荆芥挥发油抗内毒素中毒小鼠NLRP3炎症小体通路的机制研究  

Study on mechanism of essential oils of Schizonepeta tenuifolia Briq. in endotoxin poisoning mice via NLRP3 inflammasome pathway

作　　者：吕红君 温桃群 罗杰 刘小波 杨菊 曾南[1] LYU Hong-jun;WEN Tao-qun;LUO Jie;LIU Xiao-bo;YANG-Ju;ZENG Nan(Key Lab of Systematic Research,Development and Utilization of Chinese Medicine Resources in Sichuan Province-Key Lab Breeding Base of Co-founded by Sichuan Province and MOST,Chengdu611137,China;State Key Lab of Biotherapy of Sichuan University,Chengdu610064,China)

机构地区：[1]成都中医药大学药学院中药材标准化教育部重点实验室中药资源系统研究与开发利用省部共建国家重点实验室培育基地,四川成都611137 [2]四川大学生物治疗国家重点实验室,四川成都610064

出　　处：《中国药理学通报》2019年第3期371-376,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81473399);成都市科技局科技惠民项目(No 2015-HM01-00187-SF)

摘　　要：目的以NLRP3炎症小体通路为切入点,探究荆芥挥发油抗炎效应的分子机制。方法荆芥挥发油低、高剂量(0.226、0.452 g·kg^(-1))连续灌胃给药5 d,末次给药后30 min,小鼠腹腔注射LPS (0.015 g·kg^(-1),10 mL·kg^(-1))构建内毒素中毒小鼠模型,造模后12 h取材,测定相关指标。Griess法测定肺组织NO含量;qPCR法检测肺组织中NLRP3、ASC、caspase-1、IL-1β、iNOS、p65 mRNA的表达;免疫组化法检测肺组织中P2X7R、Cathepsin B的表达;Western blot法测定肺组织中NLRP3、caspase-1(p20)、pro-IL-1β、COP1蛋白表达。结果 0.226 g·kg^(-1)荆芥挥发油明显降低模型小鼠肺组织Cathepsin B、NLRP3蛋白表达;0.452 g·kg^(-1)荆芥挥发油明显降低模型小鼠肺组织中NO水平,明显下调小鼠肺组织中NLRP3、iNOS、p65、IL-1βmRNA的表达及P2X7R蛋白表达;荆芥挥发油(0.226、0.452 g·kg^(-1))均能明显下调caspase-1(p20)蛋白水平,升高COP1蛋白水平。结论荆芥挥发油对内毒素中毒小鼠的保护作用与其抗炎效应密切相关,抗炎机制与抑制NLRP3炎症小体的激活有关。Aim To explore the mechanism of the anti-inflammatory effect of essential oils of Schizonepeta tenuifolia Briq.(EOST)by investigating the regulatory effect on NLRP3 inflammasome pathway.Methods EOST(0.226,0.452 g·kg^-1)were continuously administered for five days.Thirty minutes after the last administration,the mice were intraperitoneally injected with LPS(0.015 g·kg^-1,10 mL·kg^-1)to construct endotoxin poisoning,and relevant indicators for sample determination were taken 12 h after modeling.Determination of NO level in lung tissues by Griess method was performed.The expressions of NLRP3,ASC,caspase-1,IL-1β,iNOS and p65 mRNA in lung tissues were detected by qPCR.The expressions of P2X7R and Cathepsin B protein in lung tissues were assessed using immunohistochemistry.The NLRP3,caspase-1(p20),Pro-caspase-1,COP-1 protein ex pressions were detected by Western blot.Results 0.226 g·kg^-1 EOST could down-regulate the expressions of Cathepsin B and NLRP3 proteins in lung tissues of mice;0.452 g·kg^-1 EOST significantly reduced NO level,the expressions of NLRP3,iNOS,p65,IL-1βmRNA and P2X7R protein in lung tissues of mice;EOST(0.226,0.452 g·kg^-1)markedly reduced the caspase-1(p20)protein level and significantly increased COP1 protein level.Conclusions The protective effect of EOST on endotoxin poisoning mice is closely related to its anti-inflammatory effects,which involves the inhibition of activation of NLRP3 inflammasome.

关 键 词：荆芥挥发油 LPS 抗炎 NLRP3炎症小体 NO 分子机制 

分 类 号：R-332[医药卫生] R282.71