首页|脂多糖激活LRG1/ROCK1信号轴调控中性粒细胞炎症反应

脂多糖激活LRG1/ROCK1信号轴调控中性粒细胞炎症反应

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目的 探讨脂多糖(lipopolysaccharide,LPS)调控富含亮氨酸 α-2 糖蛋白 1(leucine-rich α-2 glycoprotein 1,LRG1)/Rho相关蛋白激酶(Rho-associated protein kinase,ROCK1)信号轴在中性粒细胞炎症反应中的作用.方法 1 μmol/L 全反式维甲酸(all-trans retinoic acid,ATRA)和 12.5 mL/L 二甲基亚砜(dimethyl sulfoxide,DMSO)联合诱导人早幼粒白血病HL-60细胞72 h和96 h后,Wright-Giemsa染色观察细胞形态学变化;流式细胞术检测细胞表面分化抗原CD11b的表达变化.LPS诱导dHL-60细胞和健康人外周血中性粒细胞活化,qPCR和ELISA法分别检测LRG1、ROCK1以及炎性因子的转录和分泌水平;采用Western blotting法检测dHL-60细胞活化后LRG1、ROCK1的蛋白表达;使用重组蛋白LRG1和ROCK1抑制剂Y-27632处理dHL-60细胞,qPCR检测ROCK1和炎性因子的转录水平.结果 在LPS刺激下,中性粒细胞活化,LRG1、ROCK1表达水平显著增加,伴随炎性因子转录水平明显升高;使用重组蛋白LRG1体外刺激dHL-60细胞,ROCK1及炎性因子的转录显著上调;使用ROCK1抑制剂Y-27632,炎性因子水平显著降低.结论 LPS可激活LRG1/ROCK1信号轴促进中性粒细胞炎性因子的生成,加剧炎症反应.
Lipopolysaccharide regulates neutrophil inflammation through activating the LRG1/ROCK1 signaling
Objective To investigate the role of lipopolysaccharide(LPS)in regulating the inflammatory response of neutrophil through the leucine-rich α-2 glycoprotein 1(LRG1)/Rho-associated protein kinase(ROCK1)signaling.Methods HL-60 cells were treated with 1 μmol/L all-trans retinoic acid(ATRA)and 12.5 μL/mL dimethyl sulfoxide(DMSO)for 72 h and 96 h,and the morphological changes were observed by Wright-Giemsa staining.The expression of CD11b was detected by flow cytometry.LPS induced the activation of dHL-60 and human peripheral blood neutrophils.The transcription and secretion levels of LRG1,ROCK1 and inflammatory cytokines were detected by qPCR and ELISA,respectively.The expression levels of LRG1 and ROCK1 after the activation of dHL-60 were detected by Western blotting.Furthermore,dHL-60 was treated with the recombinant protein LRG1 and ROCK1 inhibitor Y-27632;the transcription levels of inflammatory cytokines were detected by qPCR.Results Neutrophils were activated by LPS.The expression levels of LRG1 and ROCK1 were significantly increased,and the transcription levels of inflammatory cytokines were significantly increased.The recombinant protein LRG1 activated dHL-60 in vitro,and the transcription levels of ROCK1 and inflammatory cytokines were significantly increased.Using the ROCK1 inhibitor Y-27632,the production levels of inflammatory cytokines were significantly reduced.Conclusion LPS can regulate the production levels of neutrophil inflammatory cytokines through activating the LRG1/ROCK1 signaling,thus exacerbating the inflammatory response.

lipopolysaccharide(LPS)neutrophilLRG1ROCK1inflammatory response

冯巧、韩欣、袁博慧、张雪娇、华慧、程万鹏、秦苏萍、周峰、刘晓梅

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徐州医科大学江苏省免疫与代谢实验室,江苏徐州 221004

脂多糖(LPS) 中性粒细胞 LRG1 ROCK1 炎症反应

国家自然科学基金资助项目江苏省自然科学基金项目徐州市科技创新专项基金项目

81971179BK20231347KC23242

2024

西安交通大学学报(医学版)
西安交通大学

西安交通大学学报(医学版)

CSTPCD北大核心
影响因子:1.144
ISSN:1671-8259
年,卷(期):2024.45(4)