五味子乙素预处理对过氧化氢诱导的H9c2细胞焦亡及TXNIP/NLRP3/Caspase-1通路的影响  

Effect of Schisandrin B pretreatment on H 2O 2-induced pyroptosis and TXNIP/NLRP3 pathway in H9C2 cells

作　　者：王瑞婷[1] 赵松[2] 郝娜[3] 李立萍[3] 解丽君 WANG Ruiting;ZHAO Song;HAO Na;LI Liping;XIE Lijun(Institute of Chinese Herbs of Chengde Medical University,Chengde 071000,Hebei,China;College of Basic Medical Sciences,Hebei Medical University,Shijiazhuang 050017,Hebei,China;Institute of MateriaMedica of Hebei Provincial Center for Disease Control and Prevention,Shijiazhuang 050021,Hebei,China)

机构地区：[1]承德医学院中药研究所,河北承德071000 [2]河北医科大学基础医学院,河北石家庄050017 [3]河北省疾病预防控制中心药物研究所,河北石家庄050021

出　　处：《现代中西医结合杂志》2024年第13期1807-1812,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：河北省科技厅应用基础研究重点项目(179676127D)。

摘　　要：目的基于硫氧还蛋白互作蛋白(TXNIP)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)/半胱氨酸蛋白酶-1(Caspase-1)通路观察五味子乙素对过氧化氢(H 2O 2)诱导的大鼠H9c2心肌细胞氧化损伤及焦亡的影响,探讨五味子乙素的心肌保护作用机制。方法体外培养H9c2心肌细胞,实验设4组:正常组细胞常规孵育,五味子乙素组细胞加入40μmol/L五味子乙素孵育24 h,H 2O 2组细胞加入1000μmol/L的H 2O 2孵育30 min,H 2O 2+五味子乙素组细胞加入40μmol/L五味子乙素孵育24 h后再加入H 2O 2孵育30 min。CCK-8法检测细胞活力,DCFH-DA探针检测细胞中活性氧(ROS)含量,试剂盒检测细胞培养上清中乳酸脱氢酶(LDH)含量和细胞中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)含量,Western blot法检测细胞中TXNIP、硫氧还蛋白(Trx)、NLRP3、裂解的半胱氨酸蛋白酶-1(Cleaved Caspase-1)、消皮素D(GSDMD)-N蛋白表达情况,ELISA法检测细胞培养上清中白细胞介素-18(IL-18)、白细胞介素-1β(IL-1β)含量。结果与正常组比较,H 2O 2组细胞活力明显下降(P<0.05),细胞中ROS、MDA含量和TXNIP、NLRP3、Cleaved Caspase-1、GSDMD-N蛋白相对表达量及细胞培养上清中LDH、IL-1β、IL-18含量均明显升高(P均<0.05),细胞中SOD、GSH-Px含量和Trx蛋白相对表达量均明显降低(P均<0.05);与H 2O 2组比较,H 2O 2+五味子乙素组的细胞活力明显升高(P<0.05),细胞中ROS、MDA含量和TXNIP、NLRP3、Cleaved Caspase-1、GSDMD-N蛋白相对表达量及细胞培养上清中LDH、IL-1β、IL-18含量均明显降低(P均<0.05),细胞中SOD、GSH-Px含量和Trx蛋白相对表达量均明显升高(P均<0.05)。结论五味子乙素可能通过影响TXNIP/NLRP3/Caspase-1通路,从而减轻H 2O 2诱导的H9c2细胞氧化损伤,抑制心肌细胞焦亡。Objective It is to observe the effects of Schisandrin B on the oxidative damage and pyroptosis of H 2O 2-induced H9c2 cardiomyocytes in rats via thioredoxin-interacting protein(TXNIP)/NOD-like receptor protein 3(NLRP3)/Caspase-1 pathway,and to explore its mechanism of myocardial protective effects.Methods H9c2 cardiomyocytes were cultured in vitro,and were divided into 4 groups for the experiment:the cells in the normal group were routinely incubated,the cells in the Schisandrin B group were incubated for 24 h with 40μmol/L Schisandrin B,the cells in the H 2O 2 group were incubated for 30 min with 1000μmol/L H 2O 2,and the cells in the H 2O 2+Schisandrin B group were firstly incubated with 40μmol/L Schisandrin B for 24 h and then with H 2O 2 for another 30 min.the cell viability was detected by CCK-8 assay,the content of reactive oxygen species(ROS)was detected by DCFH-DA probe,the contents of lactate dehydrogenase(LDH)in cell culture supernatant,and the contents of malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)in the cells were detected by kits,the protein expressions of TXNIP,thioredoxin(Trx),NLRP3,cleaved Caspase-1,glucocorticoid D(GSDMD-N)in the cells were detected by Western blot,and the levels of interleukin-18(IL-18)and interleukin-1β(IL-1β)in cell culture supernatant were detected by ELISA.Results Compared with the normal group,the cell viability was significantly decreased in the H 2O 2 group(P<0.05),and the contents of ROS,MDA and the relative protein expressions of TXNIP,NLRP3,cleaved Caspase-1,GSDMD-N in the cells and the contents of LDH,IL-1βand IL-18 in the cell culture supernatant were significantly increased(all P<0.05),and the contents of SOD,GSH-Px content and relative protein expression of Trx in the cells were significantly decreased(all P<0.05).Compared with the H 2O 2 group,the cell viability was significantly increased in the H 2O 2+Schisandrin B group(P<0.05),and the contents of ROS,MDA and the relative protein expressions of TXNIP,NLRP3,cleaved Caspase

关 键 词：五味子乙素 H9C2 氧化应激 焦亡 硫氧还蛋白互作蛋白 核苷酸结合寡聚化结构域样受体蛋白3 

分 类 号：R-332[医药卫生]