五步蛇蛇毒对脂多糖诱导的RAW264.7巨噬细胞炎症反应的影响研究  

Effect of agkistrodon acutus venom on RAW264.7 macrophages inflammatory response induced by lipopolysaccharide

作　　者：曾晓艳[1] 林雅丽 刘应蛟 黄玉婷 龚力民[1] 刘塔斯[1] ZENG Xiao-yan;LIN Ya-li;LIU Ying-jiao;HUANG Yu-ting;GONG Li-min;LIU Ta-si(School of Pharmacy,Hunan University of Chinese Medicine,Changsha 410000;School of Pharmacy,Jiangxi University of Chinese Medicine,Nanchang 330004)

机构地区：[1]湖南中医药大学药学院,长沙410000 [2]江西中医药大学药学院,南昌330004

出　　处：《中南药学》2022年第8期1820-1824,共5页Central South Pharmacy

基　　金：2019年湖南省自然科学基金项目(No.2019JJ50435);2021年中央财政国家重点野生动植物保护资金项目(No.2130211)。

摘　　要：目的观察蛇毒对于脂多糖(LPS)诱导的RAW264.7巨噬细胞炎症的影响,并初步探索其作用机制。方法采用LPS诱导RAW264.7细胞构建立体外炎症模型,并通过ELISA法检测蛇毒25、50和100μg·mL^(-1)对TNF-α、IL-6和NO炎症因子分泌或生成的影响;Western blot法检测细胞中核转录因子(NF-κB P65)、核转录因子κB抑制剂激活酶(IκK)及核转录因子κB抑制剂(IκB)的蛋白表达水平。结果蛇毒质量浓度在0~200μg·mL^(-1)时,对RAW264.7细胞无毒性作用;质量浓度为25、50和100μg·mL^(-1)时对炎症模型分泌细胞因子TNF-α、IL-6和NO有明显的抑制作用,呈剂量依赖性;Western blot结果显示,蛇毒50μg·mL^(-1)和100μg·mL^(-1)能显著降低NF-κB p65及IκK蛋白表达水平,升高IκB表达水平。结论蛇毒对LPS诱导的RAW264.7巨噬细胞具有抗炎作用,其抗炎活性可能通过抑制TNF-α、IL-6和NO炎症因子分泌和生成实现,其作用机制可能与抑制NF-κB p65表达有关。Objective To observe the effect of agkistrodon acutus venom on lipopolysaccharide(LPS)induced RAW264.7 macrophages inflammation and determine its mechanism.Methods RAW264.7 cells were stimulated by LPS,and the model of inflammation in vitro was established.The effect of agkistrodon acutus venom(25,50 and 100μg·mL^(-1))on the inflammation model and on the secretion inflammatory factors TNF-α,IL-6 and NO was detected by ELISA.Meanwhile,the expression levels of nuclear transcription factor(NF-κB p65),nuclear factorκB inhibitor activator(IκK)and nuclear factorκB inhibitor(IκB)in each group were detected by Western blot.Results Agkistrodon acutus venom at 0-200μg·mL^(-1) had no effect on the RAW264.7 cells.Agkistrodon acutus venom at 25,50 and 100μg·mL^(-1) significantly inhibited the secretion of inflammatory factors TNF-α,IL-6 and NO dose-dependently(r=0.957,P<0.01).Western blot showed that the protein expression levels of NF-κB p65 and IκK in 50μg·mL^(-1) and 100μg·mL^(-1) venom were significantly decreased(P<0.001),while the expression level of IκB was significantly increased(P<0.01).Conclusion Agkistrodon acutus venom has anti-inflammatory effects on RAW264.7 macrophages induced by LPS,whose anti-inflammatory activity is by inhibiting the secretion and production of TNF-α,IL-6 and NO,and also the inhibition of NF-κB p65 expression.

关 键 词：蛇毒 脂多糖 RAW264.7细胞 细胞因子 核因子ΚB 

分 类 号：R284[医药卫生—中药学]