中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2015年
1期
45-49
,共5页
张帅%刘波%张波%樊再雯
張帥%劉波%張波%樊再雯
장수%류파%장파%번재문
肌细胞,平滑肌%缺氧%生存素%凋亡
肌細胞,平滑肌%缺氧%生存素%凋亡
기세포,평활기%결양%생존소%조망
Myocytes,smooth muscle%Anoxia%Survivin%Apoptosis
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.
目的 探討生存素在缺氧人肺動脈平滑肌細胞(HPASMCs)中的錶達情況及生存素的小分子抑製劑YM155對HPASMCs增殖與凋亡的影響.方法 將HPASMCs進行常氧或缺氧24 h培養併將其分為6組:常氧對照組(N組);24 h缺氧組(H組);常氧+YM155 10 nmol/L組(NY組);24 h缺氧+YM155 l nmol/L組(HY1組);24 h缺氧+YM155 10 nmol/L組(HY10組);24 h缺氧+YM155 100 nmol/L組(HY100組).採用Western blot法檢測HPASMCs生存素蛋白錶達,RT-PCR檢測HPASMCs生存素mRNA錶達.採用活細胞計數(CCK-8)法檢測HPASMCs增殖活性(吸光度值,A值),原位末耑標記(TUNEL)法檢測HPASMCs凋亡率.結果 N組未見生存素蛋白(0.016±0.006)和mRNA錶達,H組可見生存素蛋白(0.837±0.027)和生存素mRNA(17 086±1 044)錶達.各劑量YM155榦預組生存素蛋白、生存素mRNA含量分彆為0.382±0.041、0.281 ±0.025、0.021±0.002、8 074 ±2 135、5 614±709、1 382±347,較H組顯著降低(q值分彆為20.26、24.77、36.36、8.59、11.14、15,53,均P＜0.05),且在一定範圍內呈現濃度依賴性.N組細胞增殖活性和細胞凋亡率分彆為0.99±0.07、2.68±1.36,與H組(1.44±0.12、0.61 ±0.50)比較差異有統計學意義(q值分彆為6.484、3.532,均P＜0.05).各劑量YM155榦預組細胞增殖活性和細胞凋亡率分彆為1.32±0.07、1.17±0.07、0.84 ±0.13、5.52±1.71、6.66±1.49、7.97±1.93,與H組比較差異有統計學意義(g值分彆為2.581、3.980、8.733、6.014、7、413、9.023,均P＜0.05),且在一定範圍內呈現濃度依賴性.結論 生存素在缺氧性HPASMCs中錶達,但在常氧性HPASMCs中不錶達.生存素的小分子抑製劑YM155可以促進HPASMCs凋亡,抑製其增殖.生存素的錶達異常在缺氧性肺動脈高壓(HPH)的髮生髮展中髮揮重要作用,可能成為治療HPH的新靶點.
목적 탐토생존소재결양인폐동맥평활기세포(HPASMCs)중적표체정황급생존소적소분자억제제YM155대HPASMCs증식여조망적영향.방법 장HPASMCs진행상양혹결양24 h배양병장기분위6조:상양대조조(N조);24 h결양조(H조);상양+YM155 10 nmol/L조(NY조);24 h결양+YM155 l nmol/L조(HY1조);24 h결양+YM155 10 nmol/L조(HY10조);24 h결양+YM155 100 nmol/L조(HY100조).채용Western blot법검측HPASMCs생존소단백표체,RT-PCR검측HPASMCs생존소mRNA표체.채용활세포계수(CCK-8)법검측HPASMCs증식활성(흡광도치,A치),원위말단표기(TUNEL)법검측HPASMCs조망솔.결과 N조미견생존소단백(0.016±0.006)화mRNA표체,H조가견생존소단백(0.837±0.027)화생존소mRNA(17 086±1 044)표체.각제량YM155간예조생존소단백、생존소mRNA함량분별위0.382±0.041、0.281 ±0.025、0.021±0.002、8 074 ±2 135、5 614±709、1 382±347,교H조현저강저(q치분별위20.26、24.77、36.36、8.59、11.14、15,53,균P＜0.05),차재일정범위내정현농도의뢰성.N조세포증식활성화세포조망솔분별위0.99±0.07、2.68±1.36,여H조(1.44±0.12、0.61 ±0.50)비교차이유통계학의의(q치분별위6.484、3.532,균P＜0.05).각제량YM155간예조세포증식활성화세포조망솔분별위1.32±0.07、1.17±0.07、0.84 ±0.13、5.52±1.71、6.66±1.49、7.97±1.93,여H조비교차이유통계학의의(g치분별위2.581、3.980、8.733、6.014、7、413、9.023,균P＜0.05),차재일정범위내정현농도의뢰성.결론 생존소재결양성HPASMCs중표체,단재상양성HPASMCs중불표체.생존소적소분자억제제YM155가이촉진HPASMCs조망,억제기증식.생존소적표체이상재결양성폐동맥고압(HPH)적발생발전중발휘중요작용,가능성위치료HPH적신파점.
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 nmol/L YM155 groups (5.52 ± 1.71,6.66 ± 1.49,7.97 ± 1.93,q =6.014,7.413,9.023,all P ＜0.05) enhanced in a dose-dependent manner.Conclusions Survivin was expressed in hypoxic HPASMCs,but not in normal HPASMCs.Treatment with the survivin inhibitor YM155 led to decreased proliferation and enhanced apoptosis in hypoxic HPASMCs.Survivin might be a significant target for human hypoxic pulmonary hypertension.
