1-磷酸鞘氨醇(S1P)对H9c2心肌细胞肥大反应的保护作用  

Protective effects of Sphingosine-1-phosphate(S1P)on hypertrophic response in H9c2 cardiomyocytes

作　　者：严惠 赵虎[2] 李论[1] YAN Hui;ZHAO Hu;LI Lun(Wuhan Fourth Hospital,Puai Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430000;Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430000,China)

机构地区：[1]武汉市第四医院华中科技大学同济医学院附属普爱医院,湖北武汉430000 [2]华中科技大学同济医学院附属同济医院,湖北武汉430000

出　　处：《中国应用生理学杂志》2022年第5期510-514,共5页Chinese Journal of Applied Physiology

摘　　要：目的:研究1-磷酸鞘氨醇(S1P)对H9c2心肌细胞肥大反应的影响。方法:将培养的H9c2心肌细胞随机分为4组,即正常对照组、S1P(1μmol/L)处理组、苯肾上腺素(PE,100μmol/L)处理组、PE(100μmol/L)加S1P(1μmol/L)处理组,每组设3个复孔。处理24 h后应用Actin-Trakcer Green免疫荧光染色检测各组心肌细胞形态大小;Real-Time PCR技术测定各组H9c2心肌细胞中肥大标志物ANP、BNP及β-MHC的转录水平;Western印迹法测定各组中ANP的蛋白表达情况。然后将H9c2心肌细胞随机分为5组,即正常对照组、PE(100μmol/L)组、PE(100μmol/L)加低浓度S1P(0.1μmol/L)组、PE加中浓度S1P(1μmol/L)组、PE加高浓度S1P(10μmol/L)组,每组设3个复孔。处理24 h后应用Western印迹法测定低、中、高浓度S1P干预下磷酸化的Janus激酶2(JAK2)及信号转导和转录激活子3(STAT3)的蛋白表达水平。每项试验独立重复三次。结果:与正常对照组比较,PE组的H9c2心肌细胞表面积显著增大(P<0.05),ANP、BNP及β-MHC的转录水平显著升高(P均<0.05),ANP的表达亦显著升高(P<0.05),而与PE组比较,PE加S1P组的H9c2心肌细胞表面积显著减小(P<0.05),ANP、BNP及β-MHC的转录水平显著降低(P均<0.05),ANP的表达亦显著降低(P<0.05);在PE加不同浓度S1P处理后,与正常对照组及PE组相比,p-JAK2和p-STAT3表达均显著升高(P<0.05),且呈一定的剂量依赖性。结论:S1P可以减轻PE诱导的心肌细胞肥大反应,这一保护作用可能与JAK2/STAT3信号通路的激活相关。Objective:To investigate the effects of sphingosine-1-phosphate(S1P)on cardiac hypertrophic response in H9c2 cells.Methods:H9c2 cells were randomly divided into four groups:normal control group,S1P(1μmol/L)treated group,Phenylephrine(PE)(100μmol/L)treated group,PE(100μmol/L)treated group combined with S1P(1μmol/L)treatment.Each group has 3 duplicated wells.After 24 hours,the size of H9c2 cells in each group was detected by Actin-Trakcer Green immunofluorescence staining.Transcriptional levels of hypertrophic markers(ANP,BNP andβ-MHC)in H9c2 cells were determined by real-time PCR.Western blot was performed to examine the expression level of ANP in each group.Then H9c2 cells were randomly divided into five groups:normal control group,PE(100μmol/L)treated group,PE(100μmol/L)with S1P low-dose(0.1μmol/L)treated group,PE(100μmol/L)with S1P middle-dose(1μmol/L)treated group and PE(100μmol/L)with S1P high-dose(10μmol/L)treated group.Each group has 3 duplicated wells.After 24 hours,Western blot was performed to examine the expressions of phosphorylated Janus kinase 2(JAK2)and signal transducers and activators of transcription 3(STAT3)under low,medium and high concentrations of S1P.Each experiment was repeated three times.Results:Compared with normal control group,the surface area of H9c2 cells in PE group was increased significantly(P<0.05),meanwhile,the transcription levels of ANP,BNP andβ-MHC were increased significantly(all P<0.05),and the expression of ANP was also increased significantly(P<0.05)in PE group.While compared with PE group,the surface area of H9c2 cells in PE+S1P group was decreased significantly(P<0.05),the transcription levels of ANP,BNP andβ-MHC and the expression of ANP were also decreased significantly(all P<0.05)in PE+S1P group.After treated with PE and different concentrations of S1P,the expressions of p-JAK2 and p-STAT3 were increased significantly compared with the normal control group and PE group(P<0.05),in a dose-dependent manner.Conclusion:S1P could protect H9c2 cells against hy

关 键 词：1-磷酸鞘氨醇 心肌细胞 肥大反应 Janus激酶2/信号转导和转录激活子3 细胞培养 

分 类 号：R541[医药卫生—心血管疾病]