摘要
目的:探讨硫化氢(H2S)在缺氧诱导的肺动脉平滑肌细胞(PASMCs)增殖、迁移和凋亡中的作用和可能的作用机制。方法:PASMCs随机分为三组,常氧对照组:给予常规条件下培养,低氧模型组:给与低氧(O2浓度3%)培养,建立细胞缺氧模型,硫氢化钠(NaHS)干预组:给与低氧联合硫氢化钠NaHS培养,培养24 h后,分别用CCK-8、流式细胞术、伤口愈合实验检测各组细胞增殖、凋亡、迁移,WB检测增殖、凋亡、迁移相关蛋白以及缺氧诱导因子1ɑ(HIF-1ɑ)和沉默信息调节因子2相关酶1(SIRT1)蛋白表达。结果:与常氧对照组比较,低氧模型组细胞增殖率显著升高,凋亡率显著降低,迁移能力上升,CDK2、CyclinD1、Bcl-2、基质金属蛋白酶-2(MMP-2)、MMP-9和HIF-1ɑ蛋白表达显著升高,Bax、SIRT1蛋白表达显著降低(P<0.05);而与低氧模型组比较,NaHS干预组细胞增殖率显著降低,凋亡率显著升高,迁移能力下降,CDK2、CyclinD1、Bcl-2、MMP-2、MMP-9和HIF-1ɑ蛋白表达显著下降,Bax、SIRT1蛋白表达显著升高(P<0.05)。结论:缺氧条件可以诱导PASMCs细胞增殖、迁移,抑制凋亡,抑制SIRT1表达,而硫化氢可以逆转缺氧诱导的增殖、迁移,促进凋亡,并上调SIRT1表达。
Objective: To investigate the effects and mechanism of hydrogen sulfide(H2S) on proliferation, migration and apoptosis of pulmonary arterial smooth muscle cells(PASMCs) induced by hypoxia. Methods: PASMCs were randomly divided into three groups. Firstly, the normal oxygen control group was cultured under normal conditions. Secondly, the hypoxia model group was cultured with 3% O2 concentration to establishment of a cell hypoxia model. Finally, administration sodium hydrosulfide(NaHS) combined with hypoxia as NaHS group. After 24 hours of culture, the cell proliferation, apoptosis, and migration were measured by CCK-8, flow cytometry, and wound healing experiments. Expression of proliferation, apoptosis and migration-related proteins, hypoxia inducible factor-1ɑ(HIF-1ɑ) and deacetylase SIRT1 protein were detected by western blot(WB). Results: Compared with normal oxygen control group, the cell proliferation and migration in the hypoxia model group were significantly increased, the apoptosis rate was significantly reduced. The expression of CDK2, CyclinD1, Bcl-2, and matrix metalloproteinase-2(MMP-2), MMP-9 and HIF-1α were increased, while the expression of Bax and SIRT1 were decreased(P<0.05). Compared with the hypoxia model group, the cell proliferation and migration in the NaHS group were significantly decreased, the apoptosis rate was increased. The expressions of CDK2, CyclinD1, Bcl-2, MMP-2, MMP-9, and HIF-1α were significantly reduced, and the expressions of Bax and SIRT1 were significantly increased(P<0.05). Conclusions: Hypoxia can induce the proliferation, migration and inhibit apoptosis of PASMCs cells and inhibit the expression of SIRT1, while H2 S can reverse the proliferation, migration, and promote apoptosis, and up regulate SIRT1 expression.
作者
曾闵
易世江
王野峰
杨舟
王祥
向金星
左超
陈智
ZENG Min;YI Shijiang;WANG Yefeng;YANG Zhou;WANG Xiang;XIANG Jinxing;ZUO Chao;CHEN Zhi(Hunan Children’s Hospital Cardiology,Changsha 410007,China)
出处
《心肺血管病杂志》
2020年第10期1258-1263,共6页
Journal of Cardiovascular and Pulmonary Diseases
关键词
硫化氢
沉默信息调节因子2相关酶1
肺动脉平滑肌细胞
增殖
迁移
凋亡
Hydrogen sulfide
Silent information regulator 2 related enzyme 1
Pulmonary artery smooth muscle cells
Proliferation
Migration
Apoptosis
