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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。
Abstract：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 of α-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 resulted in polarization of macrophages towards M2 macrophages and increased liver fibrosis.
[1] WOOLSEY I D,MILLER A L.Echinococcus granulosus sensu lato and Echinococcus multilocularis:A review[J].Res Vet Sci,2021,135(1):517-522.
[2] YANG S,WU J,DING C,et al.Epidemiological features of and changes in incidence of infectious diseases in China in the first decade after the SARS outbreak:an observational trend study[J].Lancet Infect Dis,2017,17(7):716-725.
[3] WEN H,VUITTON L,TUXUN T,et al.Echinococcosis:Advances in the 21st Century[J].Clin Microbiol Rev,2019,32(2):e00075-18.
[4] CONRATHS F J,PROBST C,POSSENTI A,et al.Potential risk factors associated with human alveolar echinococcosis:Systematic review and meta-analysis[J].PLos Negl Trop Dis,2017,11(7):e0005781-e0005801.
[5] ZHANG C,WANG H,LI J,et al.Involvement of TIGIT in natural killer cell exhaustion and immune escape in patients and mouse model with liver echinococcus multilocularis infection[J].Hepatology,2021,74(6):3376-3393.
[6] BAKHTIAR N M,SPOTIN A,MAHAMI O M.Recent advances on innate immune pathways related to host-parasite cross-talk in cystic and alveolar echinococcosis[J].Parasit Vectors,2020,13(1):222-232.
[7] LI Y,WEI M,YUAN Q,et al.MyD88 in hepatic stellate cells promotes the development of alcoholic fatty liver via the AKT pathway[J].J Mol Med (Berl),2022,100(7):1071-1085.
[8] BRENNAN J J,GILMORE T D.Evolutionary origins of toll-like receptor signaling[J].Mol Biol Evol,2018,35(7):1576-1587.
[9] WANG H,ZHANG C S,FANG B B,et al.Dual role of hepatic macrophages in the establishment of the echinococcus multilocularis metacestode in mice[J].Front Immunol,2021,11(2):600625-600635.
[10] GE S,YANG W,CHEN H,et al.MyD88 in macrophages enhances liver fibrosis by activation of NLRP3 inflammasome in HSCs[J].Int J Mol Sci,2021,22(22):12403-12413.
[11] 黄红，张淑坤.多房棘球蚴病的浸润和转移机制研究进展[J].中国人兽共患病学报，2016,32(7):670-673.
[12] 谭雅超，李亮，李智德，等.泡球蚴感染小鼠肝脏α-SMA和Ⅰ型胶原的表达及其意义[J].中国病原生物学杂志，2020,15(11):1324-1327.
[13] 汲蕊，梁瑞文，管志玉，等.TLR4/NF-κB信号通路在微小隐孢子虫感染中的作用机制[J].中国寄生虫学与寄生虫病杂志，2018,36(4):361-365.
[14] WANG S,ZHANG Z,WANG Y,et al.Toxoplasma gondii excretory/secretory antigens suppress pro-inflammatory cytokine secretion by inhibiting TLR-induced NF-κB activation in LPS stimulated murine macrophages[J].Oncotarget,2017,8(51):88351-88359.
[15] 李树志，刘铁军.基于TLR4/MyD88/NF-κB信号通路探讨毒消肝清丸对肝硬化内毒素血症大鼠肝组织炎症反应的影响[J].临床肝胆病杂志，2019,35(9):1958-1964.
[16] 苗娜，王志强，臧美玲，等.TLR4、MyD88在泡状棘球蚴感染小鼠中的表达水平及机制[J].诊断病理学杂志，2022,29(1):37-40.
[17] ZHANG C,LIN R,LI Z,et al.Immune exhaustion of T cells in alveolar echinococcosis patients and its reversal by blocking checkpoint receptor TIGIT in a murine model[J].Hepatology,2020,71(4):1297-1315.
[18] CHAI H,LEI Z,LIU Y,et al.miR-505-5p alleviates acute rejection of liver transplantation by inhibiting MyD88 and inducing M2 polarizationof Kupffer cells[J].Acta Biochim Biophys Sin,2022,54(8):1148-1158.
[19] KADOMOTO S,IZUMI K,MIZOKAMI A.Macrophage polarity and disease control[J].Int J Mol Sci,2021,23(1):134-144.
[20] FAIRFAX K,NASCIMENTO M,HUANG S C.Th2 responses in schistosomiasis[J].Semin Immunopathol,2012,34(6):863-871.
[21] DEWALS B G,MARILLIER R G,HOVING J C.IL-4Ralpha-independent expression of mannose receptor and Ym1 by macrophages depends on their IL-10 responsiveness[J].PLos Negl Trop Dis,2010,4(5):e678-e689.
[22] BINATTI E,GERUSSI A,BARISANT D,et al.The role of macrophages in liver fibrosis:New therapeutic opportunities[J].Int J Mol Sci,2022,23(12):6639-6649.
[23] ZHANG X,YANG X,ZHANG S,et al.Wei-Tong-Xin exerts anti-inflammatory effects through TLR4-mediated macrophages M1/M2 polarization and affects GLP-1 secretion[J].J Pharm Pharmacol,2023,75(4):574-584.
[24] KANG Y F,BAI X,WANG K Y,et al.Zhilong Huoxue Tongyu Capsule regulates the macrophage polarization and inflammatory response via the let-7i/TLR9/MyD88 signaling pathway[J].J Ethnopharmacol,2024,330(2):118107-118208.
[25] MA Y,LI J,LIU Y.Identification and exploration of a new M2 macrophage marker MTLN in alveolar echinococcosis[J].Int Immunopharmacol,2024,131(3):111788-111808.
[26] LI Y Q,CHEN Y F,DANG Y P,et al.Corilagin counteracts IL-13Rα1 signaling pathway in macrophages to mitigate schistosome egg-induced hepatic fibrosis[J].Front Cell Infect Microbiol,2017,7(3):434-443.
基本信息:
中图分类号:R532.32
引用信息:
[1]孟·孟根,马玉玉,朱江,等.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[J].新疆医科大学学报,2025,48(04):438-446.
基金信息:
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)
2025-04-15
2025-04-15