Objective:To explore the relationship between hypoxic pulmonary arterial smooth muscle cells(PASMCs)proliferation,apoptosis and mitogen-activated protein kinases(MAPK) signal pathway in rats.Methods:PASMCs were obtained from male SD rats by the enzyme digestion method and primarily cultured;PASMCs were identified through two methods:immunofluorescence staining and light microscopy;the 4 ～ 6th generation PASMCs of logarithmic growth state of good growth period were selected,and randomly divided into 7 groups:normoxic control group (N),hypoxia group (H),DMSO group (D),extracellular signal-regulated kinase1/2(ERK1/2) inhibitor-U0126 group (U) and p38MAPK inhibitor-SB203580 group (S),the p38MAPK activator-Anisomycin group (A),the ERK1/2 activator-Staurosporine Aglycone group (SA).When all the models were completed,the all groups joined the CCK-8 to measure cell proliferation;cell apoptosis of each group was detected by TUNEL kit after the modeling.Results:Compared with N group,the expression of OD value in H group was up-regulated (0.990 ± 0.041 vs 1.143 ± 0.033,P ＜ 0.01).There was no statistical significance on PASMCs apoptosis index(AI) in H group (4.913 ± 0.451 vs 5.452 ± 0.557,P ＞ 0.05);Compared With H group,there were no statistical significance on the expression of PASMCs OD value and apoptosis index(AI)in D group (1.143 ± 0.033 vs 1.142 ± 0.049,5.452 ± 0.557 vs 5.402 ± 0.651,P ＞ 0.05);the expression of OD value in U group was down-regulated,and the expression of AI was up-regulated (1.143 ± 0.033 vs 0.985 ± 0.078,5.452 ± 0.557 vs 10.145 ± 2.545,P ＜ 0.01);the expression of OD value in S group was up-regulated,and the expression of AI was down-regulated (1.143 ± 0.033 vs 1.295 ± 0.039,5.452 ± 0.557 vs 3.093 ± 0.409,P ＜ 0.01);the expression of OD value in A group was down-regulated,and the expression of AI was up-regulated (1.143 ± 0.033 vs 0.347 ± 0.067,5.452 ± 0.557 vs 25.753 ± 1.262,P ＜ 0.01);the expression of OD value in SA group was up-regulated,and the expression of AI was down-regulated (1.143 ± 0.033 vs 1.685 ± 0.100,5.452 ± 0.557 vs 1.700 ±0.095,P ＜ 0.01).Conclusion:The regulation of PASMCs' proliferation and apoptosis under hypoxia condition have a relationship with the participation of MAPK signal pathway.%目的:研究低氧性大鼠肺动脉平滑肌细胞(PASMC)的增殖、凋亡与丝裂原活化蛋白激酶(MAPK)关系.方法:用组织酶消化法获取肺动脉平滑肌细胞(PASMCs),进行原代培养;采用普通光学显微镜和免疫荧光染色法,分别鉴定PASMCs;选择处于对数生长期的4～6代PASMCs,随机分为7组进行造模:常氧对照组(N)、低氧组(H)、DM-SO组(D)、U0126组(U)、SB203580组(S)、Anisomycin组(A)、Staurosporine Aglycone组(SA);N组加入10％培养基后置于常氧培养箱中,其它各组分别加入含相应药物的10％培养基后置于低氧培养箱(3％O2,5％CO2,37℃)中,造模时间均为48 h.CCK-8法检测各组PASMCs增殖情况;TUNEL法测定各组PASMCs凋亡情况.结果:与N组相比,H组PASMCs的OD值显著上调(0.990±0.041 vs 1.143±±0.033,P＜0.01),凋亡指数没有明显变化(4.913±0.451 vs5.452± 0.557,P＞0.05);与H组相比,D组PASMCs的OD值和凋亡指数均无显著变化(1.143±0.033 vs 1.142 ±0.049,5.452±0.557 us 5.402± 0.651,均P＞0.05);U组PASMCs的OD值下降,凋亡指数升高(1.143± 0.033 vs0.985± 0.078,5.452±0.557 vs 10.145± 2.545,均P＜0.01);S组PASMCs OD值上调,凋亡指数明显下调(1.143±0.033 vs 1.295± 0.039,5.452±0.557 vs 3.093±0.409,均P＜0.01);A组PASMCs的OD值下降,凋亡指数升高(1.143± 0.033 vs 0.347±0.067,5.452±0.557 vs 25.753±1.262,均P＜0.01);SA组PASMCs OD值上调,凋亡指数下调(1.143±0.033 vs 1.685±0.100,5.452±0.557 vs 1.700± 0.095,均P＜0.01).结论:低氧对PASMCs增殖和凋亡的调控与MAPK信号通路有关.
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