南蛇藤提取物对小鼠胃类器官生长及E-caherin表达的影响  

Effects of Celastrus orbiculatus extract on gastric organoids and expression of E-caherin in mice

作　　者：朱方圆 张效泽 刘延庆[2] 李平[1] 朱耀东 Zhu Fangyuan;Zhang Xiaoze;Liu Yanqing;Li Ping;Zhu Yaodong(Dept of Chinese Integrative Medicine Oncology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;College of Medicine,Yangzhou University,Yangzhou 225003)

机构地区：[1]安徽医科大学第一附属医院中西医结合肿瘤科,合肥230022 [2]扬州大学医学院,扬州225003

出　　处：《安徽医科大学学报》2023年第3期471-476,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82274355);安徽省自然科学基金(编号:2208085MH278);安徽高校自然科学研究项目(编号:KJ2020A0215)。

摘　　要：目的探讨南蛇藤提取物(COE)对小鼠胃类器官生长及其胃上皮细胞E-cadherin表达的影响。方法将8周龄C57小鼠胃幽门部分离并培养成胃类器官。光镜下观察胃类器官形成的动态变化,HE染色观察胃上皮细胞间结构,免疫荧光染色观察胃上皮细胞的上皮钙黏蛋白E-cadherin表达。类器官传代到第3代后,用不同浓度的COE(0、5、10、20μg/ml)处理,收集类器官,计算其数量,测量其直径,噻唑蓝(MTT)比色法测其细胞活性,并用Western blot法检测COE处理后类器官中的E-cadherin表达。结果24~48 h形成囊样结构并出现中心结构为囊腺样的三维细胞团,72 h后逐渐出芽并形成胃类器官,提示类器官成功构建。类器官中的上皮细胞标志物E-cadherin表达,进一步证实类器官的形成。与对照组比较,COE组胃类器官数量与直径均增加、细胞活性增强(P<0.05),且E-cadherin的表达随着COE的剂量增加而逐渐增加(P<0.01)。结论小剂量的COE可促进E-cadherin的表达与类器官的生长形成,可能会影响胃黏膜损伤的修复。Objective To investigate the effect of Celastrus orbiculatus extract(COE)on the growth of gastric organoids and the expression of E-cadherin in gastric epithelial cells of mice.Methods The gastric pylorus of 8-week-old C57 mice was isolated and cultured into gastric organoids.The dynamic changes of gastric organoid formation were observed under light microscope;the intercellular structure of gastric epithelium was observed by HE staining;the expression of epithelial-cadherin E-cadherin in gastric epithelial cells was observed by immunofluorescence staining.After passage to the third-generation,the organoids were treated with different concentrations of COE(0,5,10,20μg/ml),the organoids were collected,their numbers were counted,their diameters were measured,their cellular activities were measured by methyl thiazolyl tetrazolium(MTT)colorimetry,and Western blot was used to detect the expression of E-cadherin in organoids after COE treatment.Results At 24 to 48 h,cystlike structures were formed and three-dimensional cell clusters with cystic gland-like central structure appeared,and gradually budding and forming gastric organoids after 72 h,suggesting that the organoids were successfully constructed.The epithelial cell marker E-cadherin was expressed in the organoid,which further confirmed the formation of organoid.Compared with the control group,the number and diameter of gastric organoids in the COE group significantly increased,cell activity was significantly enhanced(P<0.05),and the expression of E-cadherin increased with the increase of COE dose(P<0.01).Conclution Low dose COE can promote the expression of E-cadherin and the growth and formation of organoids,which may affect the repair of gastric mucosa injury.

关 键 词：南蛇藤 胃类器官 小鼠 E-CADHERIN 

分 类 号：R285[医药卫生—中药学]