摘要
目的:探讨β-胡萝卜素对脂多糖(LPS)刺激巨噬细胞RAW264.7炎症因子的影响及其机制。方法:采用5μg/ml的LPS刺激巨噬细胞24 h后用不同浓度的β-胡萝卜素(20、40、80、160μmol/L)处理细胞3 h,MTT法测细胞活性,荧光定量PCR测炎症因子IL-1β、IL-6、TNF-αmRNA相对表达量,ELISA测炎症因子IL-1β、IL-6、TNF-α的分泌量,Western blot测NF-κB p65蛋白相对表达量。5μg/ml的LPS和不同浓度的NF-κB抑制剂PDTC(1、5、10μg/ml)共同处理巨噬细胞24 h,Western blot测NF-κB p65蛋白相对表达量,荧光定量PCR、ELISA测炎症因子的表达和分泌。比较LPS+PDTC组和LPS+PDTC+β-胡萝卜素组炎症因子和NF-κB p65蛋白相对表达的变化。结果:与LPS组相比,不同浓度的β-胡萝卜素对LPS刺激的巨噬细胞的细胞活性有提升作用,对炎症因子和NF-κB p65蛋白的表达有抑制作用;抑制NF-κB蛋白的表达可以抑制LPS刺激的巨噬细胞炎症因子的分泌;LPS+PDTC+β-胡萝卜素组对炎症因子的抑制作用比LPS+PDTC组明显,且差异显著(P<0.05),但两组对NF-κB p65蛋白的表达不明显。结论:β-胡萝卜素可以通过抑制NF-κB通路中NF-κB p65蛋白的表达抑制LPS刺激的巨噬细胞炎症因子IL-1β、IL-6、TNF-α的分泌,且此通路不是唯一相关的通路。
Objective ： To investigate the effects and mechanism of F-carotene on inflammatory factors （ IL-1β, IL-6, TNF-α） in LPS-induced RAW264. 7 cells. Methods：Firstly, RAW264. 7 cells of being induced by 4 （5 μg/ml）for 24 h were treated with different concentration of β-carotene （20,40,80,160 μmol/L）for 3 h. The cells viability was measured by MTT, the mRNA relative expression of IL-1β, IL-6, TNF-α was detected by fluorescence quantitative PCR, the secretion capacity of IL-1β, IL-6, TNF-α was detected by ELISA and the protein relative expression of NF-KB p65 protein was measured by Western blot. Secondly, RAW264. 7 cells were induced by LPS（5 μg/ml） and different concentration of PDTC （1,5,10 μg/ml）for 24 h, NF-KB p65 protein was measured by Western blot and inflammatory factors were detected by fluorescence quantitative PCR and ELISA. Finally, compared the changes in the relative expression of inflammatory factors and NF-KB p65 protein between LPS ＋ PDTC group and LPS ＋PDTC ＋ β-carotene group. Results： Compared with the LPS-induced group, β-carotene could increase the cell viability of LPS-induced RAW264. 7 cells and inhibied the relative expression of inflammatory factors and NF-KB p65 protein. Inhibited the relative expression of NF-KB p65 protein could reduce the relative expression of inflammatory factors. Compared with the LPS ＋PDTC group, LPS ＋ PDTC ＋ 13-carotene group could inhibit the relative expression of inflammatory factors significantly（P〈O. 05）. But, there was little difference about the relative expression of NF-KB p65 protein between this two groups. Conclusion ： d3-carotene inhibits the relative expression of inflammatory factors（ IL-1 β, IL-6, TNF- α） in LPS-induced RAW264. 7 cells through inhibition of NF-KB p65 protein in NF-KB pathway,this pathway isn＇t unique.
作者
张晓音
张珊珊
吴旻
吉昱斌
黄羽盛
郑鑫
ZHANG Xiao-Yin ZHANG Shan-Shan WU Min JI Yu-Bin HUANG Yu-Sheng ZHENG Xin(College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2017年第6期838-843,共6页
Chinese Journal of Immunology
基金
吉林省科技厅科技成果转化促进计划(20150307021NY)
