虫草素诱导巨噬细胞表达血红素氧合酶-1负向调控细胞因子分泌  

Cordycepin negatively modulates LPS-induced cytokine production by induction of Heme Oxygenase-1

作　　者：李旎[1] 刘玮[2] 赵兰华[2] 阳帆[3] 李冉辉[2] 

机构地区：[1]南华大学附属第二医院急诊内科,湖南省衡阳市421001 [2]南华大学医学院病原生物学研究所,421001 [3]湘南学院基础医学系,湖南省郴州市423043

出　　处：《实用医学杂志》2017年第8期1250-1254,共5页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:31500156)

摘　　要：目的:研究虫草素(cordycepin)负向调控脂多糖(LPS)诱导小鼠巨噬细胞分泌细胞因子的分子机制。方法:体外培养小鼠巨噬细胞RAW 264.7,用不同浓度的虫草素处理后,加入LPS刺激8 h。ELISA检测TNF-α、IL-6和IL-12的产生及细胞核内p65的含量;Western blot检测血红素氧合酶-1(HO-1)的表达以及IκB和p38的磷酸化;免疫荧光检测核转录因子Nrf2的核转位。结果:1~30μg/m L虫草素预处理RAW 264.7细胞后,可明显减少LPS处理后TNF-α、IL-6和IL-12的分泌,同时也能降低细胞核内NF-κB亚基p65含量以及IκB的磷酸化。此外,不同浓度的虫草素也能诱导RAW264.7细胞表达HO-1,并能诱导p38磷酸化以及Nrf2核转位。采用p38抑制剂SB203580预处理,或采用Nrf2 si RNA沉默其表达后,HO-1表达水平明显降低。而采用HO-1抑制剂处理后可逆转虫草素对细胞因子的抑制效应。结论:虫草素可能通过下调NF-κB的活性、激活p38和Nrf2诱导HO-1表达,最终发挥其抗炎效应。Objective To investigate the mocelualr mechanism of Cordycepin negative modulates LPS- induced cytokine production in murine macrophages. Methods The RAW264.7 murine macrophages were cultured in vitro and were pre-treated by different concentration of Cordycepin, and then stimulated by LPS for 8 h. Production of TNF-α, IL-6 and IL-12, and the content of p65 in the nuclear were detected by ELISA. Expression of heme oxygenase- 1 （HO- 1） and phosphorylation of IκB and p38 were measured by Western blot. Nuclear translocation of Nrf2 was detected by Immunofluorescence. Results 1 - 30 μg/mL of Cordycepin treatment significantly abrogated LPS-induced TNF-α, IL-6 and IL-12 production, p65 nuclear translocation and IκB phosphorylation. In addition, different concentration of Cordycepin could also induce RAW264.7 cells expression of HO- 1, phosphorylation of p38 and nuclear translocation of Nrf2. Application of p38 inhibitor and small interfering RNA-mediated knock-down of Nrf- 2 significantly inhibited surfactin- induced HO - 1 expression. Treatment with a selective inhibitor of HO- 1 reversed the Cordycepin mediated inhibition of pro-inflammatory cytokines. Conclusions Cordycepin induces anti- inflammatory effects by inhibition of NF-KB and activation of Nrf-2 and p38 mediated HO-1 induction.

关 键 词：虫草素 血红素氧合酶-1 小鼠巨噬细胞 

分 类 号：R285[医药卫生—中药学]