消渴饮水方对链脲佐菌素联合高脂高糖诱导的糖尿病肾病小鼠的保护作用

目的：研究消渴饮水方（XEC）对实验性2型糖尿病（T2DM）小鼠糖脂代谢及肾病损伤的保护作用及机制。方法：采用链脲佐菌素联合高脂高糖饲料诱导C57BL/6小鼠为T2DM动物模型，分为模型（DM）组、二甲双胍（MET）组、消渴灵片（XKLP）组、消渴饮水方低（XEC-L）、中（XEC-M）、高（XEC-H）剂量组，并以健康小鼠为正常（NC）组，给药6周。每周测空腹血糖，第6周进行口服葡萄糖耐量实验，检测糖化血红蛋白、空腹胰岛素、尿素氮等生化指标；HE染色观察胰腺病理改变，HE、PAS和Masson染色观察肾脏病理改变；蛋白质印迹（Western blot）检测肝脏组织胰岛素受体α（InsRα）、胰岛素受体底物1（IRS-1）、磷酸脂酰激醇-3激酶（PI3K）、蛋白激酶B （AKT）、腺苷酸化蛋白激酶（AMPK）和磷酸化AMPK （p-AMPK）水平，肾脏组织晚期糖基化终末产物受体（RAGE）、核因子-κB （NF-κB）、c-Jun氨基末端激酶（JNK）和磷酸化JNK （p-JNK）水平。结果：XEC显著改善T2DM小鼠的胰岛素抵抗、糖耐量降低和糖脂代谢紊乱（P<0.01），以及受损的肾功能（P<0.05）。在组织病理分析中，XEC减轻T2DM小鼠胰腺、肾小球和肾小管的结构病变。Western blot分析表明，XEC-H显著上调InsRα、IRS-1、PI3K、AKT的表达和p-AMPK/AMPK比值，降低RAGE、NF-κB的水平和p-JNK/JNK比值（P<0.05）。结论：XEC能够改善T2DM小鼠的糖脂代谢紊乱，在预防治疗糖尿病肾病方面显示出比MET和XKLP更优异的效果。其作用机制可能是通过上调InsRα/IRS-1/PI3K/AKT信号通路，促进AMPK磷酸化及下调肾脏AGE/RAGE和JNK/NF-κB信号通路，实现对糖尿病及肾病的预防和治疗作用。

Protective effect of Xiaoke Yinshui extract combination on HSHF diet/Streptozotocin-Induced diabetes and diabetic nephropathy in mice

OBJECTIVE To investigate the protective effect and mechanism of Xiaoke Yinshui extract combination (XEC) on glycolipid metabolism and kidney injury in type 2 diabetes mellitus (T2DM) mice.METHODS Mouse model of T2DM were established by feeding C57BL/6 mice with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin.The T2DM mice were divided into model group (DM), metformin group (MET), Xiaokeling Pian group (XKLP), low-, medium-, high-dose of XEC group (XEC-L, XEC-M, XEC-H) and other normal mice were treated as normal group (NC).Mice were orally administered every day for 6 weeks and fasting blood glucose was examined every week.The oral glucose tolerance tests were performed and biochemical parameters were measured in each group of mice.HE staining was used to observe pathological changes of pancreas and the changes of renal tissues were observed by HE, PAS and Masson staining.The expressions of InsRα, IRS-1, PI3K, AKT, AMPK and p-AMPK in liver tissue and RAGE, NF-κB, JNK and p-JNK in renal tissue were detected by Western blot.RESULTS Insulin resistance, impaired glucose tolerance and glycolipid metabolism disorder were improved significantly by XEC (P<0.01).And indicators related to renal function, oxidative stress, inflammation and fibrosis were also ameliorated (P<0.05).Compared with DM and XKLP group, pathological changes of pancreas and renal were alleviated in XEC groups.The expressions of InsRα, IRS-1, PI3K, AKT, p-AMPK/AMPK ratio were significantly up-regulated in XEC-H group.Meanwhile, the level of RAGE, NF-κB and p-JNK/JNK ratio were down-regulated (P<0.05).CONCLUSION XEC can improve the glycolipid metabolism disorder in T2DM mice and shows a better effect than MET and XKLP in the treatment of diabetic nephropathy.This effect may be related to the up-regulation of InsRα/IRS-1/PI3K/AKT signaling pathway and phosphorylation of AMPK, and the down-regulation of AGE/RAGE and JNK/NF-κB signaling pathway.