早发冠心病患者高密度脂蛋白抗HUVEC凋亡的作用机制

发布时间：2018-03-03 04:19

本文选题：早发冠心病　切入点：高密度脂蛋白　出处：《中国动脉硬化杂志》2017年07期 　论文类型：期刊论文

【摘要】：目的通过细胞实验探究早发冠心病(PCHD)患者高密度脂蛋白(HDLPCHD)与健康人群HDL(HDLhealth)抗人脐静脉内皮细胞(HUVEC)凋亡作用是否有区别及其可能机制。方法采集PCHD患者和与之相匹配的健康人血样,并提取HDL;不同浓度氧化型低密度脂蛋白(ox-LDL)处理HUVEC 24 h,MTT检测细胞存活率,明确ox-LDL引起HUVEC凋亡的合适浓度;不同浓度、不同时间的HDLhealth预处理HUVEC后,再用合适浓度的oxLDL处理HUVEC 24 h,MTT检测细胞存活率,明确HDLhealth预处理HUVEC的最适浓度与最适作用时间;用最适浓度HDLhealth、HDLPCHD对HUVEC进行最适作用时间的预处理,再用合适浓度的ox-LDL处理HUVEC 24 h,MTT检测细胞存活率。Annexin V-FITC/PI凋亡检测试剂盒进行流式细胞检测,Western blot检测Caspase 3、Caspase 9的蛋白表达,用试剂盒测定活性氧(ROS)活性,使用Lipoprint脂蛋白分析仪分析HDLhealth和HDLPCHD亚组分(HDL1-HDL10)分布情况。结果 100 mg/L ox-LDL处理HUVEC 24 h后细胞存活率为60.34%,较空白处理组明显降低(P0.05);200 mg/L HDLhealth预处理HUVEC 18 h后,细胞存活率为82.01%,可以明显减弱ox-LDL对细胞的损伤;200 mg/L HDLhealth显著抑制100 mg/L ox-LDL诱导的HUVEC凋亡及Caspase 3、Caspase 9的蛋白表达和ROS产生;200 mg/L HDLPCHD预处理HUVEC 18 h后,细胞存活率为65.5%,其作用较HDLhealth减弱;200 mg/L HDLPCHD可抑制100 mg/L ox-LDL诱导的HUVEC凋亡及Caspase 3、Caspase 9的蛋白表达和ROS产生,但作用较HDLhealth减弱;HDLPCHD亚组分大颗粒(HDL1-HDL3)含量较HDLhealth低(28.5%±5.7%比46.8%±15.2%),而小颗粒(HDL8-HDL10)含量较HDLhealth高(21.4%±7.8%比10.9%±5.4%)。结论 HDLPCHD与HDLhealth比较,可能由于其亚组分大颗粒(HDL1-HDL3)含量较HDLhealth低,而小颗粒(HDL8-HDL10)含量较HDLhealth高,导致其抗氧化功能减弱,抑制ox-LDL诱导内皮细胞凋亡的功能亦减弱,从而减弱或丧失抗动脉粥样硬化的作用。
[Abstract]:Objective to investigate the difference and possible mechanism of anti-apoptosis effect of high density lipoprotein (HDLPC) in patients with premature coronary heart disease (CHD) and healthy controls. Methods Blood samples from PCHD patients and matched healthy subjects were collected. Different concentrations of oxidized low density lipoprotein (ox-LDL) were used to detect the survival rate of HUVEC for 24 h, to determine the appropriate concentration of HUVEC apoptosis induced by ox-LDL, and to pretreat HUVEC with different concentration and time. The cell survival rate was determined by 24 h HUVEC treatment with appropriate concentration of oxLDL, and the optimal concentration and optimal time of HUVEC pretreatment with HDLhealth were determined, and the optimal time of HUVEC pretreatment with the optimal concentration of HDLhealth and HDLPCHD was determined. The cell survival rate. Annexin V-FITC / Pi apoptosis assay kit was used to detect the protein expression of Caspase 3 caspase 9 by flow cytometry, and the activity of reactive oxygen species (Ros) was measured by using the kit. Lipoprint lipoprotein analyzer was used to analyze the distribution of HDLhealth and HDLPCHD subfractions HDL1-HDL10. Results the cell survival rate of HUVEC treated with 100 mg/L ox-LDL for 24 h was 60.34, which was significantly lower than that of control group after HUVEC 18 h pretreated with P0.05 or 200 mg/L HDLhealth. The cell survival rate was 82.01, which could significantly attenuate the damage induced by ox-LDL and the apoptosis of HUVEC induced by 100 mg/L ox-LDL and the expression of Caspase _ 3 Caspase-9 protein and the production of Caspase _ (2) mg/L HDLPCHD pretreated with HUVEC for 18 h. The cell survival rate was 65. 5%, and its effect was lower than that of HDLhealth, which could inhibit HUVEC apoptosis induced by 100 mg/L ox-LDL and the expression of Caspase 3 Caspase 9 protein and the production of ROS. But the content of HDL1-HDL3) was lower than that of HDLhealth (28.5% 卤5.7% vs 46.8% 卤15.2g), while the content of HDL8-HDL10 was higher than that of HDLhealth (21.4% 卤7.8% vs 10.9% 卤5.4U). Conclusion compared with HDLhealth, the content of HDL1-HDL3 in the subcomponent of HDLPCHD is lower than that of HDLhealth, while the content of HDL8-HDL10 in small particles is higher than that of HDLhealth. The function of inhibiting the apoptosis of endothelial cells induced by ox-LDL was also weakened, which weakened or lost the effect of anti-atherosclerosis.
【作者单位】：山西医科大学;山西医科大学第二医院心内科心血管疾病诊治及临床药理山西省重点实验室;山西医科大学第一医院心内科;
【分类号】：R541.4

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