摘要
目的探究肿瘤坏死因子-α(TNF-α)在小鼠利什曼原虫脾脏感染中的作用。方法在小鼠的右侧后足皮下注射利什曼原虫的前鞭毛体建立感染模型。测量小鼠足垫厚度和体质量监测感染;通过HE染色观察感染后脾脏病理学变化;流式细胞术检测脾脏内淋巴细胞和巨噬细胞的改变;间接免疫荧光法检测脾脏内精氨酸酶1(Arg-1)和诱导型一氧化氮合酶(iNOS)的表达水平;体外调控TNF-α观察对巨噬细胞感染利什曼原虫的影响。结果与B6.WT小鼠相比,B6.TNF^(-/-)小鼠感染后42 d脾脏明显肿大,结构紊乱,各种细胞浸润并分散在整个脾脏中。流式结果显示,感染利什曼原虫后,小鼠脾脏中T细胞和B细胞比例未发生明显改变,而CD11b单核细胞明显增加;免疫荧光结果显示,M2型单核/巨噬细胞的标志物Arg-1在B6.TNF^(-/-)小鼠脾脏中高表达(P<0.05);B6.WT小鼠的脾脏中iNOS表达相对较强(P<0.05);体外研究显示,TNF-α缺失或抑制TNF-α后,腹腔巨噬细胞感染利什曼原虫明显增加(P<0.01),而加入TNF-α后显著抑制了该细胞的感染(P<0.01)。结论B6.TNF^(-/-)小鼠后足垫皮下接种利什曼原虫后发生脾脏感染,可能与TNF-α的缺失导致巨噬细胞发生M2型分化和一氧化氮产生减少有关。
Objective To explore the role of tumour necrosis factor-α(TNF-α)in splenic infection of mice by Leishmania major.Methods To establish an infection model,promastigotes of Leishmania were injected intradermally into the right hind foot of mice.The thickness of the footpad and body weight were measured to monitor the infection.Histological changes in the spleen after infection were observed by HE staining.Changes in lymphocytes and monocytes in the spleen were detected by flow cytometry.The expression level of arginase-1(Arg-1)and inducible nitric oxide synthase(iNOS)in the spleen was determined by indirect immunofluorescence.The effect of TNF-αon macrophage infection with Leishmania was evaluated in vitro.Results Compared to B6.WT mice,the spleens of B6.TNF^(-/-)mice showed significant enlargement 42 days post-infection,with structural disruption.Various cells infiltrated and were dispersed throughout the entire spleen.Flow cytometry results indicated that after infection with Leishmania,there was no significant change in the proportions of T cells and B cells in the spleens of the mice,while CD11b monocytic cells significantly increased.Immunofluorescence results revealed that the M2 macrophage/monocyte marker Arg-1 was highly expressed in the spleens of B6.TNF^(-/-)mice(P<0.05).The expression of iNOS in the spleens of B6.WT mice was relatively strong(P<0.05).In vitro studies found that the absence or inhibition of TNF-αsignificantly increased the infection of Leishmania by peritoneal macrophages(P<0.01),while the addition of TNF-αmarkedly inhibited this infection(P<0.01).Conclusion The splenic infection in B6.TNF^(-/-)mice following subcutaneous inoculation of Leishmania in the hind footpad may be associated with the absence of TNF-α,which leads to M2-type differentiation of macrophages and reduced nitric oxide production.
作者
甄二娜
吴余玲
胡姗姗
Zhen Erna;Wu Yuling;Hu Shanshan(Institute of Clinical Pharmcology,School of Pharmaceutical Science,Anhui Medical University,Key Laboratory of Anti-Inflammatory and Immune Medicine,Hefei 230032;Dept of Clinical Laboratory,The First Affiliated Hospital of University of Science and Technology of China,Hefei 230036)
出处
《安徽医科大学学报》
北大核心
2025年第12期2325-2332,共8页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金项目(编号:81703524)。
