早发冠心病患者高密度脂蛋白抗炎作用的机制研究
发布时间：2018-10-25 10:59
【摘要】：目的:探讨早发冠心病患者高密度脂蛋白(High Density Lipoprotein,HDL)与健康受试者HDL相比对氧化型低密度脂蛋白(Oxidized Low-Density Lipoprotein,ox-LDL)诱导的人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVECs)炎症反应影响的区别及可能的机制,进一步明确早发冠心病的发病机制,为早发冠心病的临床防治提供新思路。方法:1.选取符合实验条件的健康受试者(男性55岁,女性65岁)及相应年龄的早发冠心病患者(男性55岁,女性65岁),采集血样,密度梯度离心法分离HDL;2.ox-LDL处理HUVECs不同时间,通过Elisa法检测细胞上清液中的炎症因子如高迁移率族蛋白B1(High Mobility Group Protein Box 1,HMGB-1)、肿瘤坏死因子-α(Tumor Necrosis Factor-α,TNF-α)、细胞间黏附分子-1(Intercellular Cell Adhesion Molecule-1,ICAM-1)的表达情况,通过Western Blot检测炎症通路分子HMGB-1及晚期糖基化终末产物受体(Receptor for Advanced Glycation End Product,RAGE)的表达情况,明确炎症反应最显著的时间点;3.不同浓度的HDL孵育HUVECs相应时间后,检测上述炎症分子及炎症通路分子的表达情况,明确HDL抗炎作用最显著的浓度;4.比较早发冠心病患者HDL和健康受试者HDL抗炎作用是否具有差异。结果:1.50mg/L ox-LDL可以诱导HUVECs产生炎症反应,随着时间的延长细胞上清液中HMGB-1、TNF-α及ICAM-1的水平升高(P0.05),在48小时达到高峰;细胞HMGB-1、RAGE蛋白表达量升高,在24小时达到高峰,48小时表达量降低,但仍较正常细胞表达量高,RAGE表达量一直处于升高状态,在48小时达到高峰(P0.05)。2.HDL对ox-LDL诱导的HUVECs的炎症反应具有抑制作用,细胞上清液中HMGB-1、TNF-α、ICAM-1的水平及细胞HMGB-1、RAGE蛋白表达量随着HDL浓度的增加而降低(P0.05),当HDL浓度达到200mg/L时上述炎症分子的表达量最低,说明此时HDL对炎症反应的抑制作用最显著(P0.05)。3.早发冠心病患者HDL组与健康受试者HDL组相比,细胞上清液中HMGB-1、TNF-α、ICAM-1的水平及细胞HMGB-1、RAGE蛋白表达量较高,但与模拟组相比上述分子水平较低,与空白组相比上述分子水平较高(P0.05)。结论:1.HDL可以通过抑制人脐静脉内皮细胞HMGB-1/RAGE炎症通路发挥抗炎作用。2.早发冠心病患者HDL与健康受试者HDL相比,其抗炎作用减弱,可能与其对HMGB-1/RAGE炎症通路的抑制作用减弱有关。提示HMGB-1/RAGE炎症通路可能作为早发冠心病防治的靶点。
[Abstract]:Objective: to investigate the effect of high density lipoprotein (High Density Lipoprotein,HDL) on oxidative low density lipoprotein (Oxidized Low-Density Lipoprotein,ox-LDL) -induced inflammation of human umbilical vein endothelial cells (Human Umbilical Vein Endothelial Cells,HUVECs) and its possible mechanism in patients with premature coronary heart disease (CHD) compared with healthy subjects (HDL). To further clarify the pathogenesis of early coronary heart disease and provide a new idea for clinical prevention and treatment of early onset coronary heart disease. Methods: 1. Blood samples were collected from healthy subjects (male 55 years old, female 65 years old) and premature coronary heart disease patients (55 years old for males and 65 years old for women). HDL;2.ox-LDL was separated by density gradient centrifugation to treat HUVECs for different time. The expression of inflammatory factors, such as high mobility group protein B1 (High Mobility Group Protein Box 1 (HMGB-1), tumor necrosis factor- 伪 (Tumor Necrosis Factor- 伪 (TNF- 伪) and intercellular adhesion molecule-1 (Intercellular Cell Adhesion Molecule-1,ICAM-1), was detected by Elisa assay. The expression of HMGB-1 and advanced glycosylation end product receptor (Receptor for Advanced Glycation End Product,RAGE) was detected by Western Blot to determine the most significant time point of inflammatory reaction. 3. After HUVECs was incubated with different concentrations of HDL, the expression of the above inflammatory molecules and inflammatory pathway molecules was detected, and the most significant anti-inflammatory effect of HDL was determined. 4. To compare the anti-inflammatory effects of HDL and HDL in patients with early coronary heart disease (CHD). Results: 1.50mg/L ox-LDL could induce the inflammatory reaction of HUVECs, and the levels of HMGB-1,TNF- 伪 and ICAM-1 in the supernatant of cells increased with time (P0.05) and reached the peak at 48 hours, the expression of HMGB-1,RAGE protein increased, reached the peak at 24 hours, and decreased at 48 hours. However, the expression of RAGE was still higher than that of normal cells, and the expression of RAGE reached its peak at 48 hours (P0.05). 2.HDL could inhibit the inflammatory response of HUVECs induced by ox-LDL. The levels of HMGB-1,TNF- 伪, ICAM-1 and the expression of HMGB-1,RAGE protein in the supernatant decreased with the increase of HDL concentration (P0.05). The lowest expression of the above inflammatory molecules was found when the HDL concentration reached 200mg/L, indicating that HDL had the most significant inhibitory effect on inflammatory response (P0.05). The levels of HMGB-1,TNF- 伪, ICAM-1 and the expression of HMGB-1,RAGE protein in the supernatant of the HDL group were higher than those in the control group, but lower than those in the simulated group and higher than those in the blank group (P0.05). Conclusion: 1.HDL can play an anti-inflammatory effect by inhibiting the HMGB-1/RAGE inflammatory pathway of human umbilical vein endothelial cells. 2. The anti-inflammatory effect of HDL in patients with premature coronary heart disease is weaker than that in healthy subjects compared with HDL, which may be related to the decrease of its inhibitory effect on the inflammatory pathway of HMGB-1/RAGE. The results suggest that the HMGB-1/RAGE inflammatory pathway may be a target for the prevention and treatment of early onset coronary heart disease.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R541.4
本文编号：2293513
[Abstract]:Objective: to investigate the effect of high density lipoprotein (High Density Lipoprotein,HDL) on oxidative low density lipoprotein (Oxidized Low-Density Lipoprotein,ox-LDL) -induced inflammation of human umbilical vein endothelial cells (Human Umbilical Vein Endothelial Cells,HUVECs) and its possible mechanism in patients with premature coronary heart disease (CHD) compared with healthy subjects (HDL). To further clarify the pathogenesis of early coronary heart disease and provide a new idea for clinical prevention and treatment of early onset coronary heart disease. Methods: 1. Blood samples were collected from healthy subjects (male 55 years old, female 65 years old) and premature coronary heart disease patients (55 years old for males and 65 years old for women). HDL;2.ox-LDL was separated by density gradient centrifugation to treat HUVECs for different time. The expression of inflammatory factors, such as high mobility group protein B1 (High Mobility Group Protein Box 1 (HMGB-1), tumor necrosis factor- 伪 (Tumor Necrosis Factor- 伪 (TNF- 伪) and intercellular adhesion molecule-1 (Intercellular Cell Adhesion Molecule-1,ICAM-1), was detected by Elisa assay. The expression of HMGB-1 and advanced glycosylation end product receptor (Receptor for Advanced Glycation End Product,RAGE) was detected by Western Blot to determine the most significant time point of inflammatory reaction. 3. After HUVECs was incubated with different concentrations of HDL, the expression of the above inflammatory molecules and inflammatory pathway molecules was detected, and the most significant anti-inflammatory effect of HDL was determined. 4. To compare the anti-inflammatory effects of HDL and HDL in patients with early coronary heart disease (CHD). Results: 1.50mg/L ox-LDL could induce the inflammatory reaction of HUVECs, and the levels of HMGB-1,TNF- 伪 and ICAM-1 in the supernatant of cells increased with time (P0.05) and reached the peak at 48 hours, the expression of HMGB-1,RAGE protein increased, reached the peak at 24 hours, and decreased at 48 hours. However, the expression of RAGE was still higher than that of normal cells, and the expression of RAGE reached its peak at 48 hours (P0.05). 2.HDL could inhibit the inflammatory response of HUVECs induced by ox-LDL. The levels of HMGB-1,TNF- 伪, ICAM-1 and the expression of HMGB-1,RAGE protein in the supernatant decreased with the increase of HDL concentration (P0.05). The lowest expression of the above inflammatory molecules was found when the HDL concentration reached 200mg/L, indicating that HDL had the most significant inhibitory effect on inflammatory response (P0.05). The levels of HMGB-1,TNF- 伪, ICAM-1 and the expression of HMGB-1,RAGE protein in the supernatant of the HDL group were higher than those in the control group, but lower than those in the simulated group and higher than those in the blank group (P0.05). Conclusion: 1.HDL can play an anti-inflammatory effect by inhibiting the HMGB-1/RAGE inflammatory pathway of human umbilical vein endothelial cells. 2. The anti-inflammatory effect of HDL in patients with premature coronary heart disease is weaker than that in healthy subjects compared with HDL, which may be related to the decrease of its inhibitory effect on the inflammatory pathway of HMGB-1/RAGE. The results suggest that the HMGB-1/RAGE inflammatory pathway may be a target for the prevention and treatment of early onset coronary heart disease.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R541.4
【参考文献】
相关期刊论文 前10条
1 李华明;欧志君;李艳;区景松;;高密度脂蛋白功能改变可能是动脉粥样硬化形成的危险因素[J];中国动脉硬化杂志;2015年12期
2 夏芳芳;栾天竹;梁兆光;姜超;杨建;;HMGB1与冠心病关系的研究进展[J];现代生物医学进展;2015年29期
3 潘兵;孔金阁;赵明明;郑乐民;;HDL通过ANXA1发挥对内皮细胞保护功能[J];中国病理生理杂志;2015年10期
4 杨惠月;边云飞;李小雷;尚冬升;;青年冠心病患者临床危险因素分析[J];山西医科大学学报;2015年05期
5 张舒媚;包金兰;孙润陆;黄灿霞;陈清群;周淑娴;张玉玲;;早发冠心病患者血清MCP-1与HDL2、HDL3水平及其相关性分析[J];中山大学学报(医学科学版);2015年03期
6 赵莉;姚树桐;王义围;秦树存;;高密度脂蛋白蛋白成分修饰对其抗动脉粥样硬化功能的影响[J];中国动脉硬化杂志;2014年10期
7 翟振丽;马维红;李全忠;申炜;;高密度脂蛋白功能差异与动脉粥样硬化的研究进展[J];实用医学杂志;2013年21期
8 于鹏;姜红;;高迁移率族蛋白1与心血管疾病关系概述[J];中国分子心脏病学杂志;2013年02期
9 韩菲菲;陈国千;;高迁移率族蛋白B1细胞外释放及信号转导机制[J];医学研究生学报;2013年03期
10 张婷;夏敏;;高迁移率族蛋白B1信号转导通路的研究进展[J];医学综述;2011年02期
相关博士学位论文 前1条
1 陈国良;早发冠心病患者高密度脂蛋白（HDL）蛋白质组学研究[D];中国协和医科大学;2010年
，本文编号：2293513
本文链接：https://www.wllwen.com/yixuelunwen/xxg/2293513.html
最近更新
教材专著
