摘要
目的观察姜黄素对经LPS和IFN-γ诱导的RAW264.7巨噬细胞(M1)脂质转运及炎症的影响。方法LPS和IFN-γ诱导RAW264.7巨噬细胞12 h成M1表型。M1巨噬细胞分别进行不同处理:①不同浓度姜黄素(0,6.25,12.5μmol/L)对M1巨噬细胞进行干预;②不同浓度姜黄素(0,6.25,12.5μmol/L)和30 mg/L oxLDL共同干预M1巨噬细胞;③不同浓度姜黄素(0,6.25,12.5μmol/L)和NBD胆固醇与Apo-AI共同干预M1巨噬细胞;④20μmol/L GW9662与12.5μmol/L姜黄素共同干预经oxLDL或NBD胆固醇与Apo-AI刺激的M1巨噬细胞。分别采用油红O染色、BODIPY荧光染色方法检测M1巨噬细胞胆固醇流入/流出,用RT-PCR、Western Blot检测IL-1β、IL-6、ABCA1、CD36、PPARγmRNA及蛋白表达,用ELISA检测TNF-α、MMP-9炎症因子表达。结果姜黄素能减轻M1型巨噬细胞及oxLDL刺激的M1型巨噬细胞的炎症反应(P<0.05);姜黄素能增加oxLDL刺激的M1型巨噬细胞胆固醇流入及NBD胆固醇与Apo-AI介导的M1型巨噬细胞胆固醇流出(P<0.05);姜黄素能上调oxLDL刺激的M1型巨噬细胞CD36和PPARγ表达,姜黄素能上调NBD胆固醇与Apo-AI介导的M1型巨噬细胞ABCA1表达,减少TNF-α和MMP-9的表达(均P<0.05)。结论姜黄素可通过活化PPARγ通路增强M1型巨噬细胞处理有害脂质的能力,促进脂质的加工、清理和去除,维持胆固醇稳态,并减轻其炎症反应。
Objective To observe the effect of curcumin on cholesterol transport homeostasis and inflammation in RAW264.7 macrophages(M1)induced by LPS and IFN-γ.Methods RAW264.7 macrophages(M0)were treated with LPS plus IFN-γfor 12 h to deve-lop a M1 subtype.①M1 subtype macrophages were treated with different concentrations of curcumin(0,6.25,12.5μmol/L).②M1 subtype macrophages were treated with different concentrations of curcumin(0,6.25,12.5μmol/L)and oxLDL.③M1 subtype macrophages were treated with or different concentrations of curcumin(0,6.25,12.5μmol/L),NBD cholesterol and apo-AI.④GW9662(20μmol/L)and curcumin(12.5μmol/L)treated M1 subtype macrophages stimulated by oxLDL or NBD cholesterol and apo-AI.Oil red O and BODIPY staining were used to examine the cholesterol influx/efflux.Real time PCR,ELISA and Western blot were used to examine the expression of ABCA1,CD36,PPARγ,and the expression of inflammation cytokines such as IL-1β,IL-6.ELISA was used to examine the expression of inflammation cytokines TNF-αand MMP-9.Results Curcumin alleviated the inflammation response in M1 macrophage and M1 macrophage induced by oxLDL(P<0.05).Curcumin increased the cholesterol influx/efflux in M1 macrophage induced by oxLDL or NBD cholesterol(30 mg/L)and apo-AI(P<0.05).Curcumin up-regulated the expression of PPARγand CD36 in M1 macrophage induced by oxLDL(P<0.05).Curcumin up-regulated the expression of ABCA1,and down-regulated the expression of TNF-αand MMP-9 in M1 macrophage induced by NBD cholesterol(30 mg/L)and apo-AI(P<0.05).Conclusion Curcumin can increase the ability of M1 macrophages to handle harmful lipids,thus promoting lipid processing,disposal and removal,and curcumin can relieve the inflammation response in M1 macrophage induced by LPS and IFN-γ.
作者
陈方圆
田刚
郭宁
周娟
李娟利
袁祖贻
CHEN Fangyuan;TIAN Gang;GUO Ning;ZHOU Juan;LI Juanli;YUAN Zuyi(Department of Cardiovascular Medicine,First Affiliated Hospital of Medical College,Xi’an Jiaotong University,Xi’an 710061,China)
出处
《山西医科大学学报》
CAS
2019年第10期1384-1391,共8页
Journal of Shanxi Medical University
基金
陕西省科学技术厅社会发展基金资助项目(2019SF-114)
作者简介
通讯作者:陈方圆,女,1982-06生,博士,主治医师,E-mail:chenfy-608@163.com。
