沉默Fcgr3降低SiO_(2)诱导的小鼠肺泡巨噬细胞系MH-S炎性因子表达  

Silencing Fcgr3 decreases SiO_(2)-induced inflammatory cytokines expression in mouse alveolar macrophage cell line MH-S

作　　者：李晓娜 齐先梅 张田甜[1] 王婧[1] LI Xiaona;QI Xianmei;ZHANG Tiantian;WANG Jing(Department of Pathophysiology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础学院病理生理学系,北京100005

出　　处：《基础医学与临床》2023年第6期904-908,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(82100080);国家重点研发计划(2021YFC2500700,2021YFC2500704);中央级公益性科研院所基本科研业务费(2021RC310002,2022RC31006)。

摘　　要：目的探讨沉默免疫球蛋白G Fc段受体Ⅲ(FcγRⅢ)对SiO_(2)诱导的小鼠肺泡巨噬细胞系MH-S炎性因子表达的影响。方法构建Fcgr3 shRNA的慢病毒质粒;人胚肾细胞系293T培养包装产生慢病毒;慢病毒感染沉默MH-S中的FcγRⅢ,并筛选稳定感染的细胞。定量多聚酶链反应(qPCR)和蛋白免疫印迹法(Western blot)检测MH-S巨噬细胞系中FcγRⅢ的mRNA和蛋白表达。将MH-S细胞分为MH-S+sh-NC、MH-S+SiO_(2)+sh-NC、MH-S+sh-Fcgr3和MH-S+SiO_(2)+sh-Fcgr3,其中SiO_(2)处理条件为100 mg/L连续刺激12 h。通过qPCR检测炎性因子转录水平,酶联免疫吸附测定(ELISA)检测细胞培养上清中的炎性因子的含量,包括肿瘤坏死因子(TNF-α)、白介素1β(IL-1β)和白介素6(IL-6)。结果包装慢病毒并感染细胞后建立稳定沉默Fcgr3的MH-S细胞系,稳定转染细胞中FcγRⅢ的转录与蛋白表达水平较对照组降低(P<0.05)。与对照组相比,SiO_(2)刺激后Tnf、Il1b和Il6的转录水平明显升高(P<0.05),沉默Fcgr3后炎症因子转录水平显著下降(P<0.05);与对照组相比,SiO_(2)刺激后TNF-α和IL-6的分泌量明显升高(P<0.05),沉默Fcgr3后炎症因子分泌水平显著下降(P<0.05)。结论沉默Fcgr3能够降低SiO_(2)诱导的肺泡巨噬细胞炎性因子的表达。Objective To investigate the effect of receptorⅢfor the Fc region of immunoglobulin G(FcγRⅢ)on SiO_(2)-induced inflammatory factors expression in the mouse alveolar macrophage cell line MH-S.Methods The lentivirus plasmids with Fcgr3 shRNA were constructed;human embryonic kidney cell line 293T was incubated for packaging of the lentivirus for silencing FcγRⅢin MH-S.Quantitative polymerase chain reaction(qPCR)and Western blot were used to detect the transcription level and protein expression of FcγRⅢin MH-S.MH-S cells were divided into MH-S+sh-NC,MH-S+SiO_(2)+sh-NC,MH-S+sh-Fcgr3,MH-S+SiO_(2)+sh-Fcgr3,and the SiO_(2) stimulation condition was incubation in a concentration of 100 mg/L for 12 h.Transcription levels of inflammatory cytokines was detected by qPCR,and the content of inflammatory cytokines in cell culture supernatants were detected by enzyme-linked immunosorbent assay(ELISA),including tumor necrosis factor(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6).Results The stable Fcgr3-silencing cell line was established after lentivirus infection.Transcription level and protein expression of FcγRⅢin stable Fcgr3-silencing cell line were lower than that in control group(P<0.05).The transcription level of Tnf,Il1b and Il6 was significantly increased after SiO_(2) stimulation as compared with the control group(P<0.05),and significantly decreased after Fcgr3 silencing(P<0.05).The secretion of TNF-αand IL-6 was obviously increased after SiO_(2) stimulation(P<0.05),and significantly decreased after Fcgr3 silencing(P<0.05).Conclusions Silencing Fcgr3 may decrease the expression of inflammatory cytokines induced by SiO_(2) in alveolar macrophages.

关 键 词：免疫球蛋白G Fc段受体Ⅲ 巨噬细胞 炎性因子 

分 类 号：R392[医药卫生—免疫学]