石斛酚对同型半胱氨酸诱导的人微血管内皮细胞NLRP3炎性体激活的影响  

Effects of gigantol on homocysteine induced NLRP3 inflammasome activation in human microvascular endothelial cells

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)

机构地区：[1]合肥师范学院生命科学学院,安徽合肥230601 [2]安徽中医药大学药学院,安徽省中药研究与开发重点实验室,新安医学教育部重点实验室,安徽合肥230031

出　　处：《中国药理学通报》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)。

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.

关 键 词：石斛酚 同型半胱氨酸 人微血管内皮细胞 NLRP3炎性体 活性氧物质 硫氧还蛋白互作蛋白 

分 类 号：R284.1[医药卫生—中药学] R322.12[医药卫生—中医学]