缺氧诱导心肌细胞凋亡中Bcl-2、Bax和NF-k B的表达及雌激素的作用

发布时间：2018-02-10 00:32

本文关键词： 缺氧心肌细胞凋亡 Bcl-2 Bax 核因子-κB 17β-雌二醇儿童青少年身体成分用力呼气流量用力肺活量生长突增　出处：《北京协和医学院》2012年硕士论文　论文类型：学位论文

【摘要】：[目的]探讨缺氧诱导心肌细胞凋亡中Bcl-2、Bax和NF-K B的表达,以及17β-雌二醇(E2)对缺氧心肌细胞的保护作用。 [方法]Sprague-Dawley大鼠乳鼠(出生1-2d)心肌细胞原代培养,待细胞生长良好时进行药物干预和缺氧刺激。将细胞分成四组,分别为C1(对照组)、C2(单纯加药组)、H1(单纯缺氧组)和H2(加药缺氧组)。即C2、H2经E2作用24h,之后H1、H2缺氧(5%CO2+95%N：)处理12h,再进行统一处理。采用MTT法检测缺氧对心肌细胞活力和抑制率的影响及E2对缺氧心肌细胞的保护作用。采用Flow cytometry方法检测心肌细胞的凋亡。采用Western Blot技术检测Bcl-2、Bax和NF-K B的表达变化。采用Real-Time PCR技术分析Bel-2mRNA和Bax mRNA水平的表达变化。 [结果]缺氧使心肌细胞活力下降,且对心肌细胞生长产生抑制,108、10-7mol/LE2可以提高心肌细胞活力,逆转缺氧对心肌细胞生长的抑制作用。缺氧加重心肌细胞凋亡,使心肌细胞Bax蛋白、Bax mRNA, NF-K B蛋白的表达升高,Bcl-2蛋白、Bcl-2mRNA的表达降低,而经10-7mol/L17β-雌二醇预处理的缺氧心肌细胞较单纯缺氧组心肌细胞Bax蛋白、Bax mRNA, NF-κB蛋白的表达降低,Bcl-2蛋白、Bcl-2mRNA表达升高。 [结论]缺氧对心肌细胞生长具有一定的抑制作用,还促进心肌细胞的凋亡,且使Bcl-2蛋白和mRNA表达降低、Bax蛋白和mRNA表达升高,NF-K B的蛋白表达升高。E2能够逆转缺氧对心肌细胞的抑制作用,这种保护作用可能通过降低NF-K B的表达进而上调Bcl-2mRNA的表达,下调Bax mRNA的表达来实现的。本文旨在研究儿童青少年去脂体重(fat free mass,FFM)、脂肪体重(fat mass, FM)和肺通气功能的关系,为儿童青少年相关疾病的预防、治疗以及健康管理提供参考数据。随机抽取黑龙江省10-18岁健康儿童青少年1174人(男性583人,女性591人),观察身高、体重和腰臀比(waist to hip ratio, WHR),同时采用身体成分测量仪和肺功能仪分别检测FFM、FM和肺通气功能,并采用独立样本t检验、偏相关和多元逐步回归分析FFM、FM和肺通气功能的关系。结果显示,受试者的性别、年龄均与去脂体重指数(FFM index,FFMI)呈正相关(P0.001)。男性年龄和脂肪体重指数(FM index,FMI)呈负相关(P0.001),女性年龄和FMI呈正相关(P0.001)。不同性别受试者FFMI与用力呼气肺活量(forced vital capacity, FVC)、用力呼气一秒量(forced expiratory volume in one second,FEV1)、最大呼气流量(peak expiratory flow, PEF)、用力呼出25%肺活量时呼气流量(forced expiratory flow at25%of forced vital capacity,FEF25%), FEF50%和最大呼气中段流量(maximum mid-expiratory flow, MMEF)都呈正相关(P0.05),与FEV1/FVC呈负相关(P0.01)。男性FFMI与FEF75%正相关(P0.05),女性FFMI与FEF75%不相关。男性FMI与FEV1、 FEV1/FVC、PEF、FEF25%、FEF50%、FEF75%和MMEF呈负相关(P0.05),而与FVC不相关。女性FMI与肺通气功能的指标均不相关。除男性FEV1/FVC和FEF75%外,不同性别受试者FFMI对肺通气功能指标的预测作用大于FMI。FFMI对男性肺通气功能预测作用大于女性。男性肺功能突增发生在12-15岁：FVC较前一个年龄段分别增加10.05%、20.41%、11.75%和9.00%,FEV1分别较前一年龄段增加11.38%、20.07%、12.01%和9.14%。女性肺功能突增发生在12、13和18岁：FVC较前一年龄段分别增加13.73%、9.07%和9.76%,FEV1较前一年龄段分别增加13.17%、11.12%和10.62%。在肺功能突增期,男女儿童青少年FFMI对肺功能的预测作用大于年龄的预测作用。综上,男女不同性别中,反映骨骼肌力的FFM均与肺通气功能呈正相关,对男性肺通气功能的预测作用大于女性。FM与男性肺通气功能呈负相关,而与女性肺通气功能不相关。在肺功能突增期,身高和FFM的快速增长可能是造成肺功能快速增长的主要原因。
[Abstract]:[Objective] to investigate the expression of Bcl-2, Bax and NF-K B in cardiomyocyte apoptosis induced by hypoxia, and the protective effect of 17 beta estradiol (E2) on the hypoxic myocardial cells.
Methods]Sprague-Dawley rats (born 1-2D) myocardial cells were cultured, when the cells grow well when drug intervention and hypoxia. The cells were divided into four groups, respectively C1 (control group), C2 (simple dosing group), H1 (hypoxia group) and H2 (chemical hypoxia group) at C2, H2 by E2 24h, H1, H2 (5%CO2+95%N:) 12h hypoxia treatment, and unified treatment. MTT method was used to detect the protective effect of hypoxia on myocardial cell activity and the inhibitory effect of E2 on hypoxic myocardial cells and myocardial cells. Apoptosis was detected by Flow cytometry method was used to detect the Bcl-2 Western. The technology of Blot, the expression of Bax and NF-K B. The expression of Bel-2mRNA Bax and mRNA Real-Time level by PCR technology.
[results] the myocardial hypoxia decreased the cell viability, and inhibit myocardial cell growth, 108,10-7mol/LE2 can improve the myocardial cell activity, inhibition of reverse hypoxia on the growth of cardiac myocytes. Hypoxia increased myocardial cell apoptosis, myocardial cell Bax protein, Bax mRNA, increased expression of NF-K B protein Bcl-2 protein, decreased the expression of Bcl-2mRNA however, hypoxic myocardial cells by 10-7mol/L17 beta estradiol pretreatment compared with hypoxia group, myocardial cell Bax protein, Bax mRNA, decreased expression of NF- kappa B protein Bcl-2 protein, the expression of Bcl-2mRNA increased.
[Conclusion] hypoxia has a certain inhibitory effect on myocardial cell growth, but also promote the apoptosis of myocardial cells, and Bcl-2 protein and mRNA expression decreased, increased expression of Bax protein and mRNA expression of NF-K B protein increased.E2 can reverse the inhibitory effect of hypoxia on myocardial cells, this protective effect may reduce the expression of NF-K expression by B then the upregulation of Bcl-2mRNA, downregulation of Bax mRNA.
The purpose of this paper is to study fat children and adolescents weight (fat free mass, FFM), fat mass (fat mass, FM) and the relationship between pulmonary function, prevention of related diseases in children and adolescents, and to provide reference data for health management. 10-18 years old in Heilongjiang province were randomly selected from healthy children and adolescents (583 males, 1174 females 591), observation height, weight and waist to hip ratio (waist to hip ratio, WHR), and the FFM were detected in body composition measurement and pulmonary function FM and pulmonary function, and the independent sample t test, partial correlation and multiple stepwise regression analysis, FFM, FM and pulmonary ventilation function the results showed that the subjects of sex, age and body fat index (FFM index, FFMI) were positively correlated (P0.001). The male age and fat mass index (FM, index, FMI) was negatively correlated (P0.001), female age and FMI positive correlation (P0.001) of different sex. The subjects of FFMI and forced expiratory vital capacity (forced vital capacity, FVC), forced expiratory volume in one second (forced expiratory volume in one second, FEV1), peak expiratory flow (peak expiratory, flow, PEF), forced expiratory expiratory flow at 25% of vital capacity (forced expiratory flow at25%of forced vital capacity, FEF25%, and FEF50%) maximal midexpiratory flow (maximum mid-expiratory, flow, MMEF) were positively correlated (P0.05), and negatively correlated with FEV1/FVC (P0.01). Male FFMI was positive correlation with FEF75% (P0.05), FFMI and FEF75% are not related to women. Male FMI and FEV1, FEV1/FVC, PEF, FEF25%, FEF50%, FEF75% and MMEF were negatively correlated (P0.05), but not correlated with FVC. The female FMI and pulmonary function indexes were not correlated. In addition to the male FEV1/FVC and FEF75%, different genders predict FFMI on pulmonary function index greater than FMI.FFMI of male pulmonary function pre The action is more than female. Male pulmonary function surge occurred in 12-15 years: FVC compared with the previous ages were increased by 10.05%, 20.41%, 11.75% and 9% FEV1 respectively, compared with the previous age increased by 11.38%, 20.07%, 12.01% and 9.14%. in female pulmonary function surge occurred in 12,13 and FVC than in the previous 18 years: age some were increased by 13.73%, 9.07% and 9.76% FEV1, compared with the previous ages were increased by 13.17%, 11.12% and 10.62%. in lung function spurt, predict FFMI male and female children and adolescents on lung function than predict age. In conclusion, different gender, reflect the muscle FFM and pulmonary function were positively correlated and the prediction effect on male lung function than female.FM was negatively correlated with male pulmonary function, but not associated with pulmonary function in lung function in women. The rapid growth spurt, height and FFM may be caused by the rapid increase in lung function The main reason for long.

【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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