柴胡皂苷-d对H_(2)O_(2)诱导的HL-7702细胞ROS产生及NLRP3炎症体的影响  

Effect of saikosaponin-d on ROS production and NLRP3 inflammatory body in HL-7702 cells induced by H_(2)O_(2)

作　　者：周蒙恩 张娜[1] 阙任烨 林柳兵[1] 李勇[1] ZHOU Meng-En;ZHANG Na;QUE Ren-Ye;LIN Liu-Bing;LI Yong(Department of Gastroenterology,Shanghai Municipal Hospital of Traditional Chinese Medicine Affiliated to Shanghai University of Traditional Chinese Medi-cine,Shanghai 200071,China)

机构地区：[1]上海中医药大学附属市中医医院脾胃病科,上海200071

出　　处：《中国免疫学杂志》2021年第20期2470-2474,2481,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(81873157,81573775,81573775)。

摘　　要：目的:探讨柴胡皂-d(SSd)对H_(2)O_(2)诱导HL-7702细胞ROS产生及NLRP3炎症体表达的影响。方法:体外培养HL-7702细胞,采用H_(2)O_(2)诱导HL-7702细胞造模,并分为空白对照组,H_(2)O_(2)模型组,MitoTEMPOL+H_(2)O_(2)组,SSd+H_(2)O_(2)组,使用荧光检测线粒体膜电位(JC-1)、MitoSox Red线粒体超氧化物阴离子表达,化学发光法检测ATP水平,Western blot、RT-PCR法检测NLRP3炎症体的表达。结果:与对照组比较,H_(2)O_(2)对HL-7702细胞的氧化应激损伤使线粒体膜电位有下降趋势(P<0.05),ATP生成减少(P<0.05),MitoSOX Red荧光强度增强(P<0.05),NLRP3、ASC、Caspase1蛋白及基因表达明显上调(P<0.05);与模型组比较,使用MitoTEMPOL、SSd干预后线粒体膜电位明显上升(P<0.05),ATP明显上升(P<0.05),MitoSOX Red荧光强度下降(P<0.05),NLRP3、ASC、Caspase1蛋白及基因表达明显下调(P<0.05)。结论:H_(2)O_(2)诱导的HL-7702细胞中线粒体来源的ROS可以诱导NLRP3炎症体的活化,SSd可以通过抑制H_(2)O_(2)诱导的HL-7702细胞ROS的产生及NLRP3炎症体的表达,发挥保护肝细胞作用。Objective:To explore the effect of saikosaponin-d(SSd)on the production of ROS and the expression of NLRP3 in-flammatory body in HL-7702 cells induced by H_(2)O_(2).Methods:HL-7702 cells were cultured in vitro,and HL-7702 cells were induced by H_(2)O_(2).HL-7702 cells were divided into blank control group,H_(2)O_(2)model group,MitoTEMPOL+H_(2)O_(2)group,SSd+H_(2)O_(2)group.The mitochondrial membrane potential(JC-1)and the expression of superoxide anion in MitoSox Red mitochondria were detected by fluo-rescence,the level of ATP was detected by chemiluminescence,and the expression of NLRP3 inflammatory body was detected by Western blot and RT-PCR.Results:Compared with the control group,the mitochondrial membrane potential of HL-7702 cells induced by H_(2)O_(2)decreased after oxidative stress(P<0.05).The production of ATP decreased(P<0.05).The fluorescence intensity of MitoSOX Red increased(P<0.05).The protein and gene expression of NLRP3,ASC and Caspase1 were significantly up-regulated(P<0.05).Compared with the model group,the mitochondrial membrane potential increased significantly after the intervention of MitoTEMPOL and SSd(P<0.05).ATP increased significantly(P<0.05).The fluorescence intensity of MitoSOX Red decreased significantly,and the protein and gene expression of NLRP3,ASC,Caspase1 decreased significantly(P<0.05).Conclusion:Mitochondrial ROS in HL7702 cells induced by H_(2)O_(2)can induce the activation of NLRP3 inflammatory body.SSd can protect hepatocytes by inhibiting the pro-duction of ROS and the expression of NLRP3 inflammatory bodies in HL-7702 cells induced by H_(2)O_(2).

关 键 词：柴胡皂-d HL-7702细胞 ROS NLRP3炎症体 

分 类 号：R284[医药卫生—中药学]