MyD88信号通路抑制对M2型巨噬细胞极化及泡型棘球蚴病肝纤维化的影响  

Effect of myeloid differentiation factor-88(MyD88)signaling pathway inhibition on polarization of M2-type macrophages and hepatic fibrosis in alveolar echinococcosis(AE)

作　　者：孟·孟根 马玉玉 朱江 朱时雨 刘玉梅 马秀敏 王亮[1] MENG Menggen;MA Yuyu;ZHU Jiang;ZHU Shiyu;LIU Yumei;MA Xiumin;WANG Liang(The Fifth Clinical Medical College of Xinjiang Medical University;Clinical Laboratory Center,the Affiliated Tumor Hospital of Xinjiang Medical University,Urumqi 830011,China;Department of Abdominal Surgery,the Third People′s Hospital of Xinjiang Uygur Autonomous Region;The Affiliated Traditional Chinese Medicine Hospital of Xinjiang Medical University,Urumqi 830000,China)

机构地区：[1]新疆医科大学第五临床医学院 [2]新疆医科大学附属肿瘤医院临床检验中心,乌鲁木齐830011 [3]新疆维吾尔自治区第三人民医院腹部外科 [4]新疆医科大学附属中医医院,乌鲁木齐830000

出　　处：《新疆医科大学学报》2025年第4期438-446,共9页Journal of Xinjiang Medical University

基　　金：省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2023-MK5);国家自然科学基金项目(82360127)。

摘　　要：目的探讨阻断髓样分化因子-88(Myeloid differentiation factor-88,MyD88)信号通路后泡型棘球蚴病(Alveolar echinococcosis,AE)肝脏组织中巨噬细胞浸润程度、炎症及纤维化的相关性。方法保种沙鼠肝脏组织中获得泡型棘球蚴原头节(Protoscoleces,PSC),肝脏穿刺PSC分别建立泡型棘球蚴感染组(Em组)和MyD88信号通路阻断的泡型棘球蚴小鼠组(Em+TJ-M2010-5组)。通过免疫组织化学(Immunohistochemistry,IHC)和qRT-PCR分析MyD88、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和巨噬细胞极化标志物的表达情况。体外培养小鼠巨噬细胞系RAW264.7,棘球蚴蛋白(Echinococcus-multilocularis-Protein,Emp)刺激RAW264.7后使用TJ-M2010-5阻断MyD88信号通路,通过蛋白印记实验(Western blot,WB)和qRT-PCR检测各组巨噬细胞极化标志物和MyD88的蛋白及mRNA的表达水平,流式细胞术检测阻断MyD88信号通路后M1型巨噬细胞(F4/80++CD86++)、M2型巨噬细胞(F4/80++CD206++)比例动态变化情况。结果与Em组相比,Em+TJ-M2010-5组MyD88 IHC阳性区域面积减少(P<0.001),α-SMA和CD163阳性区域面积增加(P均<0.001),诱导型一氧化氮合酶(Inducible nitric oxide synthase,INOS)和CD86阳性区域面积减少(P均<0.001)。在体外细胞实验中,与空白对照组相比,泡球蚴蛋白刺激后,MyD88通路激活,MyD88和精氨酸酶-1(Arginine-1)蛋白及mRNA表达水平均升高(P<0.01~0.05),TJ-M2010-5干预后,MyD88及INOS的蛋白及mRNA表达水平均下降(P均<0.05),流式细胞术检测M2型巨噬细胞(F4/80++CD206++)比例由(7.76±0.44)%升高为(17.24±0.47)%(P<0.01),M1巨噬细胞(F4/80++CD86++)比例由(42.16±0.56)%降低为(32.35±1.15)%(P<0.01)。结论在泡型棘球蚴病中抑制MyD88信号通路后,巨噬细胞向M2型巨噬细胞方向极化并加重肝脏纤维化。Objective To explore the correlation between macrophage infiltration,inflammation,and fibrosis in liver tissue of alveolar echinococcosis(AE)after blocking the signal pathway of myeloid differentiation factor-88(MyD88).Methods The protoscolex(PSC)of AE was obtained from the liver tissue of gerbils,and the infection model group of AE(Em group)and the mouse model of AE with blocked MyD88 signal pathway(Em+TJ-M2010-5 group)were established by liver puncture PSC respectively.Immunohistochemistry(IHC)and qRT-PCR were used to analyze the expression of MyD88,α-smooth muscle actin(α-SMA)and macrophage polarization marker.The mouse macrophage cell line RAW264.7 was cultured in vitro.After the mouse macrophage cell line RAW264.7 was stimulated by echinococcus multilocularis protein(Emp),the signal pathway of MyD88 was blocked by TJ-M2010-5.The expression levels of macrophage polarization markers,protein and mRNA of MyD88 was detected by Western Blot and qRT-PCR.The dynamic changes of the ratio of M1-type macrophages(F4/80++CD86++)and M2-type macrophages(F4/80++CD206++)were detected by flow cytometry after blocking the MyD88 signaling pathway.Results Compared with Em group,the positive areas of MyD88 IHC in Em+TJ-M2010-5 group were decreased(P<0.001),the positive areas ofα-SMA and CD163 were increased(P all<0.001),and the positive areas of inducible nitric oxide synthase(INOS)and CD86 were decreased(P all<0.001).In vitro cell experiment,compared with blank control group,the protein and mRNA expression levels of MyD88 and Arginine-1(Arg-1)were increased after the stimulation of echinococcus multilocularis protein(P<0.01-0.05),while the protein and mRNA expression levels of MyD88 and INOS were decreased after the intervention of TJ-M2010-5(P all<0.05).The proportion of M2-type macrophages(F4/80++CD206++)was increased from(7.76±0.44)%to(17.24±0.47)%(P<0.01),and the proportion of M1 macrophages(F4/80++CD86++)was decreased from(42.16±0.56)%to(32.35±1.15)%(P<0.01).Conclusion Inhibition of MyD88 signaling pathway in AE resu

关 键 词：纤维化 MyD88/NF-κB 巨噬细胞极化 

分 类 号：R383[医药卫生—医学寄生虫学]