野漆树苷对LPS诱导的RAW264.7细胞的抗炎作用及机制探究

摘要:

目的:探究野漆树苷(Rhoifolin,RHO)对细菌脂多糖(lipopolysaccharide,LPS)诱导的小鼠单核巨噬细胞RAW264.7细胞炎症模型释放炎性因子的影响,并探讨其作用机制.方法:用脂多糖(0.5μg·mL-1)刺激体外生长良好的RAW264.7细胞24h建立体外细胞炎症模型,以MTT法测定不同浓度RHO对RAW264.7细胞的毒性作用,用Griess试剂法检测一氧化氮(Nitric Oxide NO)的含量,RT-PCR法检测细胞中肿瘤坏死因子α(Tumor Necrosis Factor,TNF-α),白介素1β(Interleukin-1β,IL-1β),白介素6(Interleukin,IL-6)的含量,采用Western Blot法检测MAPK信号通路中相关蛋白的含量.结果:与空白对照组相比,在LPS诱导下,RAW264.7细胞分泌致炎因子NO,TNF-α,IL-1β,IL-6(P<0.01);与模型组相比,25~100μmol·L-1的RHO可明显下调LPS诱导的RAW264.7细胞释放炎症因子NO,TNF-α,IL-1β和IL-6(P<0.05,P<0.01),并呈现良好的剂量依赖关系;野漆树苷还不同程度的抑制了Erk和JNK激酶的磷酸化.结论:RHO可以抑制LPS所致的RAW264.7细胞炎症反应,减少炎症因子NO分泌,抑制TNF-α,IL-1β和IL-6mRNA的合成,抑制JNK/SAPK及Erk信号通路可能是其抗炎作用机制之一.

This study performed to investigate the effect of Rhoifolin(RHO)on the secretion of inflammatory factors in monocyte-macrophage RAW264.7 cells induced by lipopolysaccharide(LPS),and to explore its mechanism.RAW264.7 cells were cultured with lipopolysaccharide(0.5μg·ml-1)for 24 hto establish the model of in vitro cell inflammation.MTT assay was used to determine the toxic effects of different concentrations of RHO on RAW264.7 cells.The levels of nitric oxide(NO)was determined by Griess reagent;the level of TNF-α,interleukin-1β(IL-1β)and interleukin-6(IL-6)in the cells were measured by RT-PCR;Western Blot was used to detect the content of related proteins in MAPK signal pathway.Data showed that compared with the control group,LPS could induce the secretion of inflammatory factors NO,TNF-α,IL-1βand IL-6(P<0.01)in RAW264.7 cells.Compared with the model group,25~100μmol·L1 RHO significantly down-regulated LPS-induced release of inflammatory cytokines NO,TNF-α,IL-1βand IL-6(P<0.05,P<0.01)and showed well dose-dependent relationship.In conclution,Rhoifolin also inhibited the phosphorylation of Erk and JNK kinase to some extent,RHO can inhibit the inflammatory response of RAW264.7 cells induced by LPS,and its anti-inflammatory mechanism may be related to the decrease of NO secretion,inhibition the level of TNF-α,IL-1βand IL-6 mRNA,and inhibit JNK/SAPK and Erk signaling pathways.

作者:

宋小欣 韩凌 李宇邦 陈家兰 林励 李海波 肖凤霞

机构地区:

广州中医药大学中药学院 广东省中医院 化州市正源堂化橘红实业有限公司

出处:

《betway官方app 学报：自然科学版》 CAS 北大核心 2018年第2期84-88,94,共6页

基金国家发展改革新兴产业重大工程包&国家中医药管理局中药标准化项目(ZYBZH-Y-GD-13)

关键词:

野漆树苷 脂多糖 肿瘤坏死因子α 白介素1Β 白介素6

Rhoifolin lipopolysaccharide tumor necrosis factorα interleukin 1β interleukin 6

分类号:

R285.5 [医药卫生—中药学]

野漆树苷对LPS诱导的RAW264.7细胞的抗炎作用及机制探究.pdf