人肺动脉平滑肌细胞对人肺动脉内皮细胞增殖的影响

发布时间：2018-06-05 00:11

  本文选题：肌细胞 + 平滑肌　； 参考：《中国现代医学杂志》2017年07期

【摘要】：目的探讨缺氧条件下,细胞共培养体系中人肺动脉平滑肌细胞(HPASMCs)经由Survivin信号通路调控人肺动脉内皮细胞(HPAECs)的增殖。方法建立Transwell共培养体系将HPASMCs和HPAECs分别接种于下室和上室,根据HPASMCs是否行YM155(Survivin抑制剂)预处理进行实验分组:常氧条件下HPASMCs与HAPECs共培养对照组(N组),常氧条件下YM155(终浓度100 nmol/l)预处理HPASMCs与HPAECs共培养组(NY组),缺氧条件下HPASMCs与HAPECs共培养组(H组)和缺氧条件下YM155预处理HPASMCs与HPAECs共培养组(HY组)。采用活细胞计数(CCK8)法检测各组中HPASMCs与HPAECs的增殖活性(吸光度,A值),实时定量PCR检测HPASMCs中Survivin m RNA的表达,Western blot检测HPASMCs中Survivin蛋白的表达。结果 N组HPASMCs与HPAECs增殖活性(0.561±0.007)、(0.619±0.013)与H组(0.777±0.030)、(0.875±0.021)比较,差异有统计学意义(q=16.615和21.333,P0.05)。HY组HPASMCs与HPAECs增殖活性为(0.661±0.027)、(0.723±0.025),与H组比较差异有统计学意义(q=8.923和12.667,P0.05)。共培养体系中N组HPASMCs未见m RNA和Survivin蛋白(0.017±0.001)表达,H组中HPASMCs可见m RNA(4 506±849)和Survivin蛋白(0.932±0.018)表达。HY组HPASMCs m RNA和Survivin蛋白含量为(677±183)和(0.426±0.022),与H组比较差异有统计学意义(q=15.25和50.6,P0.05)。结论缺氧导致HPASMCs和HPAECs异常增殖,缺氧条件下共培养体系中HPASMCs经由Survivin信号通路调控HPAECs的增殖。
[Abstract]:Objective to investigate the proliferation of human pulmonary artery endothelial cells (HPAECs) regulated by Survivin signal pathway in human pulmonary artery smooth muscle cells (HPA MCs) under hypoxia. Methods Transwell co-culture system was established to inoculate HPASMCs and HPAECs in lower chamber and superior chamber, respectively. According to whether HPASMCs was pretreated with YM155(Survivin inhibitor or not, the experimental group was divided into three groups: HPASMCs and HAPECs co-cultured control group under normoxic condition, YM155 (final concentration 100nmol / L) pretreatment HPASMCs and HPAECs co-culture group under normoxic condition, HPASMCs and HAPECs under hypoxia condition. HPASMCs was pretreated with YM155 and HPAECs co-cultured in HY group. The proliferative activity of HPASMCs and HPAECs (absorbance A value) was detected by living cell count (CCK8) method, the expression of Survivin m RNA in HPASMCs was detected by real-time quantitative PCR and the expression of Survivin protein in HPASMCs was detected by Western blot. Results the proliferative activity of HPASMCs and HPAECs in group N was 0.619 卤0.013) and that in group H (0.777 卤0.030) was 0.875 卤0.021). The proliferative activities of HPASMCs and HPAECs in group N and group H were 0.661 卤0.027 and 0.723 卤0.025, respectively, which were significantly higher than those in group H (P = 8.923 and P 0.050.05). In the co-culture system, the expression of m RNA and Survivin protein was not found in HPASMCs of group N (0.017 卤0.001). In group H, m RNA(4 506 卤849) and Survivin protein were 0.932 卤0.018). The contents of HPASMCs m RNA and Survivin protein in group HY were 677 卤183) and 0.426 卤0.022 (P < 0.05), which were significantly different from those in group H. Conclusion hypoxia leads to abnormal proliferation of HPASMCs and HPAECs, and HPASMCs regulates the proliferation of HPAECs through Survivin signaling pathway.
【作者单位】： 空军总医院呼吸内科;空军总医院肿瘤内科;
【基金】：国家自然科学基金(No:81170045)
【分类号】：R544.1

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