中药降糖复方水提物通过PI3K/Akt/NF-κB信号通路抑制KK-Ay小鼠糖尿病肾病炎症  

Inhibition of Water extract from Jiangtang Decoction on inflammation in KK-Ay mice by PI3K/Akt/NF-κB signaling pathway

作　　者：洪金妮[1] 黎巍威[1] 张宁[1] 富宏[1] 王学美[1] 

机构地区：[1]北京大学第一医院中西医结合研究室,北京100034

出　　处：《药物评价研究》2017年第10期1389-1396,共8页Drug Evaluation Research

基　　金：国家自然科学基金面上项目(81573763);国家自然科学基金重点项目(81530099);北京市自然科学基金资助项目(7172221);国家重点研发计划(2016YFE0116200)

摘　　要：目的应用自发性2型糖尿病模型KK-Ay小鼠,研究中药降糖复方水提物(WEJTD)对糖尿病及其引起的糖尿病肾病(DN)的疗效及作用机制。方法将KK-Ay小鼠按随机数法随机分为5组:模型组、盐酸二甲双胍(250 mg/kg)组和WEJTD低、中、高剂量(2、4、8 g/kg)组,C57BL/6J小鼠作为对照组,每天ig给药1次,共12周,对照组和模型组ig等体积蒸馏水。给药12周后,将小鼠放入代谢笼,检测摄食、饮水及尿量;内眦取血,ELISA法检测血清中白细胞介素-6(IL-6)、细胞间黏附分子-1(ICAM-1)、肿瘤坏死因子-α(TNF-α)含量;随后处死小鼠,取小鼠肾脏做苏木精-伊红(HE)及过碘酸六胺银(PASM)染色;取肾组织,采用蛋白免疫印迹(Western blotting)和实时荧光定量PCR(q RT-PCR)法检测磷脂酰肌醇激酶-3(PI3K)、蛋白激酶B(Akt)、核因子-κB(NF-κB)、IL-6、ICAM-1、TNF-α等指标。结果 WEJTD显著减少KK-Ay小鼠摄食、饮水和尿量(P<0.05、0.01、0.001);缓解小鼠肾脏的病理形态学变化,显著改善糖原沉积(P<0.001);显著降低血清和肾脏组织中炎症因子IL-6、ICAM-1和TNF-α水平(P<0.05、0.01、0.001);显著上调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)。结论 WEJTD显著改善KK-Ay小鼠糖尿病症状、肾脏病理变化,可能与激活PI3K-Akt信号通路,进而抑制NF-κB磷酸化,降低体内炎症因子有关。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. Resdts 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.

关 键 词：中药降糖复方 水提物 KK-AY小鼠 2型糖尿病 糖尿病肾病 炎症因子 PI3K/Akt/NF-κB 

分 类 号：R965.1[医药卫生—药理学]