新型全蝎寡肽在体外抗苯并芘毒性的功能及其机制的评价  

Evaluation of the function and mechanism of novel whole scorpion oligopeptides against benzopyrene toxicity in vitro

作　　者：郑曼 袁燕 张风雷 马凯 李叶婷 顾磊 刘杰 王殿云 ZHENG Man;YUAN Yan;ZHANG Fenglei;MA Kai;LI Yeting;GU Lei;LIU Jie;WANG Dianyun(Department of Traditional Chinese Medicine,Dongying People's Hospital,Dongying 257091,China;Institute of Allergy and Immunology,Shenzhen University,Shenzhen 518060,China)

机构地区：[1]东营市人民医院,山东东营257091 [2]深圳大学过敏反应与免疫学研究所,广东深圳518060

出　　处：《广东药科大学学报》2023年第6期112-119,共8页Journal of Guangdong Pharmaceutical University

基　　金：山东省中医药科技项目(2021Q039)。

摘　　要：目的分离具有抗动脉粥样硬化(atherosclerosis,AS)活性的全蝎活性肽(SOPs)并研究其潜在分子机制。方法基于苯并芘(BaP)暴露的人脐静脉血管内皮细胞(HUVEC)模型分离、鉴定具有抗AS活性的SOPs;MTT法检测SOPs保护活性及SOP-1细胞毒性;ELISA法检测细胞炎性因子(IL-1β、IL-6、IL-8、PGE2及TNF-α)、细胞内ROS及ICAM-1、VCAM-1含量变化;Western blotting检测Pink1/Parkin和NLRP3炎症小体信号通路及COX-2蛋白的表达。结果分离、鉴定4条寡肽(SOP-1~4);其中,SOP-1能够显著提高HUVEC细胞活力,降低BaP诱导的细胞炎性因子(IL-1β、IL-6、IL-8、PGE2及TNF-α)及COX-2的表达;降低BaP诱导的HUVEC细胞内ROS含量;同时,SOP-1能够显著抑制BaP暴露诱导的HUVEC细胞ICAM-1过表达。SOP-1能抑制BaP诱导的HUVEC细胞Pink1/Parkin及NLRP3炎症小体信号通路的激活。结论SOP-1通过Pink1/Parkin和NLRP3炎症小体信号通路抑制细胞炎症因子和ROS的过量产生,从而抑制BaP的毒性作用,发挥抗AS活性。Objective To isolate scorpion bioactive peptides(SOPs)with anti-atherosclerosis(AS)activity and study their potential molecular mechanism.Methods Isolation and identification of SOPs with anti-AS activity based on a human umbilical vein endothelial cell(HUVEC)model exposed to benzopyrene(BaP).MTT assay was used to detect the protective activity of SOPs and the cytotoxicity of SOP-1 on cells.ELISA was used to measure inflammatory cytokines(IL-1β,IL-6,IL-8,PGE2,and TNF-α),and the levels of intracellular ROS,ICAM-1 and VCAM-1.Western blotting was used to detect the expression of Pink1/Parkin and NLRP3 inflammasome pathways and COX-2 proteins.Results Four oligopeptides(SOP-1-4)were isolated and identified.Among them,SOP-1 significantly increased HUVEC cell viability and reduced the expression of cytokines IL-1β,IL-6,IL-8,PGE2 and TNF-α,and COX-2 induced by BaP.SOP-1 reduced the intracellular ROS content and ICAM-1 overexpression in HUVEC cells exposed to BaP.SOP-1 inhibited the activation of Pink1/Parkin and NLRP3 inflammasome pathways in HUVEC cells exposed to BaP.Conclusion SOP-1 inhibits the excessive production of cytokines and ROS through the Pink1/Parkin and NLRP3 inflammasome pathways,thereby inhibiting the toxic effect of BaP and exerting anti-AS activity.

关 键 词：全蝎 寡肽 动脉粥样硬化 血管内皮细胞 Pink1/Parkin 

分 类 号：R965[医药卫生—药理学]