基于自噬基因mRNA-microRNA研究白杨素抑制LPS所致RAW264.7细胞炎症的机制  

Mechanism of chrysin inhibiting RAW264.7 inflammation induced by LPS based on autophagy-related mRNA and microRNA

作　　者：胡子旋 蔡大可 李钰婷 黄雪君 甘海宁 黄丹娥 姚楠 陈玉兴 HU Zixuan;CAI Dake;LI Yuting;HUANG Xuejun;GAN Haining;HUANG Dane;YAO Nan;CHEN Yuxing(Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Guangdong Province Engineering Technology Research Institute of Chinese Medicine,Guangzhou 510095,China;Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine,Guangzhou 510095,China)

机构地区：[1]广州中医药大学第五临床医学院,广东广州510405 [2]广东省中医药工程技术研究院,广东广州510095 [3]广东省中医药研究开发重点实验室,广东广州510095

出　　处：《广东药科大学学报》2020年第6期820-826,共7页Journal of Guangdong Pharmaceutical University

基　　金：国家自然科学基金项目(81973508);广州市科技计划项目(2017A070701017);广州市科技计划项目(202002030215)。

摘　　要：目的探讨自噬在白杨素抑制脂多糖(LPS)诱导RAW264.7细胞炎症过程中的作用。方法使用CCK8检测不同浓度白杨素刺激下RAW264.7细胞的存活率。RAW264.7细胞(4×10^5/mL)培养24 h后,给予不同浓度的白杨素孵育24 h,组别分别为空白对照组,LPS模型组(0.1μg/mL),白杨素5μmol/L、0.5μmol/L、0.05μmol/L剂量组。RT-qPCR检测IL-1β、IL-6、COX-2、iNOS、IL-5、IL-10等炎症相关基因mRNA水平,beclin1、ULK1、LC3B、ATG5、ATG4B等自噬相关基因mRNA水平以及miR-26a-5p、miR-30e-5p、miR-124-3p、miR-155-5p、miR-199a-5p、miR-204-5p等自噬相关microRNA。结果白杨素能抑制促炎因子IL-1β、IL-6、COX-2、iNOS的表达及上调抗炎因子IL-5、IL-10的表达水平。同时,白杨素能够抑制自噬负性调控基因miR-26a-5p、miR-30e-5p、miR-124-3p、miR-155-5p、miR-199a-5p、miR-204-5p的表达以及上调自噬启动基因beclin1、ULK1、LC3B、ATG5、ATG4B的表达。结论白杨素能降低LPS所致的RAW264.7细胞炎症反应,其作用机制可能与白杨素调控自噬相关mRNA和microRNA有关。Objective To investigate the role of autophagy in the inhibition of lipopolysaccharide(LPS)-induced RAW264.7 cell inflammation by chrysin.Methods CCK8 was used to detect the survival rate of RAW264.7 cells stimulated by different concentrations of chrysin.RAW264.7 cells(4×10^5/mL)were incubated for 24 h and then treated with chrysin for another 24 h.The control group,model group(0.1μg/mL LPS),and LPS with different doses of chysin(at 5,0.5 and 0.05μmol/L)groups were set.RT-qPCR was used to detect the expression of pro-inflammation markers(IL-1β,IL-6,COX-2,iNOS),anti-inflammation markers(IL-5,IL-10),autophagy genes(beclin1,ULK1,LC3B,ATG5,ATG4B)and autophagy-related microRNA(miR-26a-5p,miR-30e-5p,miR-124-3p,miR-155-5p,miR-199a-5p,miR-204-5p).Results Chrysin inhibited the expression of pro-inflammatory genes IL-1β,IL-6,COX-2,iNOS and up-regulated the expression of anti-inflammatory genes IL-5,IL-10.Meantime,chrysin decreased the expression of miR-26a-5p,miR-30e-5p,miR-124-3p,miR-155-5p,miR-199a-5p,miR-204-5p and enhanced the expression of autophagy genes beclin1,ULK1,LC3B,ATG5 and ATG4B.Conclusion Chysin can reduce LPS-induced RAW264.7 cell inflammation,which may be related to regulation of autophagy and microRNA.

关 键 词：白杨素 炎症 自噬 microRNA RAW264.7细胞 

分 类 号：R285[医药卫生—中药学]