MAPK信号通路对大鼠低氧性PASMCs增殖、凋亡的调控

doi:10.12047/j.cjap.5422.2017.056

中国应用生理学杂志 ›› 2017, Vol. 33 ›› Issue (3): 226-230.doi: 10.12047/j.cjap.5422.2017.056

MAPK信号通路对大鼠低氧性PASMCs增殖、凋亡的调控

黄林静^1,2, 张聪聪¹, 赵美平¹, 郑梦晓¹, 应磊¹, 陈锡文³, 王万铁¹

1. 温州医科大学病理生理学教研室, 浙江温州 325035;
2. 温州医科大学附属第二医院病理科, 浙江温州 325035;
3. 温州医科大学实验动物中心, 浙江温州 325035

收稿日期:2016-02-01 修回日期:2017-01-16 出版日期:2017-05-28 发布日期:2018-06-20
通讯作者: 王万铁,Tel:0577-86689817,E-mail:wwt@wmu.edu.cn E-mail:wwt@wmu.edu.cn
基金资助:
浙江省温州市对外科技合作项目（HZ0090066）；浙江省公益性技术应用研究计划项目资助的课题（2015C37121）

The regulation of MAPK signaling pathway on cell proliferation and apoptosis in hypoxic PASMCs of rats

HUANG Lin-jing^1,2, ZHANG Cong-cong¹, ZHAO Mei-ping¹, ZHENG Meng-xiao¹, YING lei¹, CHEN Xi-wen³, WANG Wan-tie¹

1. Department of Pathophysiology, Wenzhou Medical University, Wenzhou 325035, China;
2. The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325035, China;
3. Experimental Animal Center, Wenzhou Medical University, Wenzhou 325035, China

Received:2016-02-01 Revised:2017-01-16 Online:2017-05-28 Published:2018-06-20
Supported by:
浙江省温州市对外科技合作项目（HZ0090066）；浙江省公益性技术应用研究计划项目资助的课题（2015C37121）

摘要/Abstract

摘要： 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 vs 5.452 ±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 vs 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 vs 0.985 ±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。

关键词: 低氧, 肺动脉平滑肌细胞, p38MAPK, ERK1/2, 大鼠, 增值, 凋亡

Abstract: Objective: To explore the relationship between hypoxic pulmonary arterial smooth muscle cells(PASMCs)proliferation, apop-tosis 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 con-trol 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 expres-sion 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.

Key words: hypoxia, PASMCs, p38MAPK, ERK1/2, rat, proliferation, apoptosis

黄林静, 张聪聪, 赵美平, 郑梦晓, 应磊, 陈锡文, 王万铁. MAPK信号通路对大鼠低氧性PASMCs增殖、凋亡的调控[J]. 中国应用生理学杂志, 2017, 33(3): 226-230.

HUANG Lin-jing, ZHANG Cong-cong, ZHAO Mei-ping, ZHENG Meng-xiao, YING lei, CHEN Xi-wen, WANG Wan-tie. The regulation of MAPK signaling pathway on cell proliferation and apoptosis in hypoxic PASMCs of rats[J]. CJAP, 2017, 33(3): 226-230.

参考文献

[1] Jin N, Hatton N, Swartz DR, et al. Hypoxia activates jun-N-terminal kinase, extracellular signal-regulated protein ki-nase, and p38kinase in pulmonary arteries[J]. Am J Respir Cell Mol Biol, 2000, 23(5):593-601.
[2] Stenmark KR, Meyrick B, Galie N, et al. Animal models of pulmonary arterial hypertension:the hope for etiological dis-covery and pharmacological cure[J]. Am J Physiol Lung Cell Mol Physiol, 2009, 297(6):L1013-1032.
[3] 王园园, 郑梦晓, 赵美平, 等. ERK1/2对低氧大鼠肺动脉平滑肌细胞Kv1.5通道表达的影响[J]. 中国应用生理学杂志, 2015, 31(5):418-421.
[4] Stenmark KR, Rabinovitch M. Emerging therapies for the treatment of pulmonary hypertension[J]. Pediatr Crit Care Med, 2010, 11(2Suppl):S85-S90.
[5] Vanderpool RR, Kim AR, Molthen R, et al. Effects of acute Rho kinase inhibition on chronic hypoxia-induced changes in proximal and distal pulmonary arterial structure and function[J]. J Appl Physiol (1985), 2011, 110(1):188-198.
[6] 朱焕勉, 陈然, 薛峰, 等. 自噬抑制剂氯奎对低氧诱导肺动脉平滑肌细胞增殖的影响[J]. 中国应用生理学杂志, 2014, 30(1):8-12.
[7] 范小芳, 李文娟, 陈兆琴, 等. 慢性低氧性肺动脉高压大鼠肺组织内质网应激介导的凋亡的变化[J]. 中国应用生理学杂志, 2011, 27(3):270-274.
[8] Liu Y, Zhao Y, Ju S, et al. Orexin A upregulates the pro-tein expression of OX1R and enhances the proliferation of SGC-7901gastric cancer cells through the ERK signaling pathway[J]. Int J Mol Med, 2015, 35(2):539-545.
[9] Li T, Song T, Ni L, et al. The p-ERK-p-c-Jun-cyclin D1 pathway is involved in proliferation of smooth muscle cells af-ter exposure to cigarette smoke extract[J]. Biochem Biophys Res Commun, 2014, 453(3):316-320.
[10] Chen YY, Hsieh CY, Jayakumar T, et al. Andrographolide induces vascular smooth muscle cell apoptosis through a SHP-1-PP2A-p38MAPK-p53 cascade[J]. Sci Rep, 2014, 4:5651.
[11] BAI J, LIU XS, XU YJ, et al. Regulation of cell prolifera-tion and apoptosis by ERK signaling pathway in airway smooth muscle cells of rats[J]. 中国生物化学与分子生物学报, 2007, 23(1):38-44.
[12] 卜祥宁, 韩松, 李俊发. 低氧预适应小鼠脑组织内P38MAPK磷酸化水平增高[J]. 基础医学与临床, 2007, 27(5):481-484.

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MAPK信号通路对大鼠低氧性PASMCs增殖、凋亡的调控

The regulation of MAPK signaling pathway on cell proliferation and apoptosis in hypoxic PASMCs of rats

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