电针对糖尿病胃轻瘫大鼠胃Cajal间质细胞自噬信号通路的影响  

Effects of electroacupuncture on autophagy signaling pathway of gastric antrum interstitial cells of Cajal in diabetic gastroparesis rats

作　　者：张天华 魏星 黎晓宇 肖小娟 赵莎彤 赵东凤 钟艳 彭艳[1] ZHANG Tianhua;WEI Xing;LI Xiaoyu;XIAO Xiaojuan;ZHAO Shatong;ZHAO Dongfeng;ZHONG Yan;PENG Yan(College of Acupuncture-moxibustion-Tuina and Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208,China;School of Informatics,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学针灸推拿与康复学院,湖南长沙410208 [2]湖南中医药大学信息科学与工程学院,湖南长沙410208

出　　处：《中国病理生理杂志》2023年第4期639-646,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81774431);湖南省自然科学基金资助项目(No.2022JJ40310);湖南省教育厅科学研究项目(No.22A0275);长沙市自然科学基金资助项目(No.kq2202263);湖南省研究生科研创新项目(No.2022CX98);湖南中医药大学校级重点项目(No.2022XJZKA001)。

摘　　要：目的:探讨电针对糖尿病胃轻瘫大鼠胃窦Cajal间质细胞自噬信号通路及胃动力的影响。方法:42只SD大鼠随机分为对照组(15只)和建模组(27只)。采用腹腔注射链脲佐菌素联合高脂高糖饮食不规则喂养8周建立糖尿病胃轻瘫模型,将建模成功的大鼠随机分为模型组和电针组,每组12只。电针组采用电针“足三里”“三阴交”“梁门”穴治疗;对照组与模型组不予以任何治疗。干预结束后,检测各组大鼠血糖水平、尿糖水平、胃排空率和小肠推进率;激光共聚焦检测Cajal间质细胞自噬流;透射电镜观察Cajal间质细胞自噬状态及超微结构;Western blot法检测Cajal间质细胞中微管相关蛋白1轻链3(LC3)、p62、磷酯酰肌醇3-激酶(PI3K)、三磷酸磷脂酰肌醇(PIP3)、蛋白激酶B(PKB/AKT)、腺苷酸活化蛋白激酶(AMPK)、哺乳动物雷帕霉素靶蛋白(mTOR)及磷酸化mTOR(p-mTOR)蛋白表达量。结果:与对照组比较,模型组大鼠胃排空率和小肠推进率显著下降(P<0.01),血糖和尿糖水平显著升高(P<0.01),AMPK表达下降(P<0.05),LC3-II/LC3-I、p62、PI3K、PIP3、AKT、mTOR和p-mTOR蛋白水平显著升高(P<0.05或P<0.01),绿色荧光蛋白(GFP)和红色荧光蛋白(RFP)的荧光强度均增强(P<0.01),Cajal间质细胞内可见1个自噬小体且胞内破损严重;与模型组比较,电针组大鼠胃排空率和小肠推进率显著上升(P<0.01),血糖和尿糖水平降低(P<0.05或P<0.01),AMPK表达上调(P<0.05),LC3-II/LC3-I、p62、PI3K、PIP3、AKT、mTOR和p-mTOR蛋白水平显著降低(P<0.05或P<0.01),GFP和RFP荧光强度均减弱(P<0.01),Cajal间质细胞内受损状况明显减轻。结论:电针能够提高糖尿病胃轻瘫大鼠胃肠动力,其作用机制可能与调节mTOR介导的信号通路而激活Cajal间质细胞自噬有关。AIM:This study aimed to observe the effects of electroacupuncture(EA)on the autophagy signaling pathway of interstitial cells of Cajal in the gastric antrum of diabetic gastroparesis(DGP)rats and to explore the possible mechanism.METHODS:A total of 42 Sprague-Dawley(SD)rats were randomly divided into normal control(CON)group(n=15)and model group(n=27).The rats in model group were intraperitoneally injected with 2%streptozotocin and fed with high-glucose and high-fat diet for 8 weeks to develop DGP rat model.Thereafter,the DGP rats were randomly divided into model group(DGP group)and EA group,with 12 rats in each group.The rats in EA group were treated with EA at"Zusanli"(ST 36),"Sanyinjiao"(SP 6),and"Liangmen"(ST 21),while the rats in CON and DGP groups were not provided any treatment.After the intervention was completed,the blood glucose level,urine glucose level,gastric emptying rate and small intestinal propulsion rate of the rats in each group were investigated.Autophagy flow was detected by immunofluorescence,and autophagy status and ultrastructure were detected by transmission electron microscopy.The protein levels of microtubule-associated protein 1 light chain 3(LC3),p62,phosphoinositol 3-kinase(PI3K),phosphatidylinositol-3,4,5-trisphosphate(PIP3),protein kinase B(PKB/AKT),mammalian rapamycin target protein(mTOR),phosphorylated mTOR(p-mTOR)and AMP-activated protein kinase(AMPK)were determined by Western blot.RE⁃SULTS:Compared with CON group,the blood and urine glucose levels were significantly increased,while the gastric emptying rate and small intestinal propulsion rate were significantly decreased in DGP group(P<0.01).The interstitial cells of Cajal were seriously damaged,one autophagosome was observed,and the autophagy flux in the interstitial cells of Cajal was impaired.The protein levels of LC3II/LC3-I,p62,PI3K,PIP3,AKT,mTOR and p-mTOR were significantly increased,while the protein expression level of AMPK was significantly decreased(P<0.01 or P<0.05).In comparison with DGP group,the blood and urin

关 键 词：电针 糖尿病胃轻瘫 CAJAL间质细胞 自噬 

分 类 号：R587.1[医药卫生—内分泌] R573[医药卫生—内科学]