基于AGEs-RAGE信号通路研究中药复方益糖康改善大鼠骨骼肌胰岛素抵抗的作用机制

Research on Mechanism of Yitangkang(益糖康)Ameliorating Insulin Resistance in Skeletal Muscle of Rats Based on AGEs-RAGE Signal Pathway

目的研究益糖康改善大鼠骨骼肌胰岛素抵抗的效果及作用机制。方法8周龄健康雄性Wistar大鼠96只,分别设置空白对照组、模型对照组、益糖康高剂量组、益糖康中剂量组、益糖康低剂量组、西药对照组(盐酸吡格列酮),剂量分别为(40 g/kg、20 g/kg、10 g/kg、1.35 mg/kg),运用高糖高脂糖尿病模型诱导饲料联合小剂量链唑霉素(STZ)腹腔注射建立胰岛素抵抗动物模型并运用高胰岛素-正葡萄糖钳实验验证,模型判定成功后对各组大鼠按要求进行灌胃给药干预。用稳态胰岛素抵抗指数及胰岛素抗体水平评价给药干预后各组大鼠胰岛素抵抗的变化情况,运用免疫组化及免疫蛋白印迹法对AGEs-RAGE信号通路核心蛋白进行验证。结果胰岛素抵抗动物模型建立成功,药物干预后益糖康高剂量组、中剂量组和西药对照组的稳态胰岛素抵抗指数、胰岛素抗体水平显著降低(P<0.01)。免疫组化与免疫蛋白印迹法结果提示,与空白对照组相比,模型对照组骨骼肌组织的AGEs、RAGE、PKCβⅡ、Egr-1、JNK、JAK2、STAT1、STAT3的蛋白表达水平显著升高;与模型对照组相比,益糖康高剂量组、中剂量组和西药对照组骨骼肌组织中上述蛋白的表达水平显著降低(P<0.01)。结论益糖康可能通过抑制AGE-RAGE通路中的AGEs、RAGE、PKCβⅡ、Egr-1、JNK、JAK2、STAT1、STAT3蛋白的表达水平来发挥抗炎、抑制氧化应激和细胞凋亡的作用,进而改善胰岛素抵抗。

Objective To explore the effect and mechanism of Yitangkang(益糖康)ameliorating the insulin resistance in skel-etal muscle of rats.Methods A total of 968-week-old healthy male Wistar rats were divided into blank control group,model control group,Yitangkang high(40 g/kg),medium(20 g/kg)and low(10 g/kg)dose groups and western medicine control group(pioglitazone hydrochloride,1.35mg/kg)respectively.The animal model of insulin resistance was established by high glucose and high-fat diabetes model induction feed combined with low-dose streptozotocin(STZ)intraperitoneal injection,and verified by high insulin glucose clamp experiment.After the model was set up successfully,the rats in each group were given intragastric administration intervention as required.The homeostasis insulin resistance index and insulin antibody level were used to evaluate the changes of insulin resistance in each group after the intervention.The core proteins of AGEs-RAGE signal pathway were validated by the technique of immunohistochemistry and Western blot.Results The results pointed out that the animal model of insulin resistance was successfully established.After intervention with the above-mentioned drugs,the homeostasis insulin re-sistance index and insulin antibody level of the Yitangkang high and medium dose groups and western medicine control group were significantly decreased(P<0.01).The results of immunohistochemical and Western blotting pointed out that compared with those of the blank control group,the protein expression levels of advanced glycation end products(AGEs),receptor for advanced glycation end products(RAGE),protein kinase CβⅡ(PKCβⅡ),early growth responsive gene-1(Egr-1),c-Jun N-termi-nal kinase(JNK),Janus kinase 2(JAK2),signal transducer and activator of transcription 1(STAT1)and signal transducer and activator of transcription 3(STAT3)in skeletal muscle tissue of rats in the model control group were increased significantly.Compared with those of the model control group,the above-mentioned protein expression levels in the Yitangkang high and me-dium dose groups and western medicine control group were decreased significantly(P<0.01).Conclusion Yitangkang may in-hibit the protein expression levels of AGEs,RAGE,PKCβⅡ,Egr-1,JNK,JAK2,STAT1 and STAT3 to play the role of anti-in-flammatory,inhibiting oxidative stress and apoptosis,so as to improve the insulin resistance.