摘要
目的:探讨烟酰胺腺嘌呤二核苷酸磷酸氧化酶2(NOX2)、核苷酸结合寡聚化结构域样受体家族吡啉结构域蛋白3(NLRP3)在肝纤维化患者血清中的表达及生物学意义。方法:选取2017年9月至2020年9月我院收治的80例肝纤维化患者为研究对象,同期选取80例我院收治的酒精性肝炎患者为对照组。采用酶联免疫吸附(ELISA)法检测受试者血清中NOX2、NLRP3的表达,并比较肝纤维化患者与对照组患者血清中NOX2、NLRP3的表达差异。体外培养肝星状细胞(HSC)系LX-2,用6 ng/ml转化生长因子-β1(TGF-β1)刺激24 h,活化并分为Control组、NC组、NOX2 siRNA组;实时荧光定量聚合酶链反应(qRT-PCR)检测细胞中NOX2、NLRP3的mRNA表达水平;细胞计数试剂盒(CCK-8)法检测细胞增殖抑制率;流式细胞术检测细胞凋亡率;2’,7’-二氯二氢荧光素二乙酸酯(DCFH-DA)法检测细胞ROS水平;蛋白免疫印迹(WB)检测增殖蛋白CyclinD1、CDK4,凋亡蛋白Caspase-3、Caspase-9及NOX2、NLRP3、IL-18、IL-1β的表达。结果:与对照组患者比较,肝纤维化组患者血清NOX2、NLRP3的表达显著升高(P<0.05);与Control组比较,NOX2 siRNA组细胞NOX2、NLRP3的mRNA表达、细胞内ROS含量,增殖蛋白CyclinD1、CDK4及蛋白NOX2、NLRP3、IL-18、IL-1β的表达均显著降低,细胞增殖抑制率、凋亡率、凋亡蛋白Caspase-3、Caspase-9的表达显著升高(P<0.05)。结论:NOX2、NLRP3可能参与肝纤维化的发生,其可能是治疗肝纤维化的靶点。
Objective:We aimed to investigate the expression and biological significance of nicotinamide adenine dinucleotide phosphate oxidase 2(NOX2) and nucleotide-binding oligomerization domain like receptor family pyrin domain containing 3(NLRP3) in liver fibrosis.Methods:Eighty patients with liver fibrosis in our hospital from September 2017 to September 2020 were selected as the research objects,and eighty patients with alcoholic hepatitis in our hospital were selected as the control group at the same time.The expression of NOX2 and NLRP3 in serum was detected by enzyme linked immunosorbent assay(ELISA),and the expression of NOX2 and NLRP3 in serum of patients with liver fibrosis and control group were compared.Hepatic stellate cell(HSC) line LX-2 was cultured in vitro and stimulated with 6 ng/mL transforming growth factor-β1(TGF-β1) for 24 hours to activate LX-2,and the cells were divided into control group,NC group,and NOX2 siRNA group.The mRNA expression levels of NOX2 and NLRP3 were detected by real-time fluorescence quantitative PCR(qRT-PCR).The inhibition rate of cell proliferation was detected by cell counting kit(CCK-8);the apoptosis rate was detected by flow cytometry,the level of ROS was detected by 2’,7’-dichlorodihydrofluorescein diacetate(DCFH-DA);and the expression of CyclinD1,CDK4,Caspase-3,Caspase-9,NOX2,NLRP3,IL-18 and IL-1β were detected by Western blot.Results:Compared with the control group,the expression of serum NOX2 and NLRP3 in the liver fibrosis group was significantly increased(P<0.05).Compared with the control group,the mRNA expression of NOX2 and NLRP3,the content of ROS in the cells,the expression of CyclinD1,CDK4,NOX2,NLRP3,IL-18 and IL-1β in NOX2 siR-NA group was significantly decreased,and the proliferation inhibition rate,apoptosis rate,the expression of Caspase-3 and Caspase-9 in NOX2 siRNA group was significantly increased( P < 0.05).Conclusion:NOX2 and NLRP3 may be involved in the occurrence of liver fibrosis,and may be the research targets of liver fibrosis treatment.
作者
谭莉霞
魏书堂
闫春晓
王航宇
杨丙信
徐梦阳
仝甲钊
TAN Li-xia;WEI Shu-tang;YAN Chun-xiao(Department of Gastroenterology,the First Affiliated Hospital of Henan University,Henan,Kaifeng,475000,China)
出处
《中西医结合肝病杂志》
CAS
2022年第3期221-224,共4页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
河南省医学科技攻关计划项目(No.LHGJ20190531)。
