黄地安消胶囊调控磷脂酰基醇3-激酶/蛋白激酶B信号通路减轻GK大鼠血管病变损伤程度的研究

摘要

Objective To investigate the mechanisms of Huangdi Anxiao capsules in preventing and treating type 2 diabetes mellitus (T2DM) vascular lesions.Methods GK rats were randomly assigned into model group ,Shenqi Hypoglycemic Granules group ,Met-formin group,Huangdi Anxiao Capsules high,medium and low dosage groups ,and Wistar rats were chosen as the normal control groups , with 10 in each group.The model of T2DM vascular lesions was replicated by adding 0.1 g· L-1 Nω-nitro-L arginine methyl ester (L-NAME) to drinking water of GK rats and feeding with high fat diet .The model was administered with drugs at the same time ,once a day for 6 weeks.At the end of the experiment,the fasting blood glucose (FBG),serum total cholesterol (TC),triglyceride (TG),high-den-sity lipoprotein (HLDL),low-density lipoprotein (LDL) and oxidized low-density lipoprotein (ox-LDL) were measured.Hematoxylin-eosin staining (HE),Masson trichrome staining and Verhoeff staining were used to observe the degree of abdominal aortic vascular fi -brosis.The expressions of phosphatidylinositol 3-kinase (PI3K),phosphatase and tensin homolog deleted on chromosome ten (PTEN), tribbles gene 3 (TRIB3),Protein kinase B (PKB/Akt),hypoxia inducible factors (HIF-1α) and vascular endothelial growth factor (VEGF) mRNA in GK rats was detected by RT-qPCR.Results Compared with the normal group,the degree of pathological damage of the abdominal aorta,the levels of glucose and lipid metabolism ,PI3K,Akt,HIF-1αand VEGF mRNA expression in the model group were significantly increased (P <0.05 or P <0.01),and the expressions of PTEN,TRIB3 mRNA were significantly decreased (P <0.01).Compared with the model group ,the high dose (12.00 g· kg-1 ) and medium dose (6.00 g· kg-1 ) of Huangdi Anxiao cap-sules could not only improve the pathological injury of abdominal aorta in GK rats ,reduce the metabolism of glucose and lipid ,PI3K, Akt,HIF-1αand VEGF mRNA expression levels (P <0.05 or P <0.01),but also significantly increase PTEN,TRIB3 mRNA expres-sion levels (P <0.01).In the model group,the high dose and medium dose group of Huangdi Anxiao Capsules ,the expressions of PTEN mRNA were (0.42 ±0.22),(0.70 ±0.12),(0.55 ±0.08),the expressions of TRIB3 mRNA were (0.43 ±0.07),(0.75 ± 0.16),(0.55 ±0.04).Conclusions Huangdi Anxiao capsules has a certain therapeutic effect on T 2DM vascular lesions,the mecha-nism of which may be related to the regulation of PI3K/Akt signaling pathway and the improvement of glucose and lipid metabolism .%目的 探讨黄地安消胶囊治疗2型糖尿病(T2DM)血管病变的作用机制.方法 选取符合T2DM模型的GK大鼠按随机数字表法随机分为模型组、参芪降糖颗粒组、二甲双胍组、黄地安消胶囊高、中、低剂量组,另取Wistar大鼠为健康组,每组10只.通过在GK大鼠饮用水中加入0.1 g·L-1 Nω-硝基L精氨酸甲酯(L-NAME)联合高脂饲料喂养的方法复制T2DM血管病变大鼠模型.造模同时给药,每天1次,连续给药6周.实验结束后,测定大鼠的空腹血糖(FBG),血清生化指标总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白(HLDL)、低密度脂蛋白(LDL)、氧化低密度脂蛋白(ox-LDL);通过苏木精-伊红(HE)染色法、马松三色染色法、Verhoeff铁苏木精弹力纤维染色法观察腹主动脉血管纤维化程度;采用RT-qPCR技术检测GK大鼠腹主动脉中磷脂酰基醇3-激酶(PI3K)、抑癌基因(PTEN)、假性蛋白激酶3(TRIB3)、蛋白激酶B(PKB/Akt)缺氧诱导因子(HIF-1α)、血管内皮生长因子(VEGF)mRNA表达情况.结果 与健康组比较,模型组大鼠腹主动脉病理组织损伤程度明显增加,糖脂代谢水平、PI3K、Akt、HIF-1α、VEGF mRNA表达增加(P<0.05或P<0.01),PTEN、TRIB3 mRNA表达降低(P<0.01).与模型组比较,黄地安消胶囊高剂量组(12.00 g·kg-1)、中剂量组(6.00 g·kg-1)不仅可改善GK大鼠腹主动脉病理组织学损伤程度,明显降低糖脂代谢、PI3K、Akt、HIF-1α、VEGF mRNA表达水平(P<0.05或P<0.01),还可升高PTEN,TRIB3 mRNA表达水平(P<0.01),其中模型组、黄地安消胶囊高剂量组、中剂量组的PTEN mRNA表达水平分别为(0.42±0.22)、(0.70±0.12)、(0.55±0.08),TRIB3 mRNA表达水平分别为(0.43±0.07)、(0.75±0.16)、(0.55±0.04).结论 黄地安消胶囊对T2DM血管病变具有一定的治疗作用,其机制可能与调控PI3K/Akt信号通路、改善糖脂代谢有关.