脾胃培源方含药血清对鹅去氧胆酸诱导胃上皮细胞肠化生的影响  

Effect of decoction for reinforcing origin of spleen-stomach medicated serum on chenodeoxycholic acid-induced intestinal metaplasia in gastric epithelial cells

作　　者：裴蓓 魏思源 张艺 郝文哲 郑艳敏 李学军[2] PEI Bei;WEI Siyuan;ZHANG Yi;HAO Wenzhe;ZHENG Yanmin;LI Xuejun(Graduated School of Anhui University of Chinese Medicine,Heifei 230000,Anhui,China;The Second Affiliated Hospital of Anhui University of Chinese Medicine,Heifei 230000,Anhui,China;Heifei Institutes of Physical Science,Chinese Academy of Sciences,Heifei 230000,Anhui,China)

机构地区：[1]安徽中医药大学研究生院,安徽合肥230000 [2]安徽中医药大学第二附属医院,安徽合肥230000 [3]中国科学院合肥物质科学研究院,安徽合肥230000

出　　处：《现代中西医结合杂志》2023年第20期2822-2828,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：安徽省重大疑难疾病中西医协同攻关项目(0708-2021)。

摘　　要：目的探究脾胃培源方含药血清对鹅去氧胆酸诱导的人胃上皮细胞(GES-1)肠上皮化生的影响及其潜在作用机制。方法①采用CCK-8法检测不同浓度鹅去氧胆酸(0,20,50,100,150,200μmol/L)、脾胃培源方含药血清(0,2.5%,5%,10%,20%,30%,40%)对GES-1细胞活性的影响,确定后续实验鹅去氧胆酸和脾胃培源方含药血清浓度。②实验分为空白对照组、鹅去氧胆酸组、鹅去氧胆酸+5%脾胃培源方含药血清组、鹅去氧胆酸+10%脾胃培源方含药血清组、鹅去氧胆酸+20%脾胃培源方含药血清组,CCK-8法检测各组GES-1细胞活性。③实验分为空白对照组、20%脾胃培源方含药血清组、鹅去氧胆酸组、鹅去氧胆酸+20%脾胃培源方含药血清组,ELISA法检测炎症因子与氧化应激相关指标水平[白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)、丙二醛(MDA)、超氧化物歧化酶(SOD)]。④实验分为空白对照组、鹅去氧胆酸组、鹅去氧胆酸+5%脾胃培源方含药血清组、鹅去氧胆酸+10%脾胃培源方含药血清组、鹅去氧胆酸+20%脾胃培源方含药血清组,分别采用Western blot法和RT-PCR法检测肠上皮化生相关分子标志物(CDX1、CDX2、MUC2)蛋白及mRNA表达情况。结果选择100μmol/L的鹅去氧胆酸和5%,10%,20%脾胃培源方含药血清进行后续实验。各浓度脾胃培源方含药血清组GES-1细胞活性均明显低于鹅去氧胆酸组(P均<0.05),且各浓度脾胃培源方含药血清组GES-1细胞活性呈剂量依赖性降低。与空白对照组比较,鹅去氧胆酸组IL-6、IL-8、TNF-α、MDA水平均明显升高(P均<0.05),SOD水平明显降低(P<0.05);与鹅去氧胆酸组比较,鹅去氧胆酸+20%脾胃培源方含药血清组IL-6、IL-8、TNF-α、MDA水平均明显降低(P均<0.05),SOD水平明显升高(P<0.05)。鹅去氧胆酸组CDX1、CDX2、MUC2蛋白及mRNA相对表达量均明显高于空白对照组(P均<0.05);鹅去氧胆酸+10%脾胃培源方含药血清组和鹅Objective It is to investigate the effect of decoction for reinforcing origin of spleen-stomach medicated serum(DROSSMS)on chenodeoxycholic acid(CDCA)-induced intestinal metaplasia(IM)of gastric epithelial cells(GES-1)and its potential mechanism of action.Methods①The effects of different concentrations of CDCA(0,20,50,100,150,200μmol/L)and DROSSMS(0,2.5%,5%,10%,20%,30%,and 40%)on the activity of GES-1 cells were detected by CCK-8 assay to determine the concentrations of CDCA and DROSSMS in the subsequent experiments.②The experiment was divided into blank control group,CDCA group,CDCA+5%DROSSMS group,CDCA+10%DROSSMS group,CDCA+20%DROSSMS group,and the activities of GES-1 cells of each group were detected by CCK-8 method.③The experiment was divided into blank control group,20%DROSSMS group,CDCA group,and CDCA+20%DROSSMS group,the levels of inflammatory factors and oxidative stress related indexes[interleukin-6(IL-6),interleukin-8(IL-8),tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),and superoxide dismutase(SOD)]were detected by ELISA.④The experiment was divided into blank control group,CDCA group,and CDCA+5%DROSSMS group,CDCA+10%DROSSMS group,and CDCA+20%DROSSMS group,the expressions of intestinal epithelial chemotaxis-related molecular markers(CDX1,CDX2,and MUC2)proteins and mRNA were respectively detected by Western blot and RT-PCR methods.Results The subsequent experiments were carried out with 100μmol/L CDCA and 5%,10%,and 20%DROSSMS.The GES-1 cell activity of DROSSMS group of each concentration was significantly lower than that of the CDCA group(all P<0.05),and the GES-1 cell activity of DROSSMS group of each concentration was decreased in a dose-dependent manner.Compared with the blank control group,the levels of IL-6,IL-8,TNF-αand MDA in the CDCA group were significantly higher(all P<0.05),and the level of SOD was significantly lower(P<0.05);compared with the DROSSMS groups,the levels of IL-6,IL-8,TNF-α,and MDA in the CDCA+20%DROSSMS group were significantly lower(all P<0.05),and the level

关 键 词：肠上皮化生 脾胃培源方 鹅去氧胆酸 人胃上皮细胞 

分 类 号：R284[医药卫生—中药学]