摘要
目的探讨孟鲁司特预处理对阿司匹林诱导的胃黏膜上皮细胞GES-1损伤的影响及其作用机制。方法将GES-1细胞分为空白组、阿司匹林组、孟鲁司特低浓度组、孟鲁司特中浓度组、孟鲁司特高浓度组、孟鲁司特+阿司匹林组、孟鲁司特+阿司匹林+脂多糖组。阿司匹林组使用阿司匹林处理GES-1细胞;孟鲁司特低浓度组、孟鲁司特中浓度组、孟鲁司特高浓度组的GES-1细胞分别经10μmol/L、20μmol/L、40μmol/L孟鲁司特预处理后使用阿司匹林干预;孟鲁司特+阿司匹林组、孟鲁司特+阿司匹林+脂多糖组GES-1细胞经40μmol/L孟鲁司特预处理后,分别使用阿司匹林、阿司匹林+脂多糖干预。阿司匹林处理24 h后,检测各组细胞增殖抑制率、细胞周期、细胞凋亡率,细胞上清液乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和丙二醛水平,以及肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平;并检测细胞周期蛋白D1(CyclinD1)、细胞周期蛋白依赖性激酶2(CDK2)蛋白及mRNA表达量,以及B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved-Caspase-3)、细胞核转录因子κB抑制蛋白α(IκBα)、磷酸化IκBα(p-IκBα)、p65、磷酸化p65(p-p65)蛋白的表达量。结果(1)与空白组比较,阿司匹林组的细胞增殖抑制率、G_(0)/G_(1)期细胞比例、细胞凋亡率升高,细胞周期调控因子(CyclinD1和CDK2)mRNA和蛋白表达水平降低,促凋亡蛋白(Bax和Cleaved-Caspase-3)、核因子κB通路蛋白(p-IκBα和p-p65)的表达量以及LDH、丙二醛、炎症因子(TNF-α、IL-6)水平均升高,S期细胞比例、Bcl-2蛋白表达量和SOD活性降低(均P<0.05)。阿司匹林组、孟鲁司特低浓度组、孟鲁司特中浓度组、孟鲁司特高浓度组的细胞增殖抑制率、G_(0)/G_(1)期细胞比例、细胞凋亡率依次下降,细胞周期调控因子的mRNA和蛋白表达水平依次升高,促凋亡蛋白、核因子κB通路蛋白的表达量以及LDH、丙二醛、炎症因子水平依次降低,S期细胞比例、Bcl-2蛋白表达量和SOD活性依次升高(均P<0.05)。(2)与孟鲁司特+阿司匹林组比较,孟鲁司特+阿司匹林+脂多糖组的细胞增殖抑制率、细胞凋亡率及促凋亡相关因子表达均升高,炎症因子及核因子κB信号通路相关蛋白的表达均上调(均P<0.05),出现细胞周期阻滞。结论采用孟鲁司特预处理可减轻阿司匹林诱导的GES-1细胞损伤,其作用机制可能与减轻氧化损伤和炎症反应、调控核因子κB信号通路有关。
Objective To investigate the effect of preconditioning with montelukast on gastric mucosal epithelial cell injury induced by aspirin and its action mechanism.Methods GES-1 cells were divided into blank group,aspirin group,low-concentration montelukast group,medium-concentration montelukast group,high-concentration montelukast group,montelukast+aspirin group,montelukast+aspirin+lipopolysaccharide(LPS)group.GES-1 cells in the aspirin group were treated with aspirin;in the low-concentration montelukast group,medium-concentration montelukast group and high-concentration montelukast group,GES-1 cells were treated with aspirin after preconditioning with 10μmol/L,20μmol/L and 40μmol/L montelukast,respectively;GES-1 cells in the montelukast+aspirin group and montelukast+aspirin+LPS group received precondition with 40μmol/L montelukast,then were treated with aspirin and aspirin+LPS,respectively.After 24 hours of treatment with aspirin,the cell proliferation inhibition rate,cell cycle,cell apoptosis rate,the levels of lactate dehydrogenase(LDH),superoxide dismutase(SOD)and malondialdehyde in cell supernatants,as well as the levels of tumor necrosis factorα(TNF-α)and interleukin 6(IL-6),were detected in each group;besides,the following indicators were determined:the protein and mRNA expression of CyclinD1 and cyclin-dependent kinase 2(CDK2),and the expression of B-cell lymphoma 2(Bcl-2),Bcl-2 associated X protein(Bax),cleaved cysteinyl aspartate specific proteinase 3(Cleaved-Caspase-3),inhibitor of nuclear factorκBα(IκBα),phosphorylated IκBα(p-IκBα),p65 and phosphorylated p65(p-p65)proteins.Results(1)By comparison with the blank group,the aspirin group possessed increased cell proliferation inhibition rate,ratio of G_(0)/G_(1) phase cells and cell apoptosis rate,lower expression levels of cell cycle regulatory factors(CyclinD1 and CDK2)mRNAs and proteins,higher expression of apoptogenic proteins(Bax and cleaved-Caspase-3)and nuclear factorκB pathway related proteins(p-IκBαand p-p65),elevated levels of LDH,malondialdehyde and inflammatory factors(TNF-αand IL-6),lower proportion of S phase cells,decreased Bcl-2 protein expression and SOD activity(all P<0.05).In the aspirin group,low-,medium-and high-concentration montelukast groups,the cell proliferation inhibition rate,ratio of G_(0)/G_(1) phase cells and cell apoptosis rate decreased successively,and the expression levels of cell cycle regulatory factors mRNAs and proteins rose successively,the expression of apoptogenic proteins and nuclear factorκB pathway related proteins,as well as the levels of LDH,malondialdehyde and inflammatory factors declined successively,whereas the proportion of S phase cells,Bcl-2 protein expression and SOD activity increased successively(all P<0.05).(2)Compared with the montelukast+aspirin group,the montelukast+aspirin+LPS group yielded higher cell proliferation inhibition rate,cell apoptosis rate and expression of pro-apoptosis related factors,possessed up-regulated expression of inflammatory factors and nuclear factorκB signaling pathway-related proteins(all P<0.05),and exhibited cell cycle arrest.Conclusion Preconditioning with montelukast can attenuate GES-1 cell injury induced by aspirin,and the mechanism may be related to alleviation of oxidative damage and inflammation response,and regulation of nuclear factorκB signaling pathway.
作者
黄娇
罗杰伟
韩丽
杨莉
魏莹
HUANG Jiao;LUO Jie-wei;HAN Li;YANG Li;WEI Ying(Department of Pharmacy,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China;School of Basic Medicine,North Sichuan Medical College,Nanchong 637800,China;School of Pharmacy,North Sichuan Medical College,Nanchong 637800,China)
出处
《广西医学》
CAS
2021年第11期1352-1359,共8页
Guangxi Medical Journal
基金
四川省南充市市校战略合作科技项目(18SXHZ0133)。
关键词
胃黏膜损伤
阿司匹林
孟鲁司特
胃黏膜上皮细胞
核因子ΚB信号通路
细胞增殖
氧化应激
炎症
Gastric mucosal injury
Aspirin
Montelukast
Gastric mucosal epithelial cell
Nuclear factorκB signaling pathway
Cell proliferation
Oxidative stress
Inflammation
作者简介
黄娇(1990~),女,本科,药师,研究方向:临床药学。电子邮箱:jxkcg73@163.com;通信作者:魏莹(1987~),女,硕士,讲师,研究方向:药物分析,电子邮箱:sjxjj33@163.com。
