生存素表达对缺氧人肺动脉平滑肌细胞凋亡与增殖的影响  

The effect of survivin expression on the apoptosis and proliferation of hypoxic human pulmonary arterial smooth muscle cells

作　　者：张帅[1] 刘波[1] 张波[2] 樊再雯[2] 

机构地区：[1]河北北方学院研究生部,张家口075000 [2]空军总医院呼吸内科

出　　处：《中华结核和呼吸杂志》2015年第1期45-49,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金（81170045）

摘　　要：目的 探讨生存素在缺氧人肺动脉平滑肌细胞（HPASMCs）中的表达情况及生存素的小分子抑制剂YM155对HPASMCs增殖与凋亡的影响.方法 将HPASMCs进行常氧或缺氧24 h培养并将其分为6组：常氧对照组（N组）;24 h缺氧组（H组）;常氧＋YM155 10 nmol/L组（NY组）;24 h缺氧＋YM155 l nmol/L组（HY1组）;24 h缺氧＋YM155 10 nmol/L组（HY10组）;24 h缺氧＋YM155 100 nmol/L组（HY100组）.采用Western blot法检测HPASMCs生存素蛋白表达,RT-PCR检测HPASMCs生存素mRNA表达.采用活细胞计数（CCK-8）法检测HPASMCs增殖活性（吸光度值,A值）,原位末端标记（TUNEL）法检测HPASMCs凋亡率.结果 N组未见生存素蛋白（0.016±0.006）和mRNA表达,H组可见生存素蛋白（0.837±0.027）和生存素mRNA（17 086±1 044）表达.各剂量YM155干预组生存素蛋白、生存素mRNA含量分别为0.382±0.041、0.281 ±0.025、0.021±0.002、8 074 ±2 135、5 614±709、1 382±347,较H组显著降低（q值分别为20.26、24.77、36.36、8.59、11.14、15,53,均P＜0.05）,且在一定范围内呈现浓度依赖性.N组细胞增殖活性和细胞凋亡率分别为0.99±0.07、2.68±1.36,与H组（1.44±0.12、0.61 ±0.50）比较差异有统计学意义（q值分别为6.484、3.532,均P＜0.05）.各剂量YM155干预组细胞增殖活性和细胞凋亡率分别为1.32±0.07、1.17±0.07、0.84 ±0.13、5.52±1.71、6.66±1.49、7.97±1.93,与H组比较差异有统计学意义（g值分别为2.581、3.980、8.733、6.014、7、413、9.023,均P＜0.05）,且在一定范围内呈现浓度依赖性.结论 生存素在缺氧性HPASMCs中表达,但在常氧性HPASMCs中不表达.生存素的小分子抑制剂YM155可以促进HPASMCs凋亡,抑制其增殖.生存素的表达异常在缺氧性肺动脉高压（HPH）的发生发展中发挥重要作用,可能成为治疗HPH的新靶点.Objective Survivin is a new member of the inhibitor of apoptosis （IAP） family of proteins,which is also the strongest inhibitor of apoptosis.It inhibits apoptosis and promotes cell proliferation.This study aims to explore the effect of survivin expression on the apoptosis and proliferation of hypoxic human pulmonary arterial smooth muscle cells （HPASMCs）.Methods HPASMCs were divided randomly into 6 groups：Normoxia （N group）：cultured under normoxia; Normoxia ＋ YM155 （NY group）：cultured under normoxia with YM155,a selective survivin inhibitor,for 24 h; Hypoxia（H group）：cultured under hypoxia for 24 h ; Hypoxia ＋ 1 nmol/L YM155 （HY1 group）：cultured under hypoxia with 1 nmol/L YM155 for 24 h; Hypoxia ＋ 10 nmol/L YM155 （HY10 group）：cultured under hypoxia with 10 nmoL/L YM155 for 24 h;Hypoxia ＋ 100 nmol/L YM155 （HY100 group）：cultured under hypoxia with 100 nmol/L YM155 for 24 h.The mRNA and protein expressions of survivin were measured by real time PCR and Western Blot respectively.Cell proliferation was determined using a Cell Counting Kit-8 （CCK-8）.Apoptosis was detected with a TUNEL test.Results The mRNA and protein expressions of survivin were observed in the H group （17 086 ±1 044,0.837 ±0.027）,but not in the N group（1,0.016 ±0.06）.The cell proliferation in H group （1.44 ±0.12） was significantly increased compared with that of the N group （0.99 ±0.07,q =6.484,P ＜ 0.05）.The apoptosis in the H group （0.61 ± 0.50） was significantly decreased compared with that in the N group （2.68 ± 1.36,q =3.532,P ＜ 0.05）.As compared with the H group,the survivin mRNA,survivin protein and cell proliferation in hypoxia plus 1 nmol/L,10 nmol/L,100 nmol/L YM155 groups （8 074 ± 2 135,5 614 ± 709,1 382 ± 347,0.382 ± 0.041,0.281 ± 0.025,0.021 ± 0.002,1.318 ±0.067,1.168 ±0.071,0.845 ±0.129,q =8.59,11.14,15.53,20.26,24.77,36.36,2.58,3.98,8.73,all P ＜ 0.05） significant decreased,but the apoptosis in hypoxia plus 1 nmol/L,10 nmol/L,100

关 键 词：肌细胞 平滑肌 缺氧 生存素 凋亡 

分 类 号：R544.1[医药卫生—心血管疾病]