LPS激活巨噬细胞NLRP3炎性小体的作用研究  被引量:9

Study of LPS-induced activation of NLRP3 inflammasome in macrophages

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作  者:牟娜娜[1] 娄晓盈[1] 王仁庆[1] 谭红梅[1] 

机构地区:[1]中山大学中山医学院病理生理教研室,广东广州510080

出  处:《热带医学杂志》2015年第4期441-443,466,F0003,共5页Journal of Tropical Medicine

基  金:国家自然科学基金(81370371)

摘  要:目的探讨脂多糖(LPS)激活巨噬细胞NLRP3炎性小体的作用。方法用佛波酯诱导人单核细胞(THP-1)分化为巨噬细胞。实验分为对照组、LPS组、LPS+NLRP3 si RNA组、LPS+Scrambled si RNA组。采用免疫荧光染色检测核苷酸结合寡聚化结构域样受体家族pyrin结构域蛋白3(NLRP3)和凋亡相关斑点样蛋白ASC的共定位、Western blotting检测NLRP3蛋白的表达、ELISA检测细胞培养上清白介素1β(IL-1β)的浓度。结果与对照组相比,NLRP3和ASC蛋白的共定位明显增多,LPS组NLRP3蛋白的表达增强。LPS还能使细胞培养上清IL-1β的浓度显著升高,NLRP3 si RNA干扰后细胞培养上清IL-1β的浓度显著降低。结论LPS可以促进炎性小体成分NLRP3、ASC的表达,激活NLRP3炎性小体,刺激巨噬细胞分泌炎症因子IL-1β。Objective This study investigated the lipopolysaccharide(LPS)-induced activation of nucleotide-binding oligomerization domain(NOD)-like receptors 3(NLRP3) in macrophages.Methods Phorbol 12-myristate 13-acetate(PMA)was applied to induce THP-1 cells to differentiate into macrophages.Macrophages were divided into control group(CT),LPS group,LPS+NLRP3 si RNA group and LPS+scrambled si RNA group.Colocalization of NLRP3 with apoptosis-associated specklike protein containing CARD(ASC) was detected by immunofluorescence staining.Expression of NLRP3 protein was detected by Western blotting.Concentration of interleukin-1β in culture supernatant was measured by ELISA.Results Colocalization of NLRP3 with ASC was increased in LPS group compared with CT group.Compared with the control group,the expression of NLRP3 protein increased in LPS group.LPS significantly increased the concentration of interleukin-1β in culture supernatant,which was inhibited by NLRP3 si RNA interference.Conclusion LPS may induce NLRP3 inflammasome formation and activation in macrophages and thereby lead to the secretion of interleukin-1β.

关 键 词:脂多糖 巨噬细胞 炎性小体 NLRP3 IL-1Β 

分 类 号:Q538[生物学—生物化学]

 

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