葛根芩连汤通过IRS-1/PI3K/AKT通路对胃肠湿热型2型糖尿病大鼠糖脂代谢的影响

Effects of Gegenqinlian decoction on glycolipid metabolism of diabetes mellitus type 2 rats with pattern of dampness-heat in stomach and intestine through IRS-1/PI3K/AKT pathway

目的探究葛根芩连汤通过胰岛素受体底物-1(IRS-1)/磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路对胃肠湿热型2型糖尿病大鼠糖脂代谢的影响。方法将40只SD大鼠随机分为正常组(2 mL生理盐水灌胃)、造模组(2 mL生理盐水灌胃)、二甲双胍组(4.17 mg/100 g二甲双胍灌胃)和葛根芩连汤组(1 g/100 g葛根芩连汤灌胃),每组10只。采用高脂高糖饲料加腹腔注射链脲佐菌素(STZ)构建2型糖尿病大鼠模型,随后喂食油脂、42°白酒及蜂蜜水构建胃肠湿热型2型糖尿病大鼠模型。测量各组大鼠不同时间节点体质量,血糖仪测定空腹血糖(FBG);ELISA检测空腹胰岛素(FINS)、三酰甘油(TG)、总胆固醇(TC)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平变化、计算胰岛素抵抗指数(HOMA-IR);HE染色检测肝组织病理学变化;检测肝组织过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)及丙二醛(MDA)含量变化。Western blot检测肝组织IRS-1、PI3K、p-PI3K、AKT及p-AKT蛋白变化。结果与正常组比较,造模组大鼠体质量、FBG、FINS及HOMA-IR、GSH-Px、CAT、SOD、IRS-1、p-PI3K/PI3K及p-AKT/AKT水平均明显下降(P<0.05)、TG、TC、IL-6、TNF-α及MDA含量均显著升高(P<0.05),可见局灶性肝实质损失。与造模组比较,二甲双胍组及葛根芩连汤组大鼠体质量、FBG、FINS及HOMA-IR、GSH-Px、CAT、SOD、IRS-1、p-PI3K/PI3K及p-AKT/AKT水平均明显升高(P<0.05)、TG、TC、IL-6、TNF-α及MDA含量均显著降低(P<0.05),显示正常的肝实质。结论葛根芩连汤可明显改善胃肠湿热型2型糖尿病糖脂紊乱,可能是通过IRS-1/PI3K/AKT通路发挥作用。

Objective To explore the effects of Gegenqinlian decoction on glycolipid metabolism of diabetes mellitus type 2 rats with pattern of dampness-heat in stomach and intestine through insulin receptor substrate-1(IRS-1)/phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)pathway.Methods 40 SD rats were randomly divided into the normal group(2mL of normal saline by gavage),the modeling group(2mL of normal saline by gavage),the metformin group(4.17mg/100g metformin by gavage),and the Gegenqinlian decoction group(1g/100g Gegenqinlian decoction by gavage),with 10 rats in each group.The diabetes mellitus type 2 rat model was established with high-fat and high-sugar diet and intrabitoneal injection of streptozotocin(STZ),and then diabetes mellitus type 2 rat model with pattern of dampness-heat in stomach and intestine was established by feeding oil,42°white wine and honey water.The body weight of each group was measured at different time points,and the level of fasting blood glucose(FBG)was measured by glucose meter.The levels of fasting insulin(FINS),triglyceride(TG),total cholesterol(TC),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by ELISA,and insulin resistance index(HOMA-IR)was calculated.The histopathological changes of liver were detected by HE staining.The changes of catalase(CAT),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and malondialdehyde(MDA)levels in liver tissue were detected.The changes of IRS-1,PI3K,p-PI3K,AKT and p-AKT protein contents in liver tissue were detected by Western blot.Results Compared with the normal group,body weight,and levels of FBG,FINS,HOMA-IR,GSH-Px,CAT,SOD,IRS-1,p-PI3K/PI3K and p-AKT/AKT in the modeling group were significantly decreased(P<0.05),the levels of TG,TC,IL-6,TNF-αand MDA in the modeling group were significantly increased(P<0.05),indicating focal hepatic parenchymal loss.Compared with the modeling group,body weight,and the levels of FBG,FINS,HOMA-IR,GSH-Px,CAT,SOD,IRS-1,p-PI3K/PI3K and p-AKT/AKT in the metformin group and Gegenqinlian decoction group were significantly increased(P<0.05),while the levels of TG,TC,IL-6,TNF-αand MDA in the metformin group and Gegenqinlian decoction group were significantly decreased(P<0.05),indicating normal hepatic parenchyma.Conclusion Gegenqinlian decoction can significantly improve the glucolipid disorder of diabetes mellitus type 2 rats with pattern of dampnessheat in stomach and intestine,which may play the role through IRS-1/PI3K/AKT pathway.