中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
9期
1645-1650
,共6页
马迎春%郑梦晓%黄林静%王园园%应磊%王万铁
馬迎春%鄭夢曉%黃林靜%王園園%應磊%王萬鐵
마영춘%정몽효%황림정%왕완완%응뢰%왕만철
Kv1 .5钾通道%丝裂原激活蛋白激酶%肺动脉平滑肌细胞%细胞增殖%细胞凋亡
Kv1 .5鉀通道%絲裂原激活蛋白激酶%肺動脈平滑肌細胞%細胞增殖%細胞凋亡
Kv1 .5갑통도%사렬원격활단백격매%폐동맥평활기세포%세포증식%세포조망
Potassium channels%Mitogen-activated protein kinase%Pulmonary artery smooth muscle cells%Cell proliferation%Apoptosis
目的:探讨电压依赖性钾离子通道Kv1.5对大鼠低氧高二氧化碳性肺动脉平滑肌细胞( PASMCs)增殖、凋亡的影响及其与丝裂原激活蛋白激酶( MAPK)信号通路的关系。方法:体外培养大鼠PASMCs,复制低氧高二氧化碳模型,随机分组如下:常氧组( N组);低氧高二氧化碳组( HH组);低氧高二氧化碳+溶剂DMSO对照组( HD组);低氧高二氧化碳+ERK1/2通路抑制剂U0126组( HU组);低氧高二氧化碳+p38 MAPK通路抑制剂SB203580组( HS组);低氧高二氧化碳+MAPK通路激动剂茴香霉素( anisomycin )组( HA组)。采用CCK-8法检测细胞活性,蛋白免疫印迹法检测Kv1.5、增殖细胞核抗原( PCNA)及Bax蛋白的表达水平。结果:与N组相比,HH组和HD组细胞活性增加(P<0.01),PCNA蛋白表达上调,Kv1.5及Bax蛋白表达均明显降低(P<0.01), HH组和HD组间各指标变化均无显著差异(均P>0.05);较之HD组,HU组、HS组及HA组细胞活性降低( P<0.05或P<0.01),PCNA蛋白表达下调,Kv1.5及Bax蛋白表达均明显增加,差异均显著(P<0.01),其中以HA组各指标变化最明显。结论:钾离子通道Kv1.5对低氧高二氧化碳性大鼠PASMCs增殖、凋亡的调节可能与MAPK通路的激活有关。
目的:探討電壓依賴性鉀離子通道Kv1.5對大鼠低氧高二氧化碳性肺動脈平滑肌細胞( PASMCs)增殖、凋亡的影響及其與絲裂原激活蛋白激酶( MAPK)信號通路的關繫。方法:體外培養大鼠PASMCs,複製低氧高二氧化碳模型,隨機分組如下:常氧組( N組);低氧高二氧化碳組( HH組);低氧高二氧化碳+溶劑DMSO對照組( HD組);低氧高二氧化碳+ERK1/2通路抑製劑U0126組( HU組);低氧高二氧化碳+p38 MAPK通路抑製劑SB203580組( HS組);低氧高二氧化碳+MAPK通路激動劑茴香黴素( anisomycin )組( HA組)。採用CCK-8法檢測細胞活性,蛋白免疫印跡法檢測Kv1.5、增殖細胞覈抗原( PCNA)及Bax蛋白的錶達水平。結果:與N組相比,HH組和HD組細胞活性增加(P<0.01),PCNA蛋白錶達上調,Kv1.5及Bax蛋白錶達均明顯降低(P<0.01), HH組和HD組間各指標變化均無顯著差異(均P>0.05);較之HD組,HU組、HS組及HA組細胞活性降低( P<0.05或P<0.01),PCNA蛋白錶達下調,Kv1.5及Bax蛋白錶達均明顯增加,差異均顯著(P<0.01),其中以HA組各指標變化最明顯。結論:鉀離子通道Kv1.5對低氧高二氧化碳性大鼠PASMCs增殖、凋亡的調節可能與MAPK通路的激活有關。
목적:탐토전압의뢰성갑리자통도Kv1.5대대서저양고이양화탄성폐동맥평활기세포( PASMCs)증식、조망적영향급기여사렬원격활단백격매( MAPK)신호통로적관계。방법:체외배양대서PASMCs,복제저양고이양화탄모형,수궤분조여하:상양조( N조);저양고이양화탄조( HH조);저양고이양화탄+용제DMSO대조조( HD조);저양고이양화탄+ERK1/2통로억제제U0126조( HU조);저양고이양화탄+p38 MAPK통로억제제SB203580조( HS조);저양고이양화탄+MAPK통로격동제회향매소( anisomycin )조( HA조)。채용CCK-8법검측세포활성,단백면역인적법검측Kv1.5、증식세포핵항원( PCNA)급Bax단백적표체수평。결과:여N조상비,HH조화HD조세포활성증가(P<0.01),PCNA단백표체상조,Kv1.5급Bax단백표체균명현강저(P<0.01), HH조화HD조간각지표변화균무현저차이(균P>0.05);교지HD조,HU조、HS조급HA조세포활성강저( P<0.05혹P<0.01),PCNA단백표체하조,Kv1.5급Bax단백표체균명현증가,차이균현저(P<0.01),기중이HA조각지표변화최명현。결론:갑리자통도Kv1.5대저양고이양화탄성대서PASMCs증식、조망적조절가능여MAPK통로적격활유관。
AIM:To investigate the effects of voltage-dependent K +channel 1.5 (Kv1.5) on the proliferation and apoptosis of rat pulmonary artery smooth muscle cells (PASMCs) under hypoxia+hypercapnia condition and the relation-ship with mitogen-activated protein kinase(MAPK) signal pathway.METHODS:The PASMCs isolated from the male SD rat were cultured under hypoxia +hypercapnia condition, and randomly divided into normal group (N group), hypoxia+hyper-capnia group (HH group), hypoxia+hypercapnia+DMSO incubation group (HD group), hypoxia+hypercapnia +U0126 (an extracellular signal-regulated kinase 1/2 inhibitor) incubation group (HU group), hypoxia+hypercapnia+SB203580 (a p38 mitogen-activated protein kinase inhibitor ) incubation group (HS group), and hypoxia+hypercapnia+anisomycin (an agonist of MAPK) incubation group (HA group).Cell Counting Kit-8 was used to detect the cell viability.The protein expression of Kv1.5, PCNA and Bax was detected by Western blotting .RESULTS:Compared with N group , the cell via-bility and PCNA protein expression in HH group and HD group were significantly raised (P<0.01), but Kv1.5 and Bax proteins were significantly decreased (P<0.01).No difference between HH group and HD group was observed (P>0.05 ) .Compared with HD group , the cell viability and PCNA protein expression in HU group , HS group and HA group were decreased (P<0.05 or P<0.01), but Kv1.5 protein and Bax protein were raised (P<0.01), with the most significant changes in HA group .CONCLUSION:The regulation of Kv1.5 to the proliferation and apoptosis of PASMCs under hy-poxia+hypercapnia condition might have a relationship with the activation of MAPK signal pathway .