绿原酸、咖啡酸、阿魏酸干预补体旁路激活致内皮细胞炎症反应的研究

发布时间：2018-03-11 04:16

本文选题：补体　切入点：补体旁路激活　出处：《贵州大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:研究绿原酸及其代谢产物咖啡酸和阿魏酸对补体旁路激活导致人微血管内皮细胞炎症反应中相关粘附分子表达的干预作用。方法:采用眼镜蛇毒因子(CVF)激活血清补体旁路,激活产物作用于人微血管内皮细胞,分别取多个时间点细胞培养上清,采用ELISA方法检测ICAM-1、IL-6、IL-8、t-PA、PAI-1的含量,MTT法筛选多个不同浓度的绿原酸、咖啡酸、阿魏酸对内皮细胞活力的影响,以确定合适的实验浓度,采用对细胞活力无影响的绿原酸、咖啡酸、阿魏酸浓度预处理内皮细胞,检测细胞培养上清中ICAM-1、IL-6、IL-8、t-PA、PAI-1的含量,对细胞活力有影响的浓度,进一步研究对内皮细胞的损伤情况。结果:补体旁路途径的激活物作用于微血管内皮细胞后,导致内皮细胞活化,引起细胞上清中ICAM-1、IL-6、IL-8、t-PA、PAI-1的浓度上调。0.1、0.5 mmol·L-1的绿原酸、咖啡酸、阿魏酸对细胞活力没有影响,1.0、3.0、5.0 mmol·L-1绿原酸、咖啡酸、阿魏酸组对细胞活力有明显的抑制作用。50、100、250μmol·L-1的绿原酸、咖啡酸、阿魏酸可下调ICAM-1、IL-6、IL-8的表达。而大于1.0 mmol·L-1绿原酸、咖啡酸、阿魏酸可导致细胞形态明显改变,细胞收缩,间隙变大,LDH释放在24 h时间点明显增加,各实验组的凋亡细胞比例明显升高。结论:50、100、250μmol·L-1的绿原酸、咖啡酸、阿魏酸对炎症反应有干预作用。而大于1.0 mmol·L-1的绿原酸、咖啡酸、阿魏酸可以造成内皮细胞损伤,引起内皮细胞凋亡。
[Abstract]:Objective: to study the effect of Lv Yuan acid and its metabolites caffeic acid and ferulic acid on the expression of associated adhesion molecules in inflammatory response of human microvascular endothelial cells induced by complement bypass activation. Huoxue Qing complement bypass, The supernatant of human microvascular endothelial cells (MECs) was cultured at different time points. The contents of IL-6, IL-8 and PAI-1 were detected by ELISA assay. The effects of different concentrations of Lv Yuan, caffeic acid and ferulic acid on the activity of endothelial cells were screened by MTT assay. In order to determine the appropriate experimental concentration, the endothelial cells were pretreated with the concentration of Lv Yuan, caffeic acid and ferulic acid, which had no effect on the cell viability. The concentration of ICAM-1, IL-6, IL-8 and PAI-1 in the supernatant of the cell culture was detected, and the concentration of the cell viability was determined. Results: after the activator of complement bypass pathway acted on microvascular endothelial cells, the endothelial cells were activated, and the concentration of ICAM-1, IL-6, IL-8 and PAPI-1 in the supernatant was up-regulated by the concentration of #number0# mmol 路L -1 Lv Yuan and caffeic acid in the supernatant. Ferulic acid had no effect on cell viability. The effects of caffeic acid, caffeic acid and ferulic acid on the cell viability were significant. 50 ~ 100 渭 mol 路L ~ (-1) Lv Yuan acid, caffeic acid, ferulic acid could down-regulate the expression of ICAM-1 IL-6IL-6IL-8, but more than 1.0 mmol 路L ~ (-1) Lv Yuan acid, caffeic acid, and ferulic acid could inhibit the expression of ICAM ~ (-1) IL-6IL-8, which was more than 1.0 mmol 路L ~ (-1) Lv Yuan acid, caffeic acid, and ferulic acid. Ferulic acid could obviously change the cell morphology, shrink the cells, increase the release of LDH at 24 h, and increase the proportion of apoptotic cells in each experimental group. Conclusion: the ratio of Lv Yuan and caffeic acid at #number0# 渭 mol 路L ~ (-1) is 100 渭 mol 路L ~ (-1). Ferulic acid could interfere with inflammatory reaction, while Lv Yuan, caffeic acid and ferulic acid more than 1.0 mmol 路L ~ (-1) could cause endothelial cell damage and induce endothelial cell apoptosis.
【学位授予单位】：贵州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R96

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