摘要
目的:探索7-二氟甲氧基-5,4’-二甲氧基金雀异黄素(DFMG)对溶血性磷脂酰胆碱(LPC)处理巨噬细胞(MΦ)增殖活性的影响以及相关信号通路作用机制。方法:首先制备PMA诱导巨噬细胞与LPC诱导巨噬细胞增殖细胞模型,以及不同浓度DFMG孵育后用LPC诱导巨噬细胞增殖,用台盼蓝拒染法和CCK-8法分别检测细胞存活率和增殖活性。ELISA法检测细胞培养液中TNF-α的表达,Western blot检测TRL4、MyD88蛋白表达。结果:1.用150nM PMA佛波酯(PMA)诱导分化为巨噬细胞百分率为(39.67±0.05)%,显著高于未处理组(0.55±0.19)%,用不同浓度LPC(3.0、10.0、30.0、100.0μM)孵育处理巨噬细胞,台盼蓝拒染法和CCK-8法结果测得巨噬细胞增殖率和存活率与空白对照组相比下降均具有统计学意义。故选LPC30.0μM作为LPC处理巨噬细胞模型的最佳造模浓度。2.LPC能诱导巨噬细胞培养液中TNF-α的分泌增加,上调TLR4、MyD88蛋白的表达。3.DFMG以时间和浓度依赖方式增高LPC处理巨噬细胞增殖活性,降低LPC处理巨噬细胞培养液TNF-α浓度,DFMG能拮抗LPC上调巨噬细胞TLR4、MyD88蛋白的表达作用。结论:DFMG能增强LPC对巨噬细胞增殖活性的减弱和降低TNF-α分泌作用与其抑制TLR4-MyD88信号转导相关。
Objective To explore the effects of 7-difluoromethyl-5, 4'-dimethoxygenistein(DFMG) on the proliferation of macrophages treated with LPC and the possible mechanism. Methods The preparation of PMA induced macrophages and macrophage proliferation induced by LPC cell model, and different concentrations of DFMG after incubation with LPC induced macrophage proliferation by trypan blue exclusion and CCK-8 assay were used to detect cell viability and proliferation. ELISA was used to detect the expression of TNF-α in cell culture medium, and Western blotting were used to detect the expression of TRL4 and MyD88 protein. Results 1. The cells differentiation percentage was(39.67±0.05) % after inducing THP-1 cells by 150 nM PMA which was significantly higher than that of control group(0.55±0.19) %. After dealing with different concentrations of LPC(3.0、10.0、30.0、100.0μM), the survival rate and proliferative activity of marcrophages by Typan Blue Exclusion and CCK-8 kits. The difference was statistically significant between cell control group and treatment group.2. After dealing with different concentrations of LPC(3.0、10.0、30.0、100.0μM) can induce the secretion of TNF-alpha in macrophage culture medium and up regulate the expression of TLR4 and MyD88 proteins.3. DFMG can increase the cell proliferative activity in a dose and time-dependent manner. Different concentrations of DFMG(0.3、1.0、3.0μM) can decrease concentrations of TNF-α in the model of macrophages in a dose-dependent manner. DFMG could antagonize the up regulation of TLR4 and MyD88 protein expression in macrophages by LPC. Conclusion DFMG can against the effect of LPC on the inhibiting proliferation of macrophages. DFMG can antagonism the secretion effect of TNF-α in macrophages induced by LPC which may be associated with Toll-like receptor 4-myeloid differentiation factor 88 signal transduction.
出处
《湖南师范大学学报（医学版）》
2018年第1期1-5,共5页
Journal of Hunan Normal University(Medical Sciences)
基金
国家自然科学基金资助项目(No.81370382)
