马欢,高楠楠,陈俞如,肖瑛,潘艳伶
MA Huan,GAO Nannan,CHEN Yuru,XIAO Ying,PAN Yanling

• 1:贵州医科大学
• 2:贵州省常见慢性疾病发病机制及药物研究重点实验室
• 3:贵州医科大学附院

摘要(Abstract)：

目的:观察糖通饮对2型糖尿病(T2DM)大鼠胰腺组织磷脂酰肌醇3激酶(PI3K)-蛋白激酶B(AKT)信号通路的影响,探讨糖通饮对2型糖尿病大鼠胰腺组织的保护作用及其机制。方法:50只SPF级SD雄性大鼠,随机均分为对照组,模型组,糖通饮低、中及高剂量组;对照组给予正常饲料喂养,其余4组大鼠采用高脂饲料联合小剂量多次腹腔注射链脲佐菌素制备T2DM大鼠模型,造模成功的糖通饮低、中及高剂量组大鼠给予糖通饮(10、20及30 mg/kg)灌胃治疗8周,对照组和模型组大鼠给予同体积双蒸水处理;比较各组大鼠给药前,干预2、4及8周时的体质量,比较干预前及干预8周时各组大鼠的空腹血糖(FBG);干预8周时,逆转录实时荧光定量PCR法检测大鼠胰腺组织中AKT及胰岛素受体底物1(IRS-1) mRNA水平,免疫组织化学法检测大鼠胰腺组织中AKT及IRS-1蛋白表达。结果:干预前,干预2、4及8周时,模型组大鼠体质量较对照组显著降低(P<0.01);干预2周和4周时,糖通饮高剂量组大鼠体质量较模型组显著升高(P<0.05);干预8周时,糖通饮各剂量组大鼠体质量均较模型组显著升高(P<0.01),但各剂量组间比较,差异无统计学意义(P>0.05);干预8周时,与对照组比较,模型组大鼠FBG显著升高(P<0.01),胰腺组织AKT及IRS-1 mRNA和蛋白的表达均明显降低(P<0.01);与模型组比较,糖通饮各剂量组大鼠FBG明显降低(P<0.01),糖通饮高剂量组AKT及IRS-1 mRNA的表达水平显著升高(P<0.01),糖通饮高剂量组AKT和各剂量组IRS-1蛋白的表达也均显著增加(P<0.01)。结论:糖通饮可通过激活胰腺组织PI3K/AKT信号通路,改善T2DM大鼠胰岛素抵抗和降低血糖,其机制可能与影响PI3K/AKT信号通路中因子AKT的表达水平有关。
Objective: To observe the effect of Tangtongyin formula on PI3 K-AKT pathway in pancreas tissues of rats with Type 2 diabetes mellitus(T2 DM), and to explore the protective effect and mechanism of Tangtongyin formula on pancreas tissues of rats with T2 DM. Methods: 50 SPF SD male rats were randomly divided into the control group, the model group and the low-dose, medium-dose and high-dose Tangtongyin groups, with 10 rats in each group. The control group took normal feed, and the other 4 groups with high fat feed combined with small dose of urea with intraperitoneal injection of multiple chain rhzomorph preparation of T2 DM rat model; Successful medel groups with Tangtongyin formula low, medium and high doses received gastric gavage of Tangtongyin formula(10, 20 and 30 mg/kg respectively) for 8 weeks; The control group and model group were given with double steaming water volume, and the comparison of body quality among the groups before intervention, and of 2-, 4-and 8-week intervention, and the comparison of fasting blood glucose(FBG), mRNA levels of AKT and insulin receptor substrate 1(IRS-1) in the rat pancreas were made by real-time fluorescence quantitative PCR intervention for 8 weeks; The protein expressions of AKT and IRS-1 in the rat pancreas after intervention for 8 weeks were detected by immunohistochemistry. After 8 weeks, body weight, FBG were measured. The expression of AKT and IRS-1 in rat pancreas was detected by real-time fluorescence quantitative PCR, and the expression of AKT and IRS-1 protein in rat pancreas was detected by immunohistochemistry. Results: Before intervention, and after intervention for 2, 4 and 8 weeks the body mass of rats in the model group was significantly lower than that in the control group(P<0.01). After intervention for 2 weeks and 4 weeks, the body mass of rats in the high-dose group was significantly higher than that in the model group(P<0.05). After intervention for 8 weeks, the body mass of rats in each Tangtongyin dose group was significantly higher than that in the model group(P<0.01), but the difference was not statistically significant(P>0.05). After 8 weeks' intervention, FBG in the model group significantly increased more than in the control group(P<0.01), and the mRNA and protein expressions of AKT and IRS-1 in the pancreatic tissues decreased significantly(P<0.01). Compared with the model group, FBG decreased significantly in rats with all Tangtongyin doses(P<0.01), the expression levels of AKT and IRS-1 mRNA increased significantly in the high-dose Tangtongyin group(P<0.01), and the expression levels of AKT and IRS-1 protein increased significantly in the high-dose Tangtong group(P<0.01).Conclusion: Tangtongyin formula can improve insulin resistance and reduce blood glucose in T2 DM rats by activating the PI3 K/AKT signaling pathway in the pancreatic tissues, and its mechanism may be related to the expression level of factor AKT in the PI3 K/AKT signaling pathway.

关键词(KeyWords)： 2型糖尿病;糖通饮;胰腺组织;糖代谢;胰岛素受体底物1;磷脂酰肌醇3激酶;蛋白激酶B
Type 2 diabetes mellitus;Tangtongyin formula;pancreatic tissue;glucose metabolism;insulin receptor substrate 1;phosphatidylinositol 3-kinase;protein kinase B;insulin resistance

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技计划项目课题[黔科合LH字(2016)7248号];; 贵州省中医药管理局中医药、民族医药科学技术研究课题(QZYY-2016-006)

作者(Author): 马欢,高楠楠,陈俞如,肖瑛,潘艳伶
MA Huan,GAO Nannan,CHEN Yuru,XIAO Ying,PAN Yanling

DOI: 10.19367/j.cnki.1000-2707.2020.01.010

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