Wu Yang, Guo Yuan-Yuan, Zhang Yuan-Yuan, Zhang Yi
Institute of Navigation Medicine, Nantong University, Nantong 226019, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2018 Jan 8;34(1):29-34. doi: 10.12047/j.cjap.5492.2018.009.
To investigate the effects of hydrogen sulfide (HS) on the negatively regulation of cardiomyocyte hypertrophy and the relationship between the effect of HS with miRNA-133a-mediated Ca/calcineurin/NFATc4 signal pathway.
Cardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (Leica). The expression of brain natriuretic peptide(BNP), β-myosin heavy chain(β-MHC), cystathionase (CSE), miRNA-133a, calcineurin (CaN) were detected by qRT-PCR. The protein expressions of CaN、nuclear factors of activated T cells (NFATc4) were detected by Western blot. The concentration of HS in the cardiomyocyte was detected by Elisa. The concentration of intracellular calcium was measured by calcium imaging using confocal microscope. The nuclear translocation of NFATc4 was checked by immuno-fluorescence cell staining technique.
①The level of system of CSE/HS and expression of miRNA-133a were significantly reduced in cardiomyocyte hypertrophy. Pretreatment with NaHS increased the concentration of HS and the expression of miRNA-133a mRNA in cardiomyocytes, and suppressed cardiomyocyte hypertrophy. ②The concentration of intracellular calcium, the expression of CaN and nulear protein NFATc4 were significantly increased, and the nuclear translocation of NFATc4 were obviously enhanced in cardiomyocyte hypertrophy. NaHS pretreatment markedly inhibited these effects of ISO induced cardiomyocyte hypertrophy. ③Application of antagomir-133a reversed the inhibitory effects of NaHS on cardiomyocyte hypertrophy, and increased the influx of intracellular calcium, and elevated the expression of CaN and nuclear protein NFATc4, and enhanced the nuclear translocation of NFATc4.
HS can negatively regulate cardiomyocyte hypertrophy. The effects might be associated with HS increasing expression of miRNA-133a and inhibiting inactivation of Ca/calcineurin/NFATc4 signal pathway.
探讨硫化氢（HS）对心肌细胞肥大负向调控的作用以及HS作用与miRNA - 133a介导的Ca/钙调神经磷酸酶/NFATc4信号通路之间的关系。
采用异丙肾上腺素（ISO）诱导心肌细胞肥大。通过图像分析系统（徕卡）测量细胞表面积。采用qRT - PCR检测脑钠肽（BNP）、β - 肌球蛋白重链（β - MHC）、胱硫醚γ-裂解酶（CSE）、miRNA - 133a、钙调神经磷酸酶（CaN）的表达。采用蛋白质免疫印迹法检测CaN、活化T细胞核因子（NFATc4）的蛋白表达。采用酶联免疫吸附测定法（Elisa）检测心肌细胞中HS浓度。使用共聚焦显微镜通过钙成像测量细胞内钙浓度。采用免疫荧光细胞染色技术检测NFATc4的核转位。
①在心肌细胞肥大中，CSE/HS系统水平和miRNA - 133a表达显著降低。用硫氢化钠（NaHS）预处理可增加心肌细胞中HS浓度和miRNA - 133a mRNA表达，并抑制心肌细胞肥大。②在心肌细胞肥大中，细胞内钙浓度、CaN表达及核蛋白NFATc4显著增加，NFATc4的核转位明显增强。NaHS预处理显著抑制ISO诱导的心肌细胞肥大的这些效应。③应用抗miR - 133a可逆转NaHS对心肌细胞肥大的抑制作用，增加细胞内钙内流，提高CaN和核蛋白NFATc4的表达，并增强NFATc4的核转位。
HS可负向调控心肌细胞肥大。其作用可能与HS增加miRNA - 133a表达及抑制Ca/钙调神经磷酸酶/NFATc4信号通路失活有关。
