17-β雌二醇通过雌激素受体α抑制肺动脉平滑肌细胞的增殖
发布时间：2018-08-16 18:16
【摘要】：目的:观察17-β雌二醇(E2)对低氧诱导的人肺动脉平滑肌细胞增殖(hPASMCs)的影响,并探讨雌激素受体(estrogen receptor,ER)在其中的介导作用。方法:1细胞培养:常氧组hPASMCs置于37℃、5%CO_2、95%氧气的培养箱中进行培养,低氧组hPASMCs置于37℃、5%CO_2、92%N2的三气培养箱中培养,每2d换液1次,并以0.25%胰蛋白酶进行消化传代,实验所用细胞为第5~8代,均处于对数生长期。实验前,细胞均在不含FBS的无酚红DMEM培养液中培养24h进行同步化。2细胞干预:应用第5~8代hPASMCs,分为常氧组、低氧组、低氧+E2(10-6mol/L)组、低氧+E2(10-7mol/L)组、低氧+E2(10-8mol/L)组、低氧+E2(10-9mol/L)组,行MTT实验测定不同干预条件下细胞增殖情况,筛选最适E2干预浓度。其次,将细胞分为常氧组、低氧组、低氧+E2(10-6mol/L)、低氧+E2(10-6mol/L)+MPP组、低氧+E2(10-6mol/L)+PHTPP组,行MTT实验测定各组细胞增殖情况,并采用RT-PCR法测定不同组hPASMCs中ERα、ERβ及PCNA的mRNA表达水平,采用Western blot法测定不同组hPASMCs中ERα、ERβ及PCNA的蛋白表达水平。结果:慢性低氧可显著刺激hPASMCs增殖(P0.01),应用E2干预后hPASMCs增殖减少(P0.01),且最佳抑制浓度为10-6mol/l。另外,慢性低氧诱导细胞增殖的同时伴随ERαmRNA及蛋白表达下降(P0.01),PCNAmRNA及蛋白表达升高(P0.01),而ERβmRNA及蛋白表达无明显变化(P0.05);应用E2干预后,抑制了细胞的增殖并逆转了ERα、PCNA指标的以上变化(P0.01),但对ERβ表达无明显影响(P0.05)。在低氧+E2基础上,应用MPP后可显著削弱E2的以上作用(P0.01),而应用ERβ抑制剂PHTPP干预则无明显影响(P0.05)。结论:1 E2能有效抑制hPASMCs的增殖。2 E2可能通过上调ERα的表达抑制hPASMCs的增殖,进而发挥其拮抗HPH的作用。
[Abstract]:AIM: To observe the effect of 17-beta estradiol (E2) on hypoxia-induced proliferation of human pulmonary artery smooth muscle cells (hPASMCs) and explore the mediating effect of estrogen receptor (ER). METHODS: 1 Cell culture: hPASMCs in normoxia group were cultured at 37 C, 5% CO_2, 95% oxygen, and hPASMCs in hypoxia group were cultured at 37 C and 5% CO_2. The cells were cultured in a three-gas incubator containing 92% N 2 and were subcultured with 0.25% trypsin every 2 days. The cells of the 5th to 8th passages were all in logarithmic growth phase. The cells were divided into normal oxygen group, hypoxia + E2 (10-6 mol/L), hypoxia + E2 (10-7 mol/L), hypoxia + E2 (10-8 mol/L), hypoxia + E2 (10-8 mol/L), hypoxia + E2 (10-8 mol/L), hypoxia + E2 (10-9 mol/L) group, and hypoxia + E2 (10-9 mol/L) group. MTT experiment was used to determine the cell proliferation under different intervention conditions, and the optimal concentration of E2 intervention was screen. Second, the cells were divided into normoxygen group, hypooxygen group, hypoxia + E2 (10-6 mol/L), hypoxia + E2 (10-6 mol/L), E2 (10-6 mol/+PHTPP group, MTT assay was used to determine the proliferation of hPASMCs, and RT-PCR was used to determine the mRNA expression levels of ERa, ERbeta and PCNA in hPASMCs of different groups. Western blot was used to determine the protein expression levels of ERa, ERbeta and PCNA in hPASMCs of different groups. The optimal inhibitory concentration was 10-6 mol/l. In addition, chronic hypoxia-induced cell proliferation was accompanied by decreased expression of ER-alpha mRNA and protein (P 0.01), increased expression of PCNA mRNA and protein (P 0.01), but no significant change of ER-beta mRNA and protein expression (P 0.05). On the basis of hypoxia + E2, MPP could significantly weaken the above effect of E2 (P 0.01), but PHTPP intervention had no significant effect (P 0.05). Conclusion: 1 E2 can effectively inhibit the proliferation of hPASMCs. 2 E2 may inhibit the proliferation of hPASMCs by up-regulating the expression of ERa, and then play its antagonism. Anti HPH effect.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R544.1
本文编号：2186784
[Abstract]:AIM: To observe the effect of 17-beta estradiol (E2) on hypoxia-induced proliferation of human pulmonary artery smooth muscle cells (hPASMCs) and explore the mediating effect of estrogen receptor (ER). METHODS: 1 Cell culture: hPASMCs in normoxia group were cultured at 37 C, 5% CO_2, 95% oxygen, and hPASMCs in hypoxia group were cultured at 37 C and 5% CO_2. The cells were cultured in a three-gas incubator containing 92% N 2 and were subcultured with 0.25% trypsin every 2 days. The cells of the 5th to 8th passages were all in logarithmic growth phase. The cells were divided into normal oxygen group, hypoxia + E2 (10-6 mol/L), hypoxia + E2 (10-7 mol/L), hypoxia + E2 (10-8 mol/L), hypoxia + E2 (10-8 mol/L), hypoxia + E2 (10-8 mol/L), hypoxia + E2 (10-9 mol/L) group, and hypoxia + E2 (10-9 mol/L) group. MTT experiment was used to determine the cell proliferation under different intervention conditions, and the optimal concentration of E2 intervention was screen. Second, the cells were divided into normoxygen group, hypooxygen group, hypoxia + E2 (10-6 mol/L), hypoxia + E2 (10-6 mol/L), E2 (10-6 mol/+PHTPP group, MTT assay was used to determine the proliferation of hPASMCs, and RT-PCR was used to determine the mRNA expression levels of ERa, ERbeta and PCNA in hPASMCs of different groups. Western blot was used to determine the protein expression levels of ERa, ERbeta and PCNA in hPASMCs of different groups. The optimal inhibitory concentration was 10-6 mol/l. In addition, chronic hypoxia-induced cell proliferation was accompanied by decreased expression of ER-alpha mRNA and protein (P 0.01), increased expression of PCNA mRNA and protein (P 0.01), but no significant change of ER-beta mRNA and protein expression (P 0.05). On the basis of hypoxia + E2, MPP could significantly weaken the above effect of E2 (P 0.01), but PHTPP intervention had no significant effect (P 0.05). Conclusion: 1 E2 can effectively inhibit the proliferation of hPASMCs. 2 E2 may inhibit the proliferation of hPASMCs by up-regulating the expression of ERa, and then play its antagonism. Anti HPH effect.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R544.1
【参考文献】
相关期刊论文 前2条
1 任亚浩;李岩溪;赵越;于飞;詹志鹏;袁媛;杨军;;白藜芦醇对C57BL/6J小鼠脂代谢的影响[J];卫生研究;2011年04期
2 许丽辉;何强;;Myocardin在大鼠血管平滑肌细胞表型转化中的作用机制[J];中国老年学杂志;2010年17期
，本文编号：2186784
本文链接：https://www.wllwen.com/yixuelunwen/xxg/2186784.html
最近更新
教材专著
