HIF-1α对CIA小鼠胸腺巨噬细胞的影响及RAW264.7巨噬细胞的调控作用  

Effect of HIF-1α on thymic macrophages in CIA mice and its regulation on RAW264.7 macrophages

作　　者：高佩媛 庞春艳[1] 张文兰[1] GAO Peiyuan;PANG Chunyan;ZHANG Wenlan

机构地区：[1]内蒙古科技大学包头医学院第一附属中心实验室(内蒙古自体免疫学重点实验室),内蒙古包头014010

出　　处：《包头医学院学报》2025年第4期23-28,33,共7页Journal of Baotou Medical College

基　　金：内蒙古自然科学基金(2019LH08043)。

摘　　要：目的:探讨低氧诱导因子1-α(hypoxiainduciblefactor-1α,HIF-1α)对巨噬细胞株RAW264.7和胶原诱导关节炎(collagen induced arthritis,CIA)小鼠胸腺组织中巨噬细胞调控作用。方法:(1)RAW264.7巨噬细胞分为空白组、阴性对照组、HIF-1α-siRNA组和过表达HIF-1α组,RT-PCR和免疫蛋白印迹(westernblot,WB)检测HIF-1α在mRNA和蛋白水平的表达情况。RT-PCR检测4个组细胞中CD206、Arg的相对表达量;流式细胞术检测CD16/32^(+)和CD206^(+)巨噬细胞的比例。(2)DBA/1小鼠采用牛Ⅱ型胶原建立CIA小鼠模型,实验分为空白组、模型组、阴性对照组、HIF-1α-siRNA组、过表达HIF-1α组。流式细胞术检测小鼠胸腺中F4/80^(+)CD16/32^(+)M1和F4/80^(+)CD206^(+)M2型巨噬细胞的比例;HE染色观察小鼠踝关节的病理变化。结果:(1)沉默HIF-1α可以使小鼠胸腺组织中F4/80^(+)CD16/32^(+)M1巨噬细胞比例下降,使RAW264.7细胞株和小鼠胸腺组织中F4/80^(+)CD206^(+)M2巨噬细胞比例升高;而过表达HIF-1α作用则相反。(2)CIA小鼠踝关节的滑膜增生,有大量细胞浸润,HIF-1α-siRNA可以减少滑膜组织中淋巴细胞的浸润,减轻滑膜的增生。过表达HIF-1α有相反的作用。结论:HIF-1α可以调节RAW264.7细胞株中巨噬细胞的分化以及调控CIA小鼠胸腺组织中巨噬细胞M1型和M2型的转化,从而影响CIA小鼠的关节炎症进展,说明HIF-1α可以通过调控巨噬细胞的极化而介导CIA小鼠的疾病进程。Objective:To investigate the regulatory effect of hypoxia inducible factor-1α(HIF-1α)on macrophages in the thymus tissue of macrophage cell line RAW264.7 and collagen induced arthritis(CIA)mice.Methods:(1)RAW264.7 macrophages were divided into blank group,negative control group,HIF-1α-siRNA group and overexpression HIF-1α group.RT-PCR and Western blot(WB)were used to detect the expression of HIF-1αat mRNA and protein levels.The relative expression of CD206 and Arg in the four groups of cells was detected by RT-PCR.The proportion of CD16/32^(+)and CD206^(+)macrophages was detected by flow cytometry.(2)CIA mouse model was established by bovine typeⅡcollagen in DBA/1 mice.The experiment was divided into blank group,model group,negative control group,HIF-1α-siRNA group and overexpressed HIF-1α group.The proportion of F4/80^(+)CD16/32^(+)M1 and F4/80^(+)CD206^(+)M2 macrophages in mouse thymus was detected by flow cytometry.HE staining was used to observe the pathological changes of ankle joint in mice.Results:(1)Silencing HIF-1α could decrease the proportion of F4/80^(+)CD16/32^(+)M1 macrophages in mouse thymus tissue,and increase the proportion of F4/80^(+)CD206^(+)M2 macrophages in RAW264.7 cell line and mouse thymus tissue.Overexpression of HIF-1α had the opposite effect.(2)The synovium of the ankle joint of CIA mice was proliferated,with a large number of cell infiltration.While,HIF-1α-siRNA could reduce the infiltration of lymphocytes in the synovial tissue and reduce the proliferation of the synovial membrane.Overexpression of HIF-1α has the opposite effect.Conclusion:HIF-1α can regulate the differentiation of macrophages in RAW264.7 cell line and the transformation of M1 and M2 macrophages in thymus tissue of CIA mice,thus affecting the progress of joint inflammation in CIA mice,indicating that HIF-1α can mediate the disease process of CIA mice by regulating the polarization of macrophages.

关 键 词：低氧诱导因子-1Α 胶原诱导关节炎小鼠 RAW264.7巨噬细胞 

分 类 号：R392.6[医药卫生—免疫学]