1-磷酸鞘氨醇(S1P)对H9c2心肌细胞肥大反应的保护作用*

doi:10.12047/j.cjap.6281.2022.095

摘要/Abstract

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(1 μmol/L)处理组、苯肾上腺素(PE,100 μmol/L)处理组、PE(100 μmol/L)加S1P(1 μmol/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(100 μmol/L)组、PE(100 μmol/L)加低浓度S1P(0.1 μmol/L)组、PE加中浓度S1P(1 μmol/L)组、PE加高浓度S1P(10 μmol/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。

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

Abstract: 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 (1 μmol/L) treated group, Phenylephrine (PE) (100 μmol/L) treated group, PE (100 μmol/L) treated group combined with S1P (1 μmol/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 and β-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 (100 μmol/L) treated group, PE (100 μmol/L) with S1P low-dose (0.1 μmol/L) treated group, PE (100 μmol/L) with S1P middle-dose (1 μmol/L) treated group and PE (100 μmol/L) with S1P high-dose (10 μmol/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 and β-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 and β-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 hypertrophic response induced by PE, which may be achieved by activating JAK2/STAT3 signal pathway.

Key words: sphingosine-1-phosphate, cardiomyocytes, hypertrophic response, JAK2/STAT3, cell culture

中图分类号:

R541

严惠, 赵虎, 李论. 1-磷酸鞘氨醇(S1P)对H9c2心肌细胞肥大反应的保护作用^*[J]. 中国应用生理学杂志, 2022, 38(5): 510-514.

YAN Hui, ZHAO Hu, LI Lun. Protective effects of Sphingosine-1-phosphate (S1P) on hypertrophic response in H9c2 cardiomyocytes[J]. CJAP, 2022, 38(5): 510-514.

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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

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