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

doi:10.12047/j.cjap.6281.2022.095

Abstract

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

CLC Number:

R541

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.

References

[1] Sag CM, Santos CX, Shah AM. Redox regulation of cardiac hypertrophy [J]. J Mol Cell Cardiol, 2014, 73: 103-111.
[2] Gupta VK, You Y, Klistorner A, et al. Focus on molecules: Sphingosine 1 phosphate (S1P) [J]. Exp Eye Res, 2012, 103: 119-120.
[3] Jozefczuk E, Guzik TJ, Siedlinski M. Significance of sphingosine-1-phosphate in cardiovascular physiology and pathology [J]. Pharmacol Res, 2020, 156: 104793.
[4] Yao Mattisson I, Christoffersen C. Apolipoprotein M and its impact on endothelial dysfunction and inflammation in the cardiovascular system [J]. Atherosclerosis, 2021, 334: 76-84.
[5] Zhang J, Honbo N, Goetzl EJ, et al. Signals from type 1 sphingosine 1-phosphate receptors enhance adult mouse cardiac myocyte survival during hypoxia [J]. Am J Physiol Heart Circ Physiol, 2007, 293(5): H3150-3158.
[6] 袁磊, 张文杰, 张舵舵, 等. S1P对心肌细胞的保护作用研究 [J]. 中国应用生理学杂志, 2011, 27(3): 320-323.
[7] Wang Y, Wang D, Zhang L, et al. Role of JAK-STAT pathway in reducing cardiomyocytes hypoxia/reoxygenation injury induced by S1P postconditioning [J]. Eur J Pharmacol, 2016, 784: 129-136.
[8] 肖倩, 宝璐尔, 陈辰, 等. 1-磷酸鞘氨醇受体1型信号通路在压力超负荷诱导的心室重构中的作用及机制 [J]. 同济大学学报(医学版), 2018, 39(6): 46-53.
[9] Jensen BC, O'connell TD, Simpson PC. Alpha-1-adrenergic receptors in heart failure: the adaptive arm of the cardiac response to chronic catecholamine stimulation [J]. J Cardiovasc Pharmacol, 2014, 63(4): 291-301.
[10] 崔常贺, 陈芳, 徐予. H9c2细胞在心脏离体模型中研究进展 [J]. 中国实用医刊, 2014, 41(1): 92-94.
[11] 宋海岩, 周志新, 张玉祥. Notch3对促进胰腺星形细胞活化的基因表达及信号通路的影响 [J]. 中国应用生理学杂志, 2021, 37(4): 349-353.
[12] Shimizu I, Minamino T. Physiological and pathological cardiac hypertrophy [J]. J Mol Cell Cardiol, 2016, 97: 245-262.
[13] Sekiguchi K, Yokoyama T, Kurabayashi M, et al. Sphingosylphosphorylcholine induces a hypertrophic growth response through the mitogen-activated protein kinase signaling cascade in rat neonatal cardiac myocytes [J]. Circ Res, 1999, 85(11): 1000-1008.
[14] Robert P, Tsui P, Laville MP, et al. EDG1 receptor stimulation leads to cardiac hypertrophy in rat neonatal myocytes [J]. J Mol Cell Cardiol, 2001, 33(9): 1589-1606.
[15] Qi Y, Li JJ, Di XH, et al. Excess sarcoplasmic reticulum-mitochondria calcium transport induced by Sphingosine-1-phosphate contributes to cardiomyocyte hypertrophy [J]. Biochim Biophys Acta Mol Cell Res, 2021, 1868(5): 118970.
[16] Wei L. Lysophospholipid signaling in cardiac myocyte hypertrophy [J]. J Mol Cell Cardiol, 2004, 36(4): 465-468.
[17] Rong J, Li L, Jing L, et al. JAK2/STAT3 pathway mediates protection of metallothionein against doxorubicin-induced cytotoxicity in mouse cardiomyocytes [J]. Int J Toxicol, 2016, 35(3): 317-326.
[18] Shang L, Dong P, Du L, et al. SERP1 prevents hypoxia-reoxygenation-induced H9c2 apoptosis through activating JAK2/STAT3 pathway-dependent attenuation of endoplasmic reticulum stress [J]. Biochem Biophys Res Commun, 2019, 508(1): 256-262.
[19] Hilfiker-Kleiner D, Hilfiker A, Drexler H. Many good reasons to have STAT3 in the heart [J]. Pharmacol Ther, 2005, 107(1): 131-137.
[20] 孙晓娟, 冯武龙, 侯娜. JAK2/STAT3信号通路在运动预适应抗心肌细胞凋亡中的作用 [J]. 中国应用生理学杂志, 2017, 33(5): 393-397.
[21] Ke M, Tang Q, Pan Z, et al. Sphingosine-1-phosphate attenuates hypoxia/reoxygenation-induced cardiomyocyte injury via a mitochondrial pathway [J]. Biochem Biophys Res Commun, 2019, 510(1): 142-148.