大黄素通过调节Nrf2/HO-1和MAPKs抑制炎症和氧化应激机制研究  

Emodin inhibits inflammation and oxidative stress by regulating Nrf2/HO-1 and MAPKs

作　　者：张昊悦 赵蓓[1,2] 王业皇 章阳(指导)[1] ZHANG Hao-Yue;ZHAO Bei;WANG Ye-Huang;ZHANG Yang(Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Traditional Chinese Medicine,Nanjing 210000,China)

机构地区：[1]南京中医药大学附属南京中医院肛肠中心,南京210000 [2]上海中医药大学附属岳阳中西医结合医院,上海200437

出　　处：《中国免疫学杂志》2021年第9期1063-1068,共6页Chinese Journal of Immunology

基　　金：江苏省基础研究计划(自然科学基金)-青年基金项目(BK20180140);江苏省名老中医药专家传承工作建设项目(WYH-2016-JS);“十三五”南京市卫生青年人才培养工程(QRX17091)资助。

摘　　要：目的:研究大黄素对脂多糖(LPS)诱导炎症反应的抑制作用及分子机制。方法:CCK8检测大黄素对RAW264.7的最佳给药浓度;以LPS刺激RAW264.7细胞制备体外炎症模型,随机分为对照组、LPS组(LPS 1μg/ml)、大黄素低剂量组(LPS 1μg/ml+大黄素20μmol/L)和大黄素高剂量组(LPS 1μg/ml+大黄素40μmol/L),实时荧光定量PCR(RTPCR)检测细胞因子IL-6、iNOS、IL-1βmRNA的表达情况;Griess法检测各组NO含量;流式细胞术检测各组细胞内ROS的含量和细胞表面膜蛋白CD86阳性表达率;Western blot法检测大黄素干预RAW264.7后Nrf-2和HO-1的蛋白表达。同时检测大黄素对LPS诱导的RAW264.7细胞MAPK信号通路的影响。结果:CCK8结果显示,大黄素对RAW264.7细胞无明显毒副作用;大黄素呈剂量依赖地抑制由LPS诱导RAW264.7细胞中细胞因子IL-6、iNOS、IL-1βmRNA的表达量和NO含量;大黄素能明显抑制由LPS引起的RAW264.7细胞的ROS增多,且能够对LPS诱导RAW264.7细胞的膜蛋白CD86具有明显的抑制作用,呈浓度依赖性;大黄素对LPS诱导的Nrf-2/HO-1通路具有明显的抑制作用,并且对MAPK通路具有明显的抑制作用。结论:大黄素能调节Nrf-2/HO-1和MAPK信号通路,抑制由LPS诱导的RAW264.7细胞炎症和氧化应激。Objective:To investigate the inhibitory effect of Emodin on lipopolysaccharide(LPS)-induced inflammatory response and its molecular mechanism.Methods:The optimum concentration of emodin was evaluated by CCK-8 assay.The inflammatory model was established with LPS by stimulating RAW264.7 cells in vitro.Then all cells were divided into control group,model group(LPS 1μg/ml),emodin low dose group(LPS 1μg/ml+Emodin 20μmol/L),emodin high dose group(LPS 1μg/ml+emodin 40μmol/L).The mRNA levels of inflammatory cytokines IL-6,iNOS,IL-1βwere detected by qPCR.The effect of emodin on nitrite content in macrophage supernatant was determined by Griess reaction.The levels of ROS and CD86 were analyzed with flow cytometry;Western blot was used to detect Emodin after intervention with RAW264.7.Nrf-2 and HO-1 protein expression.The effects of Emodin on the MAPK signaling pathway of RAW264.7 cells induced by LPS were also detected.Results:CCK8 results showed that emodin had no significant toxic and side effects on RAW264.7 cells at 20 and 40μmol/L concentration;Emodin significantly inhibited the expression of IL-6,iNOS,and IL-1βmRNA induced by LPS in a dose-dependent manner.Compared with LPS group,the Emodin group not only significantly decreased the levels of NO,ROS,CD86;Emodin on LPS-induced Nrf-2/HO-1 pathway had obvious inhibitory effect,and MAPK pathway had obvious inhibitory effect.Conclusion:Emodin can regulate Nrf-2/HO-1 and MAPK signaling pathways in order to inhibit LPS-induced RAW264.7 cell inflammation and oxidative stress.

关 键 词：大黄素 炎性肠病 氧化应激 Nrf-2/HO-1 MAPK 

分 类 号：R28[医药卫生—中药学]