀、腎臟病理變化，可能與激活PI3K-Akt信號通路，進而抑制NF-κB磷酸化，降低體內(nèi)炎癥因子有關(guān)。

Objective To study the effect and mechanism of Water extract from Jiangtang Decoction (WEJTD) on diabetes mellitus and diabetic nephropathy (DN) in spontaneous type 2 diabetes mellitus model KK-Ay mice. Methods Totally 50 KK-Ay mice were randomly divided into five groups:model group, metformin (positive drug, 250 mg/kg) group, WEJTD low, medium and high dose (2, 4, and 8 g/kg) group, with 10 C57BL/6J mice as normal group. The relative drugs were ig administered once a day for 12 weeks, and mice in control group and model group were perfused with distilled water of equal volume. After 12 weeks' oral administration, mice were put into metabolism cages, and the food-intake, water-intake and urine volume were calculated and collected. Blood were collected to detect the concentration of IL-6, ICAM-1 and TNF-α. Then mice were executed, and HE staining and PASM staining were used to check the effect of WEJTD on kidney. Western blotting and qRT-PCR were used to detect the concentration of PI3K, Akt, NF-κB, IL-6, ICAM-1 and TNF-α in kidney. Results WEJTD can alleviate the symptoms of diabetes, such as food ration, polydipsia and polyuria (P<0.05, 0.01, and 0.001); Relief the pathological changes of kidney and significantly decreased glycogen deposition (P<0.001), down-regulate the increase of IL-6, ICAM-1 and TNF-α in serum and kidney (P<0.05, 0.01 and 0.001), up-regulate the phosphorylation of PI3K and Akt (P<0.05, 0.01, and 0.001), and inhibit the phosphorylation of NF-κB (P<0.001). Conclusion WEJTD had positive effects on kidney morphology of KK-Ay, and the underlying mechanism might be related to the regulation of PI3K-Akt and NF-κB- mediated inflammation.

國家自然科學(xué)基金面上項目（81573763）；國家自然科學(xué)基金重點項目（81530099）；北京市自然科學(xué)基金資助項目（7172221）；國家重點研發(fā)計劃（2016YFE0116200）