黄地安消胶囊调控PI3K/Akt通路改善自发性2型糖尿病大鼠血管病变损伤

Huang Di An Xiao capsules Regulates PI3K/Akt Pathway and Improves Vascular Lesion in Spontaneously Type 2 Diabetic Rats

目的:探讨黄地安消胶囊调控PI3K/Akt通路改善自发性2型糖尿病大鼠血管病变损伤的作用机制。方法:选取符合2型糖尿病大鼠模型的GK大鼠随机分为模型组、参芪降糖1. 08g/kg组、二甲双胍0. 72g/kg组、黄地安消胶囊12. 0、6. 0、3. 0g/kg组,另设Wistar大鼠为正常组,每组10只。通过在GK大鼠的饮用水中加入0. 1mg/ml Nω-精氨酸甲酯(L-NAME)联合高脂饲料喂养的方法复制2型糖尿病大血管病变的模型。造模同时开始给药,每日1次,连续6周。实验结束后,HE、Masson、Verhoeff染色法观察腹主动脉组织病理学损伤程度;免疫吸附法测定大鼠腹主动脉ET-1、HIF-1α和VEGF含量;免疫组织化学染色法观察腹主动脉I型胶原(Collagen I)、III型胶原(Collagen III)分布情况; Western blot技术检测大鼠腹主动脉内的TRIB3、PTEN、PI3K、Akt、p-PI3K、pAkt、HIF-1α、VEGF的蛋白表达水平。结果:与正常组比较,模型组大鼠腹主动脉中膜的厚度、胶原含量增加,弹力纤维减少,病理损伤程度明显增加,ET-1、HIF-1α、VEGF、Collagen I和Collagen III水平以及PI3K、Akt、p-PI3K、p-Akt、HIF-1α、VEGF蛋白表达显著增加,PTEN、TRIB3蛋白表达明显降低;与模型组比较,黄地安消胶囊不仅可改善GK大鼠腹主动脉组织病理学损伤程度,明显降低ET-1、HIF-1α、VEGF、Collagen I和Collagen III水平以及PI3K、Akt、p-PI3K、p-Akt、HIF-1α、VEGF蛋白表达水平,还可显著升高PTEN、TRIB3蛋白表达水平。结论:黄地安消胶囊能够改善自发性2型糖尿病大鼠血管病变损伤,其机制可能与调控PI3K/Akt信号通路、抑制腹主动脉胶原蛋白的沉积有关。

Objective： To explore the mechanism of action of Huang Di An Xiao capsules in regulating PI3 K/Akt pathway to improve vascular lesion in spontaneously type 2 diabetic rats. Methods： GK rats were selected and randomly divided into model group,Shen Qi Jiang Tang group,metformin group,Huang Di An Xiao capsules high,medium and low dose group,another Wistar rats were set as normal group,10 rats in each group. A model of macrovascular disease in type 2 diabetes mellitus was replicated by feeding High fat diet and 0. 1 mg/ml Nω-arginine methyl ester（L-NAME） was added to the drinking water of GK rats. At the same time of modeling,the drug are administered,once daily,for 6 weeks. The pathological damage of abdominal aorta was observed by HE,Masson and Verhoeff staining; the levels of ET-1,HIF-1α and VEGF were measured by immunoadsorption assay; immunohistochemical staining was used to observe the distribution of Collagen I and Collagen III in abdominal aorta,the protein expression levels of TRIB3,PTEN,PI3 K,Akt,p-PI3 K,p-Akt,HIF-1α and VEGF in rat abdominal aorta were detected by Western blot. Results： Compared with the normal group,the medial thickness of the abdominal aorta,collagen content increased,the elastic fibers decreased,the degree of pathological injury increased obviously in model group; the levels of ET-1,HIF-1α,VEGF,Collagen I and Collagen III and the protein expressions of PI3 K,Akt,p-PI3 K,p-Akt,HIF-1α and VEGF were significantly increased（P 〈 0. 05 or P 〈 0. 01）; PTEN,TRIB3 protein expression was significantly lower（P 〈 0. 01）; compared with model group,Huang Di An Xiao capsules could improve the histopathological damage of abdominal aorta in GK rats,decrease the levels of ET-1,HIF-1α,VEGF,Collagen I and Collagen III as well as PI3 K,Akt,p-Akt,HIF-1α,VEGF（P 〈 0. 05 or P 〈 0. 01）,and also significantly increased the expression of PTEN and TRIB3 protein（P 〈 0. 01）. Conclusion： Huang Di An Xiao capsules can improve spontaneous vascular injury in type 2 diabetic rats,the mechanism may be related to the regulation of PI3 K/Akt signaling pathway and inhibition of collagen deposition in the abdominal aorta.