摘要
目的研究白藜芦醇(Res)对人结肠上皮(NCM460)细胞炎症损伤的保护作用及其对Notch1信号通路的影响。方法体外培养NCM460细胞株,采用脂多糖(LPS,1 mg/L)干预细胞构建炎性损伤模型,低、高剂量Res(40、120μmol/L)及Notch1激动剂NSC 22423(2μmol/L)处理细胞24 h后,采用CCK8法检测NCM460细胞相对存活率,ELISA实验测定细胞培养液中炎性因子IL-6、IL-1β及TNF-α水平,Western blot实验分析i NOS、COX-2、Notch1、Hes-1及Math-1蛋白表达水平。结果与空白对照组相比,LPS组细胞相对存活率明显降低(P <0. 05),炎性因子IL-6、IL-1β及TNF-α水平显著升高(P <0. 05),i NOS、COX-2、Notch1、Hes-1蛋白表达量明显升高、Math-1蛋白表达显著降低(P <0. 05);与LPS诱导组相比,Res组细胞相对存活率显著升高(P <0. 05),炎性因子IL-6、IL-1β及TNF-α水平显著降低(P <0. 05),i NOS、COX-2、Notch1、Hes-1蛋白表达量明显降低、Math-1蛋白表达显著升高(P <0. 05);而Res和NSC共同干预NCM460细胞可以逆转上述指标的变化。结论 Res可通过Notch1信号通路调节人结肠上皮细胞的增殖和分化,对溃疡性结肠炎具有保护作用。
Objective To investigate the protective effects of resveratrol(Res)on inflammatory injury in human colon epithelial(NCM460)cells and the activation of Notch1 signaling pathway.Methods NCM460 cells were cultured in vitro,and LPS was used to produce the model of inflammatory injury.NCM460 cells were treated with low-,and high-dose Res(40 and 120μmol/l)and Notch1 agonist NSC 22423(2μmol/L)for 24 h,the cell relative survival rates were detected by CCK8 assay.The inflammatory cytokines(IL-6,IL-1βand TNF-α)were determined by ELISA assay.The expression levels of iNOS,COX-2,Notch1,Hes-1 and Math-1 were measured by Western blot.Results Compared with control group,the survival rates were significantly decreased(P<0.05),the levels of IL-6,IL-1βand TNF-αwere significantly increased(P<0.05),the expression levels of iNOS,COX-2,Notch1 and Hes-1 proteins were significantly increased,while the expression level of Math-1 was significantly decreased(P<0.05).Compared with LPS-stimulated group,the survival rates in Res and NSC groups were significantly increased(P<0.05),the levels of IL-6,IL-1βand TNF-αwere significantly decreased(P<0.05),the expression levels of iNOS,COX-2,Notch1 and Hes-1 proteins were significantly decreased,while the expression level of Math-1 was significantly increased(P<0.05).Moreover,NCM460 cells were treated with Res and NSC can reverse the LPS-induced changes.Conclusion Res can regulate the proliferation of human colon epithelial cells through Notch1 signaling pathway.
作者
赵丽
华夏
谭晔
胡承莲
熊刚
Zhao Li;Hua Xia;Tan Ye;Hu Chenglian;Xiong Gang(Department of Emergency,Central Hospital of En-shi Autonomous Prefecture,Enshi Hubei 445000,China)
出处
《遵义医学院学报》
2018年第6期695-699,共5页
Journal of Zunyi Medical University
基金
湖北省优秀青年科技创新团队计划项目(NO:T201705)
