摘要
目的观察施今墨对药配方混悬液灌胃对2型糖尿病大鼠肝细胞胰岛素信号转导通路PI3K/AKT/GSK-3β的影响,探讨该方改善肝胰岛素抵抗的分子机制。方法40只雄性Wistar大鼠,随机抽取6只作为正常对照组,给予普通饲料喂养;余34只采用高脂饮食、高糖饮水联合腹腔注射小剂量链脲佐菌素建立2型糖尿病模型。取成模大鼠24只,随机分为模型对照组和施今墨对药配方低、中、高剂量组,每组各6只。正常对照组和模型对照组均给予1.5 mL/100 g纯净水灌胃,施今墨对药配方低、中、高剂量组分别给予该方混悬液(折合生药剂量5.8 g/kg、11.6 g/kg、23.2 g/kg)灌胃。葡萄糖氧化酶法测量大鼠血糖;ELISA法检测血清胰岛素水平,利用HOMA-IR公式计算胰岛素抵抗指数(IRI);糖原测定试剂盒测量肝糖原含量;Western blotting法检测PI3K、AKT、GSK-3β总蛋白表达及其磷酸化水平。结果与正常对照组比较,模型对照组血糖水平升高,IRI增加,肝糖原含量减少,PI3K、AKT蛋白及其磷酸化水平均降低,GSK-3β蛋白表达增加、磷酸化水平降低(P均<0.05)。与模型对照组比较,施今墨对药配方低剂量组血糖水平降低,IRI降低,肝糖原含量增加,PI3K与AKT蛋白及其磷酸化水平均增加,GSK-3β蛋白表达降低、磷酸化水平增加(P均<0.05);施今墨对药配方中、高剂量组上述指标改善不明显。结论施今墨对药配方能够增强2型糖尿病大鼠肝组织PI3K/AKT/GSK-3β信号通路,从而增加肝细胞中的糖原含量,减轻胰岛素抵抗程度,降低血糖。
Objective To observe the effects of Shijinmo herbal-pair formula on the liver glycogen content,and PI3K/AKT/GSK-3βsignaling pathway in type 2 diabetic rats,and to explore its action mechanism in reducing the insulin resistance.Methods Six rats were randomly selected from 40 rats and given common diet,and the rest were given high-fat/sugar diet combined with intraperitoneal injection of low-dose streptozotocin to establish type 2 diabetic models.Twenty-four diabetic rats were randomly divided into the model control group,and low-dose,medium-dose and high-dose Shijinmo herbal-pair formula groups,with 6 rats in each group.The rats in the normal control group and the model control group were given pure water(1.5 mL/100 g),and the rats in the low-dose,medium-dose and high-dose Shijinmo herbal-pair formula groups were given the corresponding drugs(equals to the original herbs of 5.8,11.6 and 23.2 g/kg)for 8 weeks.Blood glucose was measured with glucose oxidase method.The level of serum insulin was measured by ELISA.The Homa-IR formula was used to calculate IR index.The hepatic glycogen content was measured by the glycogen assay kit;the protein expression levels of PI3K,AKT and GSK-3β,and their phosphorylation levels were analyzed by Western blotting.Results Compared with the normal control group,the blood glucose level increased,IRI increased,liver glycogen content decreased,the protein express levels of PI3K and AKT,and their phosphorylation levels decreased,GSK-3βprotein expression increased,but its phosphorylation level decreased in the model control group(all P<0.05).Compared with the model control group,the blood glucose level and IRI value were significantly reduced,the glycogen content in the liver increased in type 2 diabetic rats after the treatment of low-dose Shijinmo herbal-pair formula group.The low-dose Shijinmo herbal-pair formula increased the protein expression levels of PI3K and AKT and their phosphorylation levels,and decreased the protein level of GSK-3βbut enhanced its phosphorylation level(all P<0.05).The above indicators were not significantly improved in the medium-dose and high-dose Shijinmo herbal-pair formula groups.Conclusion Shijinmo herbal-pair formula has hypoglycemic effect through enhancing the insulin signal transduction pathway of PI3K-AKT-GSK-3β,thereby increasing the glycogen content in hepatocytes and reducing the degree of IR.
作者
代紫阳
董玉山
李继安
王亚
常宏
杜晨光
DAI Ziyang;DONG Yushan;LI Ji′an;WANG Ya;CHANG Hong;DU Chenguang(North China University of Science and Technology,Tangshan 063210,China)
出处
《山东医药》
CAS
2020年第14期35-38,共4页
Shandong Medical Journal
基金
河北省科技计划项目(17397733D)
华北理工大学研究生创新项目(2018S55)。
