摘要
目的 探讨原阿片碱对脂多糖(LPS)诱导的肠上皮细胞炎症、凋亡和氧化应激的影响。方法 通过四甲基偶氮唑蓝(MTT)实验筛选出5μg/ml的LPS处理NCM460细胞,同时加入不同浓度的原阿片碱(0、5、10、20μM)刺激细胞24 h。通过MTT法和流式细胞术检测NCM460细胞活力和凋亡率。通过Western blot免疫印迹法检测各处理组中凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)表达水平。通过ELISA检测炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的表达水平。通过ELISA检测抗氧化酶超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化能力(T-AOC)和髓过氧化物酶(MPO)的表达水平以及通过流式细胞术检测活性氧(ROS)表达水平,明确原阿片碱对NCM460细胞氧化应激反应的影响。结果 5μg/ml的LPS组的NCM460细胞活力低于阴性对照组,差异有统计学意义(P<0.05)。LPS+原阿片碱组NCM460细胞活力、Bcl-2蛋白高于LPS组,且呈剂量依赖性,差异有统计学意义(P<0.05)。LPS+原阿片碱组NCM460细胞的凋亡率、Bax蛋白、TNF-α、IL-1β和IL-6、氧化应激分子SOD、MDA、T-AOC、MPO和ROS的表达水平低于LPS组,且呈剂量依赖性,差异有统计学意义(P<0.05)。结论 原阿片碱可通过降低细胞凋亡、炎症反应及氧化应激反应改善LPS诱导的肠上皮细胞炎性损伤。
Objective To investigate the effects of protoopioid on inflammation,apoptosis and oxidative stress of intestinal epithelial cells induced by lipopolysaccharides(LPS).Methods NCM460 cells were screened by methyl thiazolyl tetrazolium(MTT) assay and treated with 5 μg/ml LPS,and the cells were stimulated with different concentrations of protoopioid(0,5,10,20 μM) for 24 h.The viability and apoptosis rate of NCM460 cells were detected by MTT assay and flow cytometry.The expression levels of apoptosis-associated protein B-celllymphoma-2(Bcl-2) and Bcl-2-associated X protein(Bax) in each treatment group were detected by Western blot.The expression levels of inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β) and interleukin-6(IL-6) were detected by ELISA.The expression levels of superoxidedismutase(SOD),malondialdehyde(MDA),total antioxidant capacity(T-AOC) and myeloperoxidase(MPO) were detected by ELISA and the expression levels of reactive oxygen species(ROS) were detected by flow cytometry to determine the effect of protoopioid on the oxidative stress response of NCM460 cells.Results The viability of NCM460 cells in the 5 μg/ml LPS group was lower than that in the negative control group,the difference was statistically significant(P<0.05).The cell viability and Bcl-2 protein of NCM460 in LPS+ protoopioid group were higher than those in LPS group in a dose-dependent manner,with statistical significances(P<0.05).The expression levels of apoptosis rate,Bax protein,TNF-α,IL-1β and IL-6,oxidative stress molecules SOD,MDA,T-AOC,MPO and ROS in NCM460 cells in LPS+ protoopioid group were lower than those in LPS group in a dose-dependent manner,with statistical significances(P<0.05).Conclusion Protoopioid can ameliorate LPS-induced inflammatory injury of intestinal epithelial cells by decreasing apoptosis,inflammatory response and oxidative stress response.
作者
徐忠军
李君玉
欧阳灿晖
谢云
叶艳清
XU Zhongjun;LI Junyu;OUYANG Canhui;XIE Yun;YE Yanqing(Department of Medical Imaging,the First Affiliated Hospital of Gannan Medical University,Jiangxi Province,Ganzhou 341000,China;Department of Gastroenterology,the First Affiliated Hospital of Gannan Medical University,Jiangxi Province,Ganzhou 341000,China)
出处
《中国当代医药》
2023年第36期5-10,共6页
China Modern Medicine
基金
江西省卫生健康委科技计划项目(202310741)。
关键词
原阿片碱
脂多糖
肠上皮细胞
凋亡
氧化应激
炎症反应
Protoopioid
Lipopolysaccharide
Intestinal epithelial cells
Apoptosis
Oxidative stress
Inflammatory response
