载脂蛋白E对低氧诱导小鼠肺动脉平滑肌细胞增殖的影响及其机制*

doi:10.12047/j.cjap.5838.2019.090

摘要/Abstract

摘要： 目的:探讨外源性载脂蛋白E(apoE)对低氧诱导小鼠肺动脉平滑肌细胞(PASMCs)增殖的影响及其机制。方法:采用组织块贴壁法原代培养小鼠PASMCs,取对数生长期PASMCs,分常氧组、常氧+apoE组、低氧组和低氧+apoE组,常氧组培养条件为:21% O₂、5% CO₂,低氧组培养条件为:1% O₂、5% CO₂,外源性加apoE使终浓度为10 μg/ml,培养时间为48 h,重复三次。EdU掺入法检测细胞增殖情况,Western blot法检测apoE、增殖细胞核抗原(PCNA)、蛋白激酶C(PKC)和磷酸化蛋白激酶C(p-PKC)蛋白的表达。结果:与常氧组比较,低氧组PASMCs增殖率提高64.7%,PCNA蛋白和p-PKC蛋白表达分别上调69.0%和120.0%,而apoE蛋白表达下调51.0%(P均＜0.05);与低氧组比较,低氧+apoE组PASMCs增殖率降低19.6%,PCNA蛋白和p-PKC蛋白表达分别下调19.8%和103.2%(P均＜0.05);各组间PKC蛋白表达无显著性差异,常氧组p-PKC蛋白表达与常氧+apoE组的相比也无显著性差异(P均＞0.05)。结论:apoE能抑制低氧诱导小鼠PASMCs增殖,其机制可能与阻碍PKC途径有关。

关键词: 低氧, 肺动脉平滑肌细胞, 载脂蛋白E, 增殖

Abstract: Objective: To investigate the effects of apolipoprotein E (apoE) on the proliferation of pulmonary arterial smooth muscle cells (PASMCs) induced by hypoxia. Methods: Primary culture of mouse PASMCs was prepared from male C57BL/6 mouse pulmonary artery by the method of tissue block anchorage. PASMCs were divided into four groups: normoxia group, normoxia with apoE administration group, hypoxia group and hypoxia with apoE administration group. The proliferation of PASMCs was observed by EdU incorporation. The protein levels of apoE, proliferating cell nuclear antigen (PCNA), protein kinase C (PKC) and phosphorylated protein kinase C (p-PKC) were analyzed by Western blot.Results: The percentage of PASMCs proliferation of hypoxia group was significantly higher than that of normoxia group by 64.7% (P＜0.05), and the protein expression levels of PCNA and p-PKC of hypoxia group were up-regulated than those of normoxia group by 69.0% and 120.0%, while the protein expression of apoE was down-regulated by 51.0% (P＜0.05), respectively. The percentage of PASMCs proliferation of hypoxia with apoE administration group was significantly lower than that of hypoxia group by 19.6% (P＜0.05), and the protein expression levels of PCNA and p-PKC of hypoxia with apoE administration group were down-regulated than those of hypoxia group by 19.8% and 103.2% (P＜0.05), respectively. There was no significant difference among each group in the protein expression of PKC, nor do there any significant difference between normoxia group and hypoxia group in the protein expression of p-PKC (P＞0.05). Conclusion: ApoE can inhibit the proliferation of PASMCs induced by hypoxia, and the mechanism of its effect may be attributed to blocking PKC pathway.

Key words: hypoxia, pulmonary arterial smooth muscle cell, apolipoprotein E, proliferation

郝家乐, 徐立聪, 黄天鹏, 余艳, 王瑶尧, 范小芳, 龚永生, 毛孙忠. 载脂蛋白E对低氧诱导小鼠肺动脉平滑肌细胞增殖的影响及其机制^*[J]. 中国应用生理学杂志, 2019, 35(5): 414-417.

HAO Jia-le, XU Li-cong, HUANG Tian-peng, YU Yan, WANG Yao-yao, FAN Xiao-fang, GONG Yong-sheng, MAO Sun-zhong. Effects of apolipoprotein E on proliferation of mouse pulmonary arterial smooth muscle cells induced by hypoxia[J]. CJAP, 2019, 35(5): 414-417.

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载脂蛋白E对低氧诱导小鼠肺动脉平滑肌细胞增殖的影响及其机制^*

Effects of apolipoprotein E on proliferation of mouse pulmonary arterial smooth muscle cells induced by hypoxia

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