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目的:探讨金匮肾气丸(Jinkui Shenqi Wan,JKSQ)对糖尿病大鼠骨骼肌胰岛素受体(insulin receptor,InsR)基因表达的影响,阐明其降低血糖的分子机制。方法:6周龄的SPF级雄性SD大鼠高脂高糖饲料喂养4周后,腹腔注射链脲佐菌素制备糖尿病模型。4 d后模型大鼠随机分成模型组、二甲双胍组、JKSQ低、高剂量组。正常组与模型组给予生理盐水,二甲双胍组给予二甲双胍(100 mg·kg-1),JKSQ低、高剂量组分别给予JKSQ(10,20 g·kg-1)灌胃4周。以酶学方法检测各组大鼠骨骼肌的己糖激酶(hexokinase,HK),6-磷酸果糖激酶-1(6-phosphofructokinase-1,PFK)活性,逆转录实时荧光定量PCR及蛋白印迹法检测骨骼肌InsR mRNA及蛋白表达水平变化。结果:与正常组比较,模型组出现了HK,PFK活性下降(P<0.01),InsR mRNA及蛋白质表达下降(P<0.01);给药治疗4周后,与模型组相比较,二甲双胍组、JKSQ低、高剂量组HK,PFK活性明显上升(P<0.05);同时各组InsR mRNA及蛋白质表达量明显提高(P<0.05)。结论:JKSQ能上调2型糖尿病大鼠骨骼肌InsR基因在mRNA及蛋白质的表达,增加HK与PFK活性,促进葡萄糖的氧化利用。
Objective: To investigate the effect of Jinkui Shenqi Wan (JKSQ) on the gene expression of insulin receptor (InsR) in skeletal muscle of diabetic rats and to elucidate its molecular mechanism of lowering blood glucose. Methods: Six-week-old SPF male SD rats were fed with high-fat and high-sugar diet for 4 weeks, then diabetic model was induced by intraperitoneal injection of streptozotocin. After 4 days, the model rats were randomly divided into model group, metformin group, JKSQ low and high dose groups. Rats in normal group and model group were given normal saline. Metformin group was given metformin (100 mg · kg-1), JKSQ low and high dose group were given JKSQ (10,20 g · kg-1) for 4 weeks. The activities of hexokinase (HK) and 6-phosphofructokinase-1 (PFK) in skeletal muscle of rats in each group were detected by enzymology method, and the mRNA and protein were detected by RT-PCR and Western blotting Changes of InsR mRNA and protein expression in skeletal muscle. Results: Compared with the normal group, the activity of HK and PFK in the model group decreased (P <0.01) and the expression of InsR mRNA and protein decreased (P <0.01). Compared with the model group, the metformin group, The activities of HK and PFK in JKSQ low and high dose groups were significantly increased (P <0.05), while the expression of InsR mRNA and protein in each group were significantly increased (P <0.05). Conclusion: JKSQ can up-regulate the mRNA and protein expression of InsR gene in skeletal muscle of type 2 diabetic rats, increase the activity of HK and PFK, and promote the oxidative utilization of glucose.