骨髓巨噬细胞M1/M2亚群失衡对免疫介导的新型再生障碍性贫血模型小鼠发病的影响  

Role of imbalance of M1/M2 subsets of bone marrow macrophages in the pathogenesis of immuiie-mediated aplastic anemia in mice

作　　者：穆慧 贾惠[1] 林赠华 郑红慧[1] 王莉 刘红 Mu Hui;Jia Hui;Lin Zenghua;Zheng Honghui;Wang Li;Liu Hong(Medical School,Nantong University,Nantong 226001,China;Affiliated Hospital of Nantong University,Nantong 226001,China)

机构地区：[1]南通大学医学院,南通226001 [2]南通大学附属医院,南通226001

出　　处：《中华血液学杂志》2021年第11期945-951,共7页Chinese Journal of Hematology

基　　金：国家自然科学基金(81070400);江苏省"强卫工程"医学重点学科项目(zdxkb2016009);中国博士后科学基金(2020M670038ZX)。

摘　　要：目的探讨巨噬细胞对新型免疫介导的再生障碍性贫血(AA)模型小鼠发病的影响。方法以正常雌性B6D2F1(F1)小鼠作为空白对照(对照组,3只),应用双磷酸盐(Clodronate)脂质体尾静脉注射的方法去除小鼠的巨噬细胞(CLO组,4只),以PBS脂质体处理小鼠为阴性对照(PBS组,4只)。并在注射脂质体后以全身辐照(TBI)联合异基因淋巴细胞输注的方法制备F1 AA小鼠模型,分别记为CLO+AA组和PBS+AA组。骨髓活检病理学检查、外周血细胞计数观察各组小鼠AA的程度;流式细胞术检测小鼠骨髓CD4^(+)/CD8^(+)T淋巴细胞亚群和巨噬细胞亚群;ELISA法检测各组小鼠外周血血浆中IFN-γ、TNF-α、G-CSF、GM-CSF、TPO、EPO的水平变化。结果与PBS+AA组相比,去除巨噬细胞的CLO+AA组小鼠骨髓脂肪化明显改善。PBS+AA组和CLO+AA组的HGB分别为(91.50±31.63)、(110.65±24.15)g/L,PLT分别为(90.85±121.90)×10^(6)/L、(461.13±483.45)×10^(6)/L,差异均有统计学意义(P值均<0.05)。PBS+AA组和CLO+AA组CD4^(+)T淋巴细胞占骨髓细胞的比例分别为(18.50±10.17)%、(7.58±8.00)%;CD8^(+)T淋巴细胞占骨髓细胞的比例分别为(36.23±6.40)%、(6.67±5.78)%。去除巨噬细胞后,CD4^(+)和CD8^(+)T淋巴细胞比值均下降,但CD8+T细胞减少程度更大,差异均有统计学意义(P值均<0.05)。和PBS+AA组相比,CLO+AA组小鼠的脾脏和骨髓巨噬细胞比例均明显减少,M1型巨噬细胞减少程度明显高于M2型,差异均有统计学意义(P值均<0.05)。PBS+AA组和CLO+AA组外周血中IFN-γ水平分别为(602.37±104.62)、(303.01±87.22)ng/L;TNF-α水平分别为(34.46±1.42)、(23.25±4.21)ng/L;GM-CSF水平分别为(9.32±2.00)、(64.85±12.25)ng/L;G-CSF水平分别为(5891.78±2632.39)、(17784.16±488.36)ng/L;EPO水平分别为(9667.31±4501.95)、(2078.02±897.56)ng/L;TPO的浓度分别为(6.36±2.09)、(11.67±2.86)ng/L,差异均有统计学意义(P值均<0.05)。结论该实验证实在新型F1 AA小鼠模型中巨噬细胞参与了AA�Objective To investigate the role of macrophages(M0)in the pathogenesis of modified immune-mediated aplastic anemia(AA)mice model.Methods Before the establishment of the Fl AA mice model by total-body irradiation combined with allogeneic lymphocyte infusion,the mice of the CLO+AA group were treated with clodronate(CLO)liposomes to remove macrophages,and those of the PBS+AA group were treated with phosphate-buffered salinc(PBS)liposomes and used as control.The severity of AA was observed by bone marrow(BM)pathological examination and peripheral blood cell count.Flow cytometry(FCM)was used to detect the CD4^(+)/CD8^(+)T lymphocyte subsets in the BM and Mo subsets in the BM and spleen of each group.The levels of IFN-γ,TNF-α,G-CSF,GM-CSF,EPO,and TPOin the peripheral blood were detected using enzyme-linked immunosorbent assay.Finally,the relationshipsbetween inflammatory factors and Mo subsets were analyzed.Results The BM fatty conversion of mice inthe CLO+AA group was significantly alleviated compared with the PBS+AA group.Hemoglobin countswere(91.50±31.63)and(110.65±24.15)g/L,respectively,and the platclet counts were(90.85±121.90)×10^(6)/L and(461.13±483.45)×10^(6)/L,respectively.The differences were all statistically significant(all P<0.05).After removing macrophages,the proportions of CD4 and CD8’T lymphocytes in BM of mice inthe CLO+AA group decreased,but the reduction of CD8 T cells was more significant.The proportions of CD4^(+) T cells and CD8^(+)T cells in BM of the PBS+AA group were(18.5±10.17)%and(36.23±6.40)%,respectively,and in the CLO+AA group were(7.58±8.00)%and(6.67±5.78)%,respectively.Similarly,the percentage of macrophages in the spleen and BM in the CLO+AA group was significantly reducedcompared with the PBS+AA group,most of which were Ml macrophages(P<0.05).The levels of IFN-γ in peripheral blood of the PBS+AA and CLO+AA groups were(602.37±104.62)ng/L and(303.01±87.22)ng/L,respectively,the levels of TNF-α were(34.46±1.42)ng/L and(23.25±4.21)ng/L,respectively,the levels of GM-CSF

关 键 词：贫血 再生障碍性 模型 动物 巨噬细胞 炎症介导素类 

分 类 号：R556.5[医药卫生—血液循环系统疾病]