枸杞多糖促进HL-60细胞的吞噬功能  

Lycium barbarum polysaccharide promotes phagocytic function of HL-60 cells

作　　者：庞洪源 李帆 华梦晴 宋传旺 PANG Hongyuan;LI Fan;HUA Mengqing;SONG Chuanwang(Department of Immunology,School of Laboratory Medicine,Bengbu 233030,China;Anhui Provincial Key Laboratory of Immunology in Chronic Diseases,Bengbu Medical College)

机构地区：[1]蚌埠医学院检验医学院免疫教研室,蚌埠233030 [2]慢性疾病免疫学基础与临床安徽省重点实验室

出　　处：《山西医科大学学报》2024年第1期44-49,共6页Journal of Shanxi Medical University

基　　金：安徽高校自然科学研究项目(KJ2020ZD49);蚌埠医学院“512人才培育计划”项目(by51201103);蚌埠医学院科研创新团体项目(BYKC201902);蚌埠医学院自然科学重点项目(2020byzd024);蚌埠医学院省重点科研平台开放课题(KLICD2022-D2)。

摘　　要：目的探究枸杞多糖(lycium barbarum polysaccharide,LBP)对人中性粒细胞株HL-60细胞吞噬功能的影响及其相关机制。方法采用不同浓度(0,1,10,100,200,500μg/mL)LBP刺激HL-60细胞24 h后,通过CCK-8和流式细胞术检测HL-60细胞的存活率和吞噬功能变化情况,通过免疫印迹法(Western blot)检测HL-60细胞磷脂酰肌醇-3-激酶(PI3K)信号通路Akt磷酸化情况。阻断实验分为对照组、LBP组、LBP+PI3K抑制剂(Wortmannin)组。LBP组以200μg/mL LBP单独刺激HL-60细胞24 h;LBP+Wortmannin组先以1μmol/L Wortmannin预刺激HL-60细胞2 h,再加入200μg/mL LBP继续作用24 h后检测HL-60细胞吞噬功能情况。为了检测HL-60细胞表面受体的表达情况,实验分为对照组和LBP组(200μg/mL),采用流式细胞术检测CD36、CD204、CD209、胶原样结构巨噬细胞受体(macrophage receptor with collagenous structure,MARCO)和Dectin吞噬相关受体表达情况。结果不同剂量LBP处理后,HL-60细胞的存活率未发生明显变化(P>0.05);与0μg/mL LBP比较,1~200μg/mL LBP刺激HL-60细胞吞噬百分率明显增加(P<0.01);与对照组比较,200μg/mL LBP组磷酸化Akt表达增加(P<0.05)。PI3K阻断实验表明,与LBP组比较,LBP+Wortmannin组LBP促进HL-60吞噬E.coli的效应部分抑制(P<0.05)。与对照组比较,LBP组HL-60细胞MARCO受体表达水平明显增加(P<0.05),而CD36、CD204、CD209和Dectin无明显变化(P>0.05)。结论LBP通过PI3K-Akt信号通路促进HL-60细胞吞噬E.coli,这种吞噬功能的提高可能与MARCO受体表达上调有关。Objective To investigate the effect of lycium barbarum polysaccharide(LBP)on phagocytosis of human neutrophil line HL-60 cells and its related mechanism.Methods HL-60 cells were stimulated with different concentrations(0,1,10,100,200,500μg/mL)of LBP for 24 h,and the viability and the phagocytic function of HL-60 cells were detected by CCK-8 and flow cytometry,and the phosphorylation of Akt in HL-60 cells was detected by Western blot.HL-60 cells were divided into control group,LBP group and LBP+PI3K inhibitor(Wortmannin)group for the blocking experiment.HL-60 cells in LBP group were stimulated with LBP(200μg/mL)alone for 24 h.In LBP+Wortmannin group,HL-60 cells were pre-stimulated with 1μmol/L Wortmannin for 2 h,and then treated with 200μg/mL LBP for 24 h.HL-60 cells were divided into control group and 200μg/mL LBP group,and then flow cytometry was used to detect the expressions of CD36,CD204,CD209,macrophage receptor with collagenous structure(MARCO),Dectin,and other receptors.Results Different concentrations of LBP had no significant effect on the survival rate of HL-60 cells(P>0.05).Compared with 0μg/mL LBP,1-200μg/mL LBP significantly increased the phagocytic rate of HL-60 cells(P<0.01).Compared to control group,the expression of phosphorylated Akt increased in 200μg/mL LBP group(P<0.05).The PI3K blocking assay demonstrated that LBP+Wortmannin partially suppressed phagocytosis of E.coli by HL-60 cells induced by LBP(P<0.05).Compared with control group,the expression level of the MARCO receptor in HL-60 cells was significantly increased in LBP group(P<0.05),however,there were no significant changes in CD36,CD204,CD209 and Dectin between two groups(P>0.05).Conclusion LBP promotes the engulfment of E.coli by HL-60 cells through the PI3K-Akt signaling pathway,which may be related to the upregulation of MARCO receptor expression in HL-60 cells.

关 键 词：枸杞多糖 HL-60细胞 吞噬作用 PI3K-AKT信号通路 MARCO受体 

分 类 号：R285[医药卫生—中药学]