内质网应激在大鼠低氧高二氧化碳性肺动脉高压中的作用

doi:10.12047/j.cjap.5644.2018.075

中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (4): 327-333.doi: 10.12047/j.cjap.5644.2018.075

内质网应激在大鼠低氧高二氧化碳性肺动脉高压中的作用

张晶晶¹, CHEN Jun-hao², 赵美平¹, 武垣伶¹, 张聪聪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. School of Biomedical Sciences, University of Western Australia, Perth 6000, Australia;
3. 温州医科大学实验动物中心, 浙江温州 325035

收稿日期:2017-09-28 修回日期:2018-02-01 出版日期:2018-07-28 发布日期:2019-02-21
通讯作者: 王万铁 E-mail:wwt@wmu.edu.cn
基金资助:
温州市高层次人才创新技术重点资助项目（2016-07）；浙江省中医药重点研究计划（2013ZZ011）

The role of endoplasmic reticulum stress in pulmonary hypertension in rat induced by chronic hypoxia and hypercapnia

ZHANG Jing-jing¹, CHEN Jun-hao², ZHAO Mei-ping¹, WU Yuan-ling¹, ZHANG Cong-cong¹, YING Lei¹, CHEN Xi-wen³, WANG Wan-tie¹

1. Department of Pathophysiology, Wenzhou Medical University, Wenzhou 325035;
2. School of Biomedical Sciences, University of Western Australia, Perth 6000, Australia;
3. Animal Experiment Center, Wenzhou Medical University, Wenzhou 325035, China

Received:2017-09-28 Revised:2018-02-01 Online:2018-07-28 Published:2019-02-21
Supported by:
温州市高层次人才创新技术重点资助项目（2016-07）；浙江省中医药重点研究计划（2013ZZ011）

摘要/Abstract

摘要： 目的：观察低氧高二氧化碳性肺动脉高压大鼠的肺血管重塑并探讨内质网应激（ERS）在肺动脉高压中的作用。方法：将40只SD大鼠随机分为四组：常氧对照组（N）、低氧高二氧化碳组（HH）、ERS通路抑制剂4-苯基丁酸（4-phenylbutyric acid）组（4-PBA）、ERS通路激动剂衣霉素（tunicamycin）组（TM），n=10。测量各组大鼠的肺动脉平均压（mPAP）、颈动脉平均压以及右心室肥大指数，免疫荧光α-SMA标记法鉴定各组肺中小动脉平滑肌细胞，电镜观察肺组织及肺中小动脉形态学变化，原位末端标记法（TUNEL）检测各组肺动脉平滑肌细胞的凋亡指数，采用RT-PCR和Western blot分别检测各组大鼠葡萄糖调节蛋白78（GRP78）、C/EBP同源蛋白（CHOP）、c-Jun氨基末端激酶（JNK）、天冬氨酸特异性半胱氨酸蛋白酶-12（caspase-12）mRNA及蛋白质表达。结果：①与N组相比，HH组、4-PBA组、TM组mPAP、右心室游离壁重量/左心室加心室间隔重量[RV/（LV+S）]、肺动脉管壁面积/管总面积（WA/TA）比值增加（P<0.0 1），肺动脉管腔面积/管总面积（LA/TA）比值减小（P<0.01），细胞凋亡指数降低（P <0.05或P<0.01）。ERS相关蛋白质及mRNA的表达量升高，各差异均有统计学意义。②与HH组相比，4-PB A组mPAP和[RV/（LV+S）]、WA/TA值减小（P<0.01），LA/TA值和细胞凋亡指数上升（P<0.05或P<0.01），ERS相关蛋白质和mRNA的表达量均下调（P<0.05或P<0.01）；③与HH组相比，TM组mPAP、[RV/（LV+S）]、WA/TA值升高（P<0.05或P<0.01）；肺动脉中膜层增厚，LA/TA值和细胞凋亡指数降低（P<0.01）。ERS相关蛋白质及mRNA的表达量均升高，除GRP78蛋白质表达量无明显变化外，其余各差异均有统计学意义。结论：低氧高二氧化碳诱导的肺动脉高压大鼠肺血管重塑可能与肺动脉平滑肌细胞增殖过度及凋亡过少有关；ERS相关因子（JNK、caspase-12和CHOP）参与低氧高二氧化碳性肺动脉高压的调控。

关键词: 内质网应激, 肺动脉高压, 低氧高二氧化碳, 大鼠

Abstract: Objective: To observe the pulmonary vascular remodeling in rats with pulmonary hypertension induced by hypoxia and hypercapnia, and to explore the role of endoplasmic reticulum stress in pulmonary hypertension.Methods: Forty SD rats were random-ly divided into four groups:normoxic control group (N), hypoxia hypercapnia group (HH), ERS inhibitor 4-phenylbutyric acid group (4-PBA), endoplasmic reticulum stress (ERS) pathway agonist tunicamycin group (TM), ten rats in each group.The mean pulmona-ry artery pressure (mPAP), mean carotid artery pressure (mCAP) and right ventricular hypertrophy index of rats in each group were measured.Pulmonary artery smooth muscle cells were identified by immunofluorescence α-smooth muscle actin (α-SMA).Morphologi-cal changes of lung tissue and pulmonary artery were observed by electron microscope.The apoptotic index of pulmonary artery smooth muscle cells in each group was detected by TUNEL.Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of glucose-regulated protein (GRP78), C/EBP homologous protein (CHOP), c-Jun N-terminal kinase (JNK) and cysteinyl aspartate specific proteinase-12 (caspase-12) mRNA and protein in each group.Results: ①Compared with the N group, the mPAP, the ratio of right ventricle weight to left ventricle plus ventricular septum weight[RV/(LV+S)]and the ratio of pulmonary artery wall area to total tube area (WA/TA) were increased (P<0.01), and the ratio of pulmonary artery luminal area to total tube area (LA/TA) were decreased (P<0.01), pulmonary artery smooth muscle cell apoptosis index were decreased (P<0.05 or P<0.01) in HH group, 4-PBA group and TM group.ERS related protein and mRNA expressions were increased, the differences were statistically significant.②Compared with the HH group, the mPAP, [RV/(LV+S)]and WA/TA of 4-PBA group were decreased (P <0.01), LA/TA and pulmonary artery smooth muscle cell apoptosis index were increased (P<0.01, P<0.05).The expressions of ERS related protein and mRNA were all decreased (P<0.05 or P<0.01).③Compared with the HH group, the mPAP, [RV/(LV+S)]and WA/TA of TM group were increased (P<0.05 or P<0.01), pulmonary artery middle layer thickened, LA/TA and pulmonary artery smooth muscle cell apoptotic index were decreased (P<0.01).ERS related protein and mRNA expressions were increased with statistical significance except GRP78 protein.Conclusion: Pulmonary vascular remodeling in rats with pulmonary hypertension induced by hypoxia and hypercapnia may be related to the excessive proliferation of pulmonary artery smooth muscle cells and too little apopto-sis;ERS related factors (JNK, caspase-12 and CHOP) are involved in the regulation of pulmonary hypertension induced by hypoxia hypercapnia.

Key words: endoplasmic reticulum stress, pulmonary artery hypertension, hypoxia hypercapnia, rat

张晶晶, CHEN Jun-hao, 赵美平, 武垣伶, 张聪聪, 应磊, 陈锡文, 王万铁. 内质网应激在大鼠低氧高二氧化碳性肺动脉高压中的作用[J]. 中国应用生理学杂志, 2018, 34(4): 327-333.

ZHANG Jing-jing, CHEN Jun-hao, ZHAO Mei-ping, WU Yuan-ling, ZHANG Cong-cong, YING Lei, CHEN Xi-wen, WANG Wan-tie. The role of endoplasmic reticulum stress in pulmonary hypertension in rat induced by chronic hypoxia and hypercapnia[J]. CJAP, 2018, 34(4): 327-333.

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内质网应激在大鼠低氧高二氧化碳性肺动脉高压中的作用

The role of endoplasmic reticulum stress in pulmonary hypertension in rat induced by chronic hypoxia and hypercapnia

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