芪石升降归元颗粒对胃食管反流病大鼠的干预作用及可能作用机制  

Mechanism of Qishi Shengjiang Guiyuan Granule(芪石升降归元颗粒)on GERD Rats Based on Bcl-2/CytC/Caspase3 Pathway Mediating Esophageal Cell Apoptosis

作　　者：黎丽群 黄美祯 王金鑫 谭金晶[1] 刘园园[1] 谢胜[1] LI Liqun;HUANG Meizhen;WANG Jinxin;TAN Jinjing;LIU Yuanyuan;XIE Sheng(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530001,Guangxi,China;Graduate School of Guangxi University of Chinese Medicine,Nanning 530001,Guangxi,China)

机构地区：[1]广西中医药大学第一附属医院,广西南宁530001 [2]广西中医药大学研究生学院,广西南宁530001

出　　处：《辽宁中医杂志》2024年第12期178-183,I0018,共7页Liaoning Journal of Traditional Chinese Medicine

基　　金：广西科技厅自然科学基金青年基金项目(2021GXNSFBA196047,2021GXNSFBA196055);国家自然科学基金青年基金项目(82004299);广西中医药大学校级课题(2020QN011,2021QN018);国家中医临床研究基地建设项目(国中医药科技函20181131);广西中医药重点研究室建设项目脾胃治未病重点研究室(桂中医药科教发1202319号);国家中医药传承创新中心项目(发改办社会20221366号)。

摘　　要：目的分析芪石升降归元颗粒对胃食管反流病(gastroesophageal reflax disease,GERD)大鼠的干预作用,并基于B细胞淋巴瘤2(B cell lymphoma 2,Bcl-2)/细胞色素C(cytochrome C,Cyt C)/Caspase3通路探讨其作用机制。方法将42只SPF雄性SD大鼠随机分为假手术组、模型组、西药组、中药组。除假手术组10只外,其余各组均采用食管支架植入术制作GERD大鼠模型。造模成功后,西药组予以枸橼酸莫沙必利分散片联合兰索拉唑肠溶片灌胃,中药组给予芪石升降归元颗粒灌胃,疗程均为14 d。干预结束后,采用苏木素-伊红(HE)染色观察大鼠食管下段组织形态学变化;酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)法检测血清中Bcl-2、Caspase3及Cyt C的含量;实时荧光定量聚合酶链式反应(real-time fluorescence quantitative polymerase chain reaction,real-time PCR)检测食管组织内Bcl-2 mRNA、Caspase3 mRNA、Cyt C mRNA表达水平;蛋白免疫印迹法(Western blot)检测食管组织内Bcl-2、Caspase3、Cyt C蛋白表达水平。结果GERD模型大鼠精神状态差,活动及进食量减少,光镜下观察可见细胞排列紊乱,鳞状上皮增生,固有层乳头延伸,上皮细胞炎性浸润。经西药及芪石升降归元颗粒治疗后,大鼠精神状态佳,进食量及活动正常,光镜下观察可见细胞排列规整,鳞状上皮增生好转,无固有层乳头延伸,未见明显炎症细胞浸润。与假手术组比较,模型组食管组织中的Cyt C mRNA、Caspase3 mRNA及蛋白表达水平均升高(P<0.05,P<0.01,P<0.001);模型组血清Bcl-2表达量降低(P<0.001),Cyt C和Caspase3表达量升高(P<0.001,P<0.01)。与模型组比较,西药组食管组织中的Bcl-2 mRNA及蛋白表达水平升高(P<0.01,P<0.05),Caspase3 mRNA表达量降低(P<0.05);血清中Cyt C的浓度降低(P<0.001)。与模型组比较,中药组食管组织Bcl-2 mRNA及蛋白表达水平升高(P<0.05,P<0.001),Cyt C mRNA和Caspase3 mRNA及蛋白表达水平均降低(P<0.001);血清Bcl-Objective To explore the preventive and therapeutic effect and mechanism of Qishi Shengjiang Guiyuan Granule(芪石升降归元颗粒)on esophageal cell apoptosis in rats with gastroesophageal reflux disease(GERD)based on B cell lymphoma 2(Bcl-2)/cytochrome C(Cyt C)/Caspase3.Methods Forty-two SPF male SD rats were randomly divided into sham operation group,model group,western medicine group and Chinese medicine group.Except for sham operation group(n=10),esophageal stent implantation was used to make GERD rat model in other groups.After the successful establishment of the model,the western medicine group was given Mosapride Citrate Dispersible Tablets combined with Lansoprazole Enteric-Coated Tablets,and the Chinese medicine group was given Qishi Shengjiang Guiyuan Granule for 14 days.After intervention,the morphological changes of lower esophagus were observed by hematoxylin-eosin(HE)staining,the contents of Bcl-2,Caspase3 and Cyt C in serum were detected by enzyme-linked immunosorbent assay(ELISA),the expressions of Bcl-2 mRNA,Caspase3 mRNA and Cyt C mRNA in esophageal tissue were detected by real-time fluorescence quantitative polymerase chain reaction(Real-timePCR),and the protein expressions of Bcl-2,Caspase3 and Cyt C in esophageal tissue was detected by Western blot method.Results GERD model rats had poor mental state,decreased activity and food intake,cell arrangement disorder,squamous epithelial hyperplasia,lamina propria papilla extension,and inflammatory infiltration of epithelial cells were observed under light microscope.After treatment with western medicine and Qishi Shengjiang Guiyuan Granule,the mental state of rats was good,food intake and activity were normal,cell arrangement was orderly,squamous epithelial hyperplasia was improved,lamina propria papilla was not extended,and no obvious inflammatory cell infiltration was observed under light microscope.Compared with those of the sham operation group,mRNA and protein expression levels of Cyt and Caspase3 in esophageal tissues of the model group we

关 键 词：胃食管反流病 芪石升降归元颗粒 Bcl-2/Cyt C/Caspase3通路 凋亡 

分 类 号：R285[医药卫生—中药学]