去分化脂肪细胞对巨噬细胞极化的影响  

Effects of dedifferentiated fat cells on polarization of macrophages

作　　者：刘士强 张钰 熊绍恒 董琛 黄兆松 刘恒鑫 宋雅娟 余州 马显杰 Liu Shiqiang;Zhang Yu;Xiong Shaoheng;Dong Chen;Huang Zhaosong;Liu Hengxin;Song Yajuan;Yu Zhou;Ma Xianjie(Department of Plastic Surgery,the First Affiliated Hospital of Air Force Medical University,Xi’an 710032,China)

机构地区：[1]中国人民解放军空军军医大学第一附属医院整形外科,西安710032

出　　处：《中华实验外科杂志》2022年第2期246-249,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81671925、81971851);陕西省重点研发项目(2018ZDXM-SF-081);西京医院学科助推计划(XJZT18ML44、XJZT19D03)。

摘　　要：目的观察去分化脂肪细胞(DFAT)对巨噬细胞极化的影响。方法分别向培养液中加入脂多糖(LPS)或白细胞介素4(IL-4), 诱导RAW264.7向M1或M2型巨噬细胞极化。分离培养C57BL/6小鼠DFAT, 并对其进行多向分化诱导, 通过油红O染色、茜素红染色进行成脂、成骨干性鉴定, 利用Transwell小室将DFAT与巨噬细胞共培养24 h, 在显微镜下观察巨噬细胞形态变化, 通过实时定量反转录聚合酶链反应(RT-qPCR)检测M1型巨噬细胞相关基因诱导型一氧化氮合酶(iNOS)、人巨噬细胞趋化蛋白-1 (MCP-1)、肿瘤坏死因子α (TNF-α)和M2型巨噬细胞相关基因精氨酸酶-1 (Arg-1)、壳酶蛋白(Ym-1)、白细胞介素-10(IL-10)表达量的变化。组间比较采用t检验。结果加入LPS或IL-4刺激, 可诱导RAW264.7向M1或M2型巨噬细胞极化。将DFAT细胞与巨噬细胞共培养发现, DFAT细胞可使巨噬细胞的形态发生明显变化, M1组和对照组M0组比较, M1型巨噬细胞相关基因TNF-α的表达显著增高(5.02±0.71比1.00±0.02, t=5.648, P<0.01)、iNOS的表达显著增高(7.44±1.56比1.00±0.06, t=4.143, P<0.01)、MCP1的表达显著增高(19.64±2.48比1.00±0.04, t=7.510, P<0.01);M2组M0组比较, M2型巨噬细胞相关基因Arg1的表达增高(165.40±45.72比1.00±0.07, t=3.597, P<0.05)、IL-10的表达增高(6.86±1.91比1.00±0.07, t=3.055, P<0.05)、YM-1的表达增高(5.17±2.27比1.00±0.03, t=1.834, P<0.05);DM1组较于M1组, TNF-α的表达显著降低(0.93±0.40比5.02±0.71, t=4.979, P<0.05)、iNOS的表达显著降低(1.61±0.83比7.44±1.56, t=3.302, P<0.05), MCP1的表达显著降低(2.22±1.27比19.64±2.48, t=6.244, P<0.05);而DM2组与M2组比较, Arg-1的表达显著增高(493.10±55.37比165.40±45.72, t=4.563, P<0.05)、IL-10的表达显著增高(87.84±26.90比6.86±1.92, t=3.002, P<0.05)、YM-1的表达显著增高(49.11±11.07比5.18±2.28, t=3.887, P<0.05)。结论 DFAT细胞可抑制巨噬细胞向M1型巨噬细胞极化, 促进其向M2型巨噬细胞Objective To explore the effect of dedifferentiated fat cells(DFAT)on the polarization of macrophages,and to provide the theoretical support for the application in the treatment of wound.Methods RAW264.7 cells were stimulated with lipopolysaccharides(LPS)or interleukin 4(IL-4)to induce them to polarize to M1 or M2 macrophages.DFAT cells of C57BL/6 mice were cultured,and adipogenic and osteogenic differentiation ability was identified by red O staining and alizarin red staining.After the co-culture of 24-h by Transwell,the morphological changes of macrophages were observed.Real-time quantitative polymerase chain reaction(RT-qPCR)was conducted to detect the mRNA expression of M1 macrophage-related genes,including inducible nitric oxide synthase(iNOS),macrophage chemoattractant protein-1(MCP-1),and tumor necrosis factorα(TNF-α)and M2 macrophage-related genes,including Arginase-1(Arg-1),chitinase-like protein-3(YM-1),and interleukin 10(IL-10).In this experiment,random grouping was used,and comparison between the groups was done by t tests.Results LPS or IL-4 stimulation could induce RAW264.7 polarization to M1 or M2 type macrophages.Co-culture with macrophages and DFAT cells led to significantly morphological changes of macrophages.As compared with control group,the expression of M1 macrophage-related gene TNF-α,iNOS and MCP1 in M0 group was significantly increased(TNF-α:5.02±0.71 vs.1.00±0.02,t=5.648,P<0.01;iNOS:7.44±1.56 vs.1.00±0.06,t=4.143,P<0.01;MCP1:19.64±2.48 vs.1.00±0.04,t=7.510,P<0.01).As compared with M0 group,the expression of M2 macrophage-related genes Arg1,IL-10 and YM-1 was significantly increased in M2 group(Arg1:165.40±45.72 vs.1.00±0.07,t=3.597,P<0.05;IL-10:6.86±1.91 vs.1.00±0.07,t=3.055,P<0.05;YM-1:5.17±2.27 vs.1.00±0.03,t=1.834,P<0.05).Compared with the M1 group,the expression of TNF-α,iNOS and MCP1 in the DM1 group was significantly decreased(TNF-α:0.93±0.40 vs.5.02±0.71,t=4.979,P<0.05;iNOS:1.61±0.83 vs.7.44±1.56,t=3.302,P<0.05;MCP1:2.22±1.27 vs.19.64±2.48,t=6.244,P<0.05).

关 键 词：去分化脂肪细胞 巨噬细胞 极化 免疫调控 

分 类 号：R392[医药卫生—免疫学]