PGRN缺失型腹膜巨噬细胞对细菌脂多糖的体外炎症应答  

Inflammatory responses of PGRN-deficient peritoneal macrophage to bacterial lipopolysaccharide in vitro

作　　者：刘露[1] 张雯[1] 陈翰祥[1] 郑琳[1] 卢翌[2] 王红[1] 唐伟[1] 赵蔚明[1] 

机构地区：[1]山东大学医学院微生物学教研室,济南250012 [2]山东大学医学院生物化学与分子生物学研究所,济南250012

出　　处：《山东大学学报（医学版）》2013年第3期58-62,共5页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(81102229;81271853);山东大学自主创新基金交叉学科基金项目(2011JC008)

摘　　要：目的观察颗粒蛋白前体(PGRN)对细菌脂多糖(LPS)诱导腹膜巨噬细胞(PM)炎症应答的影响。方法诱导提取野生型(WT)小鼠及PGRN基因敲除小鼠(KO)腹膜细胞(PEC),观察PEC数目、形态和类型;流式细胞术检测PEC的巨噬细胞表面标志物CD11b、F4/80。LPS分别处理WT或KO小鼠PM,LPS、重组PGRN或LPS加重组PGRN分别处理WT小鼠PM,培养24 h后收集细胞上清,ELISA法检测肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素12(IL-12),Griess法检测一氧化氮(NO)。结果 PGRN缺失对小鼠PEC的数目、成分及巨噬细胞表面标志物CD11b、F4/80的表达无明显影响;与WT组相比,LPS诱导PGRN KO来源的PM分泌较高水平的TNF-α、IL-1β、IL-12及NO。外源给予重组PGRN后,LPS诱导WT组PM分泌TNF-α、IL-1β、IL-12及NO的水平降低。结论 PGRN缺失使PM对LPS的刺激产生了更强的炎症反应;外源性给予重组PGRN则可抑制LPS诱导PM释放前炎性因子TNF-α、IL-1β、IL-12及炎症介质NO。Objective To investigate the effects of progranulin （PGRN） in the inflammatory responses of peritoneal macrophages （PMs） to bacterial lipopolysaccharide （LPS） in vitro. Methods Peritoneal exudate cells （PECs） were induced and extracted from wild-type （WT） mice and PGRN gene knock-out mice （KO）, then the number, morphology and classes of PECs were subsequently evaluated. Surface markers CD11b and F4/80 of PMs were tested by flow cytometry. PMs derived from WT or KO mice were treated with LPS and WT PMs were treated with PBS, LPS, recombinant PGRN or LPS plus recombinant PGRN respectively. Supernatants of cultivation were collected after 24-hours incubation and concentrations of TNF-α, IL-1β, IL-12 and production of NO were detected by ELISA or Griess assay respectively. Results There were no significant differences in cell number, classes and expression of surface makers CD11b and F4/80 between WT and KO mice-derived PECs. Higher concentration of TNF-α, IL-1β, IL-12 and more NO production were detected in the supernatants of KO PMs stimulated by LPS compared to those of WT PMs. Additionally,recombinant PGRN dramatically inhibited the production of pro-inflammatory cytokines TNF-α, IL-1β, IL-12 and inflammatory intermediate NO of WT PMs stimulated by LPS. Conclusion PGRN KO PMs display a stronger inflammatory response than WT PMs when treated with LPS. In addition, recombinant PGRN powerfully inhibits LPS stimulating production of TNF-α, IL-1β, IL-12 and NO of PMs.

关 键 词：颗粒蛋白前体 腹膜巨噬细胞 基因 PGRN 脂多糖 细胞因子类 小鼠 

分 类 号：R392.12[医药卫生—免疫学]