發(fā)布時(shí)間：2018-06-05 00:11

  本文選題：肌細(xì)胞 + 平滑肌　； 參考：《中國現(xiàn)代醫(yī)學(xué)雜志》2017年07期

【摘要】：目的探討缺氧條件下,細(xì)胞共培養(yǎng)體系中人肺動脈平滑肌細(xì)胞(HPASMCs)經(jīng)由Survivin信號通路調(diào)控人肺動脈內(nèi)皮細(xì)胞(HPAECs)的增殖。方法建立Transwell共培養(yǎng)體系將HPASMCs和HPAECs分別接種于下室和上室,根據(jù)HPASMCs是否行YM155(Survivin抑制劑)預(yù)處理進(jìn)行實(shí)驗(yàn)分組:常氧條件下HPASMCs與HAPECs共培養(yǎng)對照組(N組),常氧條件下YM155(終濃度100 nmol/l)預(yù)處理HPASMCs與HPAECs共培養(yǎng)組(NY組),缺氧條件下HPASMCs與HAPECs共培養(yǎng)組(H組)和缺氧條件下YM155預(yù)處理HPASMCs與HPAECs共培養(yǎng)組(HY組)。采用活細(xì)胞計(jì)數(shù)(CCK8)法檢測各組中HPASMCs與HPAECs的增殖活性(吸光度,A值),實(shí)時(shí)定量PCR檢測HPASMCs中Survivin m RNA的表達(dá),Western blot檢測HPASMCs中Survivin蛋白的表達(dá)。結(jié)果 N組HPASMCs與HPAECs增殖活性(0.561±0.007)、(0.619±0.013)與H組(0.777±0.030)、(0.875±0.021)比較,差異有統(tǒng)計(jì)學(xué)意義(q=16.615和21.333,P0.05)。HY組HPASMCs與HPAECs增殖活性為(0.661±0.027)、(0.723±0.025),與H組比較差異有統(tǒng)計(jì)學(xué)意義(q=8.923和12.667,P0.05)。共培養(yǎng)體系中N組HPASMCs未見m RNA和Survivin蛋白(0.017±0.001)表達(dá),H組中HPASMCs可見m RNA(4 506±849)和Survivin蛋白(0.932±0.018)表達(dá)。HY組HPASMCs m RNA和Survivin蛋白含量為(677±183)和(0.426±0.022),與H組比較差異有統(tǒng)計(jì)學(xué)意義(q=15.25和50.6,P0.05)。結(jié)論缺氧導(dǎo)致HPASMCs和HPAECs異常增殖,缺氧條件下共培養(yǎng)體系中HPASMCs經(jīng)由Survivin信號通路調(diào)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 鹵0.013) and that in group H (0.777 鹵0.030) was 0.875 鹵0.021). The proliferative activities of HPASMCs and HPAECs in group N and group H were 0.661 鹵0.027 and 0.723 鹵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 鹵0.001). In group H, m RNA(4 506 鹵849) and Survivin protein were 0.932 鹵0.018). The contents of HPASMCs m RNA and Survivin protein in group HY were 677 鹵183) and 0.426 鹵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.
【作者單位】： 空軍總醫(yī)院呼吸內(nèi)科;空軍總醫(yī)院腫瘤內(nèi)科;
【基金】：國家自然科學(xué)基金(No:81170045)
【分類號】：R544.1

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