半夏泻心汤调节胃电节律失常大鼠胃窦Cajal间质细胞自噬的作用  

Banxia Xiexintang Regulates Autophagy of Interstitial Cells of Cajal in Gastric Antrum of Rats with Gastric Electric Dysrhythmia

作　　者：张迪 李雨静 吉静 张冬梅[2] 陈萌 ZHANG Di;LI Yujing;JI Jing;ZHANG Dongmei;CHEN Meng(School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学中医学院,北京100029 [2]北京中医药大学东直门医院,北京100700

出　　处：《中国实验方剂学杂志》2023年第6期55-62,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82174270)。

摘　　要：目的:通过观察半夏泻心汤对于胃电节律失常大鼠胃窦组织中Cajal间质细胞(ICCs)自噬的影响,探究其保护作用及调节机制。方法:32只SD大鼠随机分为正常组、模型组、半夏泻心汤组(24.68 g·kg^(-1))及多潘立酮组(2.7 mg·kg^(-1))。通过“隔日进食+饮稀盐酸”法建立胃电节律失常大鼠模型,给予半夏泻心汤及阳性药多潘立酮进行干预。每周记录大鼠体质量;生物机能实验系统记录大鼠胃电活动;透射电子显微镜观察大鼠胃窦组织超微结构变化;免疫荧光双标法检测大鼠胃窦组织中酪氨酸激酶受体(c-kit)/自噬关键分子酵母Atg6同系物(Beclin1)共表达情况;蛋白免疫印迹法(Western blot)检测胃窦组织中微管相关蛋白1轻链3(LC3)B、p62蛋白表达,并计算LC3BⅡ/Ⅰ。结果:与正常组比较,模型组大鼠体质量减轻(P<0.01);胃电主频率、主功率下降(P<0.01);电镜下观察到胃窦ICCs超微结构损伤,细胞器变性坏死,可见少量自噬泡;免疫荧光双标结果显示,胃窦组织中c-kit阳性表达减低,Beclin1阳性表达增加;Western blot结果显示,胃窦组织中LC3BⅡ/Ⅰ增加(P<0.01),p62蛋白表达减低(P<0.01)。与模型组比较,半夏泻心汤及多潘立酮能够增加大鼠体质量(P<0.01),升高胃电主频率、主功率(P<0.01),修复胃窦ICCs超微结构损伤,上调胃窦组织中c-kit阳性表达并下调Beclin1阳性表达,上调胃窦组织中p62表达(P<0.05);半夏泻心汤能够抑制胃窦组织中LC3BⅠ转化为LC3BⅡ(P<0.05)。结论:半夏泻心汤能够通过调节自噬相关蛋白的表达,降低ICCs细胞自噬,调节ICCs的数量及结构,从而改善大鼠胃电节律失常,阐释了半夏泻心汤治疗“心下痞”的作用内涵。Objective:To observe the effect of Banxia Xiexintang on the autophagy of interstitial cells of Cajal(ICCs)in the gastric antrum of rats with gastric electric dysrhythmia,and explore the protective effect and regulatory mechanism.Method:Thirty-two SD rats were randomly assigned into a normal group,a model group,a Banxia Xiexintang(24.68 g·kg^(-1))group,and a positive drug(2.7 mg·kg^(-1))group.The rat model of gastric electric dysrhythmia was established by the method of dieting every other day and drinking dilute hydrochloric acid,and Banxia Xiexintang and the positive drug were administrated for intervention.The body weight of each rat was recorded weekly.The gastric electric activity was recorded by the biological function experimental system.The ultrastructural changes of the gastric antrum tissue were observed by a transmission electron microscope.The co-expression of receptor tyrosine kinase(c-kit)/mammalian homolog of yeast Atg6(Beclin1)in the gastric antrum tissue was detected by double immunofluorescence labeling method.The expression of microtubule-associated protein 1 light chain 3B(LC3B)and p62 protein in the gastric antrum tissue was determined by Western blot,and the LC3BⅡ/Ⅰratio was calculated.Result:Compared with the normal group,the modeling reduced the body weight(P<0.01)and decreased the dominant frequency and dominant power of gastric electricity(P<0.01).In addition,the modeling caused ultrastructural damage of ICCs in gastric antrum,degeneration and necrosis of organelles,and appearance of a small number of autophagic vesicles.The results of double immunofluorescence labeling showed that the modeling inhibited the positive expression of c-kit and promoted the positive expression of Beclin1 in gastric antrum tissue.Western blot results showed that the modeling increased the ratio of LC3BⅡ/Ⅰ(P<0.01)and down-regulated the expression of p62protein(P<0.01)in the gastric antrum tissue.Compared with the model group,Banxia Xiexintang and the positive drug increased the body weight(P<0.01)and

关 键 词：半夏泻心汤 CAJAL间质细胞 自噬 胃电节律失常 心下痞 

分 类 号：R2-0[医药卫生—中医学] R33R289R573