基于AMPK探究电针“天枢”结合二甲双胍对2型糖尿病大鼠的降糖效应  

Hypoglycemic effect of electroacupuncture at "Tianshu"(ST 25) combined with metformin on rats with type 2 diabetes mellitus based on AMPK

作　　者：申学婷 张双双[1] 陈小燕 余芝[1] 徐斌[1] SHEN Xue-ting;ZHANG Shuang-shuang;CHEN Xiao-yan;YU Zhi;XU Bin(Key Laboratory of Acupuncture and Medicine Research of Ministry of Education,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu Province,China)

机构地区：[1]南京中医药大学针药结合教育部重点实验室,江苏南京210023

出　　处：《中国针灸》2023年第1期53-59,共7页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金资助项目:81873238、82074532、81673883;江苏省中医学优势学科三期项目立项开放课题项目:ZYX03KF012;江苏省中医药领军人才资助项目:SLT0225。

摘　　要：目的:观察电针“天枢”结合二甲双胍对2型糖尿病(T2DM)模型大鼠的降糖效应及对肝脏和胰腺组织腺苷酸活化蛋白激酶(AMPK)表达的影响。方法:将36只雄性SD大鼠随机分为空白组(6只)和造模组(30只)。造模组大鼠采用高脂饲料喂养配合小剂量链脲佐菌素(STZ)腹腔注射的方法建立T2DM模型,将造模成功的大鼠随机分为模型组、对照组、二甲双胍组、电针组和针药结合组,每组6只。电针组大鼠予电针“天枢”干预,疏密波,频率2 Hz/15 Hz,电流强度2 mA,每次20 min;二甲双胍组大鼠采用二甲双胍(190 mg/kg)溶于0.9%氯化钠溶液灌胃(2 mL/kg);针药结合组予电针“天枢”结合二甲双胍溶液灌胃;对照组大鼠予同等剂量0.9%氯化钠溶液灌胃。均每日1次,连续干预5周。干预结束后,检测大鼠体质量和随机血糖;ELISA法检测大鼠血清胰岛素水平;Western blot法检测大鼠肝脏和胰腺组织AMPK及磷酸化腺苷酸活化蛋白激酶(p-AMPK)蛋白表达;免疫荧光法检测大鼠胰腺组织蛋白基因产物9.5(PGP9.5)表达。结果:(1)与空白组比较,模型组体质量降低(P<0.05);与模型组比较,电针组及针药结合组体质量降低(P<0.05);电针组和针药结合组体质量低于二甲双胍组(P<0.05)。与空白组比较,模型组随机血糖升高(P<0.01);与模型组比较,二甲双胍组、电针组、针药结合组随机血糖降低(P<0.01);针药结合组随机血糖低于二甲双胍组和电针组(P<0.05)。(2)与空白组比较,模型组胰岛素水平降低(P<0.05);与模型组比较,二甲双胍组、电针组、针药结合组胰岛素水平均升高(P<0.05);针药结合组胰岛素水平高于二甲双胍组和电针组(P<0.05)。(3)与空白组比较,模型组肝脏组织AMPK及p-AMPK蛋白表达均减少(P<0.05),胰腺组织AMPK和p-AMPK蛋白表达增加(P<0.05)。与模型组比较,二甲双胍组、电针组、针药结合组肝脏组织AMPK和p-AMPK蛋白表达均增加(P<0.05,P<0.01);二甲双胍组Objective To observe the hypoglycemic effect of electroacupuncture(EA) at "Tianshu"(ST 25) combined with metformin on rats with type 2 diabetes mellitus(T2DM) as well as its effect on expression of adenosine monophosphate activated protein kinase(AMPK) in liver and pancreas. Methods Thirty-six male SD rats were randomly divided into a blank group(6 rats) and a model establishing group(30 rats). The rats in the model establishing group were fed with high-fat diet and treated with intraperitoneal injection of low-dose streptozotocin(STZ) to establish T2DM model. The rats with successful model establishment were randomly divided into a model group, a control group, a metformin group, an EA group and a combination group, 6 rats in each group. The rats in the EA group were treated with EA at "Tianshu"(ST 25),dense-disperse wave, 2 Hz/15 Hz in frequency and 2 mA in current intensity, 20 min each time. The rats in the metformin group were treated with intragastric administration of metformin(190 mg/kg) dissolved in 0.9% sodium chloride solution(2m L/kg). The rats in the combination group were treated with EA at "Tianshu"(ST 25) and intragastric administration of metformin. The rats in the control group were treated with intragastric administration of 0.9% sodium chloride solution with the same dose. All the treatments were given once a day for 5 weeks. After the intervention, the body mass and random blood glucose were detected;the serum insulin level was detected by ELISA;the expression of AMPK and phosphorylated adenosine monophosphate activated protein kinase(p-AMPK) in liver and pancreas was detected by Western blot method;the expression of protein gene product 9.5(PGP9.5) was detected by immunofluorescence. Results(1)Compared with the blank group, the body mass in the model group was decreased(P<0.05);compared with the model group, the body mass in the EA group and the combination group was decreased(P<0.05);the body mass in the EA group and the combination group was lower than the metformin group(P<0.05). Compared

关 键 词：2型糖尿病(T2DM) 电针 穴 天枢 二甲双胍 针药结合 腺苷酸活化蛋白激酶(AMPK) 

分 类 号：R245.97[医药卫生—针灸推拿学]