雌激素通过调节心肌细胞ⅠκBα发挥抗炎作用
发布时间：2018-07-19 20:00
【摘要】：本实验室之前的研究发现,缺氧/复氧(H/R)诱导原代培养心肌细胞核转录因子NF-κB p65入核,导致下游两种粘附分子ICAM-1,VCAM-1表达量增加,引起心肌细胞炎症反应。而雌激素(E2)通过抑制p65入核来降低ICAM-1,VCAM-1表达量,从而实现心肌保护作用。IκBα是NF-κB抑制蛋白IκB蛋白家族的主要成员,在调节NF-κB信号通路的激活和失活中起重要作用。因此,我们设计了这个实验用于检验雌激素对原代培养心肌细胞IκBα的影响。[实验目的]研究雌激素对缺氧/复氧后心肌细胞IκBα蛋白磷酸化、表达以及细胞内定位的影响,从而检验雌激素是否是通过调节细胞内IκBα来发挥抗炎作用。[实验方法]利用原代培养心肌细胞和已经检验建立成功的缺氧/复氧模型模拟心脏缺血再灌注,设立雌激素(5μM)孵育时间梯度(2h,4h,6h和8h),应用RT-PCR技术检测IκBαmRNA表达水平;western blot检测IκBα蛋白磷酸化(p-IκBα)水平、IκBα蛋白表达水平以及IκBα蛋白在胞浆和胞核中的量。[实验结果]与对照组比较,缺氧/复氧后心肌细胞内IκBα磷酸化水平显著增加,mRNA和蛋白表达水平以及胞核中IκBα蛋白水平显著降低。与缺氧/复氧组比较,缺氧/复氧后孵育雌激素,细胞内IκBα磷酸化水平显著降低,而IκBαmRNA和蛋白表达水平以及胞核中IκBα蛋白水平显著增加。[结论]雌激素通过抑制缺氧/复氧诱导的心肌细胞内IκBα蛋白磷酸化,促进IκBαmRNA和蛋白表达使细胞中IκBα蛋白保持一个相对稳定且较高的水平;同时雌激素还可以通过促进IκBα蛋白入核,使细胞核中活化的NF-κB失活,从而促进NF-κB由细胞核中输出,终止炎性因子转录反应,保护心肌细胞免受炎症反应的伤害,发挥起抗炎作用。
[Abstract]:Previous studies in our laboratory showed that hypoxia / reoxygenation (H / R) induced primary nuclear transcription factor NF- 魏 B p65 into the nucleus, resulting in an increase in the expression of ICAM-1 and VCAM-1, resulting in the inflammatory response of cardiomyocytes. Estrogen (E2) inhibits the expression of ICAM-1 and VCAM-1 by inhibiting p65 entry into the nucleus, and I 魏 B 伪 is a major member of the I 魏 B family of NF- 魏 B inhibitors, which plays an important role in regulating the activation and inactivation of NF- 魏 B signaling pathway. Therefore, we designed this experiment to test the effect of estrogen on I 魏 B 伪 in primary cultured cardiomyocytes. [objective] to study the effects of estrogen on the phosphorylation, expression and intracellular localization of I 魏 B 伪 protein in cardiomyocytes after hypoxia / reoxygenation, and to determine whether estrogen plays an anti-inflammatory role by regulating the intracellular I 魏 B 伪. [methods] using primary cultured cardiomyocytes and established a successful hypoxia / reoxygenation model to simulate myocardial ischemia reperfusion. Estradiol (5 渭 M) was incubated for 6 h and 8 h respectively. I 魏 B 伪 mRNA expression level was detected by RT-PCR and I 魏 B 伪 protein phosphorylation (p-I 魏 B 伪) level and I 魏 B 伪 protein in cytoplasm and nucleus were detected by western blot. [results] compared with the control group, I 魏 B 伪 phosphorylation in cardiomyocytes after hypoxia / reoxygenation significantly increased the expression of I 魏 B 伪 mRNA and protein, and significantly decreased the level of I 魏 B 伪 protein in the nucleus. Compared with hypoxia / reoxygenation group, the phosphorylation level of I 魏 B 伪 decreased significantly after hypoxia / reoxygenation, but I 魏 B 伪 mRNA and protein expression and I 魏 B 伪 protein in nucleus increased significantly. [conclusion] estrogen inhibits the phosphorylation of I 魏 B 伪 protein in cardiomyocytes induced by hypoxia / reoxygenation, and promotes the expression of I 魏 B 伪 mRNA and protein to maintain a relatively stable and high level of I 魏 B 伪 protein. At the same time, estrogen can inactivate the activated NF- 魏 B in the nucleus by promoting I 魏 B 伪 protein into the nucleus, thus promoting the output of NF- 魏 B from the nucleus, terminating the transcription of inflammatory factors, and protecting cardiomyocytes from inflammatory reaction. Play an anti-inflammatory effect.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
本文编号：2132918
[Abstract]:Previous studies in our laboratory showed that hypoxia / reoxygenation (H / R) induced primary nuclear transcription factor NF- 魏 B p65 into the nucleus, resulting in an increase in the expression of ICAM-1 and VCAM-1, resulting in the inflammatory response of cardiomyocytes. Estrogen (E2) inhibits the expression of ICAM-1 and VCAM-1 by inhibiting p65 entry into the nucleus, and I 魏 B 伪 is a major member of the I 魏 B family of NF- 魏 B inhibitors, which plays an important role in regulating the activation and inactivation of NF- 魏 B signaling pathway. Therefore, we designed this experiment to test the effect of estrogen on I 魏 B 伪 in primary cultured cardiomyocytes. [objective] to study the effects of estrogen on the phosphorylation, expression and intracellular localization of I 魏 B 伪 protein in cardiomyocytes after hypoxia / reoxygenation, and to determine whether estrogen plays an anti-inflammatory role by regulating the intracellular I 魏 B 伪. [methods] using primary cultured cardiomyocytes and established a successful hypoxia / reoxygenation model to simulate myocardial ischemia reperfusion. Estradiol (5 渭 M) was incubated for 6 h and 8 h respectively. I 魏 B 伪 mRNA expression level was detected by RT-PCR and I 魏 B 伪 protein phosphorylation (p-I 魏 B 伪) level and I 魏 B 伪 protein in cytoplasm and nucleus were detected by western blot. [results] compared with the control group, I 魏 B 伪 phosphorylation in cardiomyocytes after hypoxia / reoxygenation significantly increased the expression of I 魏 B 伪 mRNA and protein, and significantly decreased the level of I 魏 B 伪 protein in the nucleus. Compared with hypoxia / reoxygenation group, the phosphorylation level of I 魏 B 伪 decreased significantly after hypoxia / reoxygenation, but I 魏 B 伪 mRNA and protein expression and I 魏 B 伪 protein in nucleus increased significantly. [conclusion] estrogen inhibits the phosphorylation of I 魏 B 伪 protein in cardiomyocytes induced by hypoxia / reoxygenation, and promotes the expression of I 魏 B 伪 mRNA and protein to maintain a relatively stable and high level of I 魏 B 伪 protein. At the same time, estrogen can inactivate the activated NF- 魏 B in the nucleus by promoting I 魏 B 伪 protein into the nucleus, thus promoting the output of NF- 魏 B from the nucleus, terminating the transcription of inflammatory factors, and protecting cardiomyocytes from inflammatory reaction. Play an anti-inflammatory effect.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
【参考文献】
相关硕士学位论文 前2条
1 安尚玉;雌激素在缺氧/复氧心肌细胞中对NF-κB及粘附分子表达的影响[D];东北师范大学;2009年
2 钟世刚;雌激素对缺氧/复氧心肌核转录因子（NF-κB）及钙离子的影响研究[D];东北师范大学;2009年
，本文编号：2132918
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