摘要
目的探究缬沙坦(VAL)调节活性氧(ROS)/核苷酸结合寡聚结构域样受体蛋白3(NLRP3)/胱天蛋白酶1(caspase-1)信号通路对缺氧复氧(H/R)诱导心肌细胞焦亡的影响。方法用浓度为0.1~3.2μmol/L的缬沙坦处理后,H/R处理H9C2细胞,MTT法测细胞活性,筛选缬沙坦最佳浓度;将H9C2细胞分为Control组,H/R组,缬沙坦低、中、高浓度组(VAL-L组、VAL-M组、VAL-H组),缬沙坦高浓度+ROS激活剂组(VAL-H+TMAO组);用JC-1染色法检测细胞线粒体膜电位(MMP)水平;用ELISA法检测H9C2细胞丙二醛(MDA)含量、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)、活性氧(ROS)含量;Western blot检测IL-1β、IL-18、NLRP3、caspase-1蛋白表达。结果浓度为0.1~3.2μmol/L的缬沙坦可促进H/R诱导的H9C2细胞增殖,选择缬沙坦浓度为0.4μmol/L、0.8μmol/L、1.6μmol/L进行实验。与Control组相比,H/R组ROS、LDH、MDA、PI阳性率、IL-1β、IL-18、ROS、NLRP3、caspase-1表达水平明显升高,MMP、SOD水平降低(P<0.001);与H/R组比较,VAL-L、VAL-M、VAL-H组H9C2细胞ROS、LDH、MDA、PI阳性率、IL-1β、IL-18、ROS、NLRP3、caspase-1表达水平降低,MMP、SOD水平上升(P<0.001);与VAL-H组比较,VAL-H+TMAO组H9C2细胞ROS、LDH、MDA、PI阳性率、IL-1β、IL-18、ROS、NLRP3、caspase-1表达水平明显上升,MMP、SOD水平下降(P<0.001)。结论缬沙坦可能抑制ROS/NLRP3/caspase-1信号通路抑制H/R诱导心肌细胞焦亡。
Objective To investigate the effect of valsartan(VAL)on hypoxia/reoxygenation(H/R)induced cardiomyocyte pyroptosis by regulating the reactive oxygen species(ROS)/nucleotide binding oligomeric domain like receptor protein 3(NLRP3)/caspase-1 signaling pathway.Methods After being treated with 0.1~3.2μmol/L valsartan,H9C2 cells were treated with H/R,cell activity was measured by MTT assay,and the optimal concentration of valsartan was selected.H9C2 cells were divided into control group,H/R group,valsartan low,medium and high concentration groups(VAL-L group,VAL-M group,VAL-H group),and valsartan high concentration+ROS activator group(VAL-H+TMAO group).JC-1 staining method was applied to detect the level of mitochondrial membrane potential(MMP).ELISA method was applied to detect the contents of malondialdehyde(MDA),superoxide dismutase(SOD),lactate dehydrogenase(LDH)and reactive oxygen species(ROS)in H9C2 cells.Western blot was applied to detect the expression of IL-1β,IL-18,NLRP3,caspase-1 protein.Results Valsartan at concentrations of 0.1-3.2μmol/L was able to promote H/R induced proliferation of H9C2 cells.Valsartan concentrations of 0.4μmol/L,0.8μmol/L,and 1.6μmol/L were selected for the experiment.Compared with Control group,the positive rates of ROS,LDH,MDA and PI,the expression levels of IL-1β,IL-18,ROS,NLRP3 and caspase-1 in H/R group were significantly increased,and the levels of MMP and SOD were decreased(P<0.001).Compared with H/R group,the positive rates of ROS,LDH,MDA and PI,the expression levels of IL-1β,IL-18,ROS,NLRP3 and caspase-1 in H9C2 cells in VAL-L,VAL-M and VAL-H groups were decreased,while the levels of MMP and SOD were increased(P<0.001).Compared with VAL-H group,the positive rates of ROS,LDH,MDA,PI,IL-1β,IL-18,ROS,NLRP3 and caspase-1 in H9C2 cells in VAL-H+TMAO group were significantly increased,while the levels of MMP and SOD were decreased(P<0.001).Conclusion Valsartan may inhibit ROS/NLRP3/caspase-1 signaling pathway and inhibit H/R-induced cardiomyocyte pyroptosis.
作者
陈晨
陈莉洁
CHEN Chen;CHEN Lijie(Department of the First Outpatient,the 960 Hospital of the Joint Logistic Support Force,Jinan 250000,Shandong,China)
出处
《医学研究与战创伤救治》
CAS
北大核心
2024年第9期909-913,共5页
Journal of Medical Research & Combat Trauma Care
关键词
心肌细胞
缬沙坦
活性氧/核苷酸结合寡聚结构域样受体蛋白3/胱天蛋白酶1
缺氧复氧
焦亡
cardiomyocytes
valsartan
reactive oxygen species(ROS)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)/caspase-1
hypoxia/reoxygenation
pyroptosis
