摘要
目的观察石斛酚(gigantol)对同型半胱氨酸(homocysteine,Hcy)诱导的人微血管内皮细胞(HMEC-1)NLRP3炎性体激活的影响,分析石斛酚对血管内皮细胞炎症反应的抑制作用及机制。方法采用形态学观察和MTT法检测细胞活性;采用ELISA法检测白细胞介素-1β(IL^-1β)浓度;采用Western blot法分析NLRP3、ASC、Caspase-1和TXNIP蛋白表达;采用流式细胞仪检测细胞活性氧(ROS)水平。结果Hcy(100μmol·L^-1)、石斛酚(1~10μmol·L^-1)对HMEC-1细胞活性未观察到明显影响;Hcy可明显增加细胞ROS水平,诱导TXNIP、NLRP3、ASC和Caspase-1蛋白表达,促进HMEC-1细胞IL^-1β分泌;石斛酚能明显逆转Hcy上调的HMEC-1细胞ROS和上清液IL^-1β水平,抑制Hcy诱导的TXNIP蛋白过表达和NLRP3炎性体激活。结论石斛酚能逆转Hcy激活的HMEC-1细胞NLRP3炎性体,抑制血管内皮细胞炎症反应,ROS/TXNIP通路可能参与石斛酚对Hcy诱导的NLRP3炎性体激活的抑制作用。
Aim To investigate the effects of gigantol on homocysteine(Hcy)induced NLRP3 inflammasomeactivation in human microvascular endothelial cells(HMEC-1)and the mechanisms responsible for its anti-inflammatory effects.Methods Cell viability was evaluated by MTT method and morphology observation.Proinflammatory cytokine IL^-1βwas detected by ELISA.The protein levels of NLRP3,ASC,Caspase-1,and TXNIP were determined by Western blot.The level of intracellular reactive oxygen species(ROS)was monitored by flow cytometry.Results Hcy(100μmol·L^-1)or gigantol(1~10μmol·L^-1)had no significant effect on HMEC-1 cell viability.Hcy significantly increased the level of intracellular ROS and IL^-1βin the cell culture supernatant,up-regulated the protein expression of NLRP3,ASC,Caspase-1 and TXNIP in HMEC-1 cells.Gigantol markedly decreased the level of intracellular ROS and IL^-1β,inhibited Hcy induced overexpression of TXNIP and NLRP3 inflammasome activation in HMEC-1 cells.Conclusions Gigantol can inhibit Hcy induced NLRP3 inflammasome activation in HMEC-1 cells,which may be through ROS/TXNIP pathway.
作者
王伟
陈亚军
耿明
徐忠东
戴敏
WANG Wei;CHEN Ya-jun;GENG Ming;XU Zhong-dong;DAI Min(School of Life Science, Hefei Normal University, Hefei 230601, China;Dept of Pharmacy, Anhui University of Traditional Chinese Medicine, Key Lab of Chinese Medicine Research and Development and Key Lab of Xin’an Medicine, Ministry of Education, Hefei 230031, China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2020年第8期1089-1094,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81773937)
安徽省高校自然科学研究重点项目(No KJ2020A0093)
安徽省高校优青重点项目(No gxyqZD2016235)
安徽省自然科学基金面上项目(No 2008085MH270)。
