Nrf2基因缺陷增强烟雾浓缩物对小鼠的免疫刺激反应—Nrf2基因缺陷影响着小鼠的免疫应激反应

Abstract

目的：分析烟雾浓缩物刺激下的Nrf2基因缺陷的小鼠脾脏淋巴细胞亚群百分比和腹腔巨噬细胞炎症因子变化，初步探讨Nrf2基因缺陷对烟雾浓缩物刺激下小鼠免疫反应的影响。方法：36只雌性C57BL/6 WT和Nrf2-/-小鼠随机分别分为两组；其中两组WT和Nrf2-/-小鼠连续饮用含DMSO溶解烟雾浓缩物的纯净水4个月；另两组小鼠饮用含等比DMSO的纯净水4个月。取小鼠脾脏，拍照，坏死评分；制备单细胞悬液，进行计数；采用流式细胞术分析脾脏淋巴细胞各亚群的变化。同时提取小鼠腹腔巨噬细胞，检测分析相关细胞因子的表达。结果：Nrf2基因缺陷促使了烟雾浓缩物刺激下小鼠脾脏肿大，并伴有坏死，降低了其脾脏总细胞数；增高了烟雾浓缩物刺激下小鼠调节免疫相关的脾脏T淋巴细胞和CD8淋巴细胞的百分比，同时能够降低B淋巴细胞；降低了CD4和CD8中央型记忆T细胞(TCM)或效应性记忆T细胞(TEM)的百分比，增加了效应T细胞(TE)；增强了多数炎症性细胞因子、集落刺激因子和趋化因子在小鼠腹腔巨噬细胞中的表达。结论：Nrf2缺陷增强了烟雾浓缩物激活的小鼠天然免疫和适应性免疫的应答。 Objective: To investigate the variation of spleen lymphocyte subsets percentages and abdominal macrophage inflammatory factor of the cigarette smoke extract-induced Nrf2-/- mice, and to dis-cuss the role of Nrf2-/- in cigarette smoke extract-induced immuno-stimulus response of mice. Methods: 36 female C57BL/6 WT and Nrf2-/- mice were randomly divided into 2 groups, respectively. Two groups of WT and Nrf2-/- mice drank pure water containing DMSO dissolved cigarette smoke extract for 4 months continuously; the other two groups’ mice drank pure water containing the same amount of DMSO for four months. Spleens were harvested for taking pictures and necrosis scores; the single cell suspension of spleen tissue was prepared for splenocyte counting and the variation of spleen lymphocyte subsets was analyzed with flow cytometry. Meanwhile, the abdominal macrophages were harvested for the detecting of the related inflammatory factor. Results: Nrf2-/- prompted the spleen swelling with necrosis and lowered the number of splenocytes of cigarette smoke extract-induced mice; Nrf2-/- increased the percents of immune regulation related spleen total T cells and CD8 T cells and reduced the B cells; Nrf2-/- reduced the CD4 and CD8 TCM or TEM cells and increased the TE cells; Nrf2-/- enhanced the majority of inflammatory cytokines, colony stimulating factors and chemokines expression in abdominal macrophage of cigarette smoke extract-induced mice. Conclusion: Nrf2-/- enhances the cigarette smoke extract-induced natural immune and regulating immune response in mice.