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目的探究空肠弯曲菌血红素氧化酶ChuZ对巨噬细胞炎性小体激活的影响。方法分离野生型C57BL/6小鼠骨髓细胞,诱导分化为骨髓来源的巨噬细胞(bone marrow-derived macrophage,BMM),分别用WT C.jejuni、△ChuZ C.jejuni以及对照刺激BMM 6 h,q RT-PCR和ELISA检测各组IL-1β的表达,q RT-PCR和Western blot检测各组NLRP3、ASC、caspase-1的表达。分离野生型C57BL/6和NLRP3-/-小鼠骨髓细胞,诱导分化为WT BMM、NLRP3-/-BMM,分别用WT C.jejuni和△ChuZ C.jejuni刺激BMM 6 h,ELISA检测各组IL-1β的表达。分离野生型C57BL/6小鼠骨髓细胞,诱导分化为BMM,分别用WT C.jejuni、△ChuZ C.jejuni刺激BMM、BMM+MCC950(NLRP3炎性小体抑制剂)6 h,ELISA检测各组IL-1β的表达。结果与对照组相比,WT C.jejuni、△ChuZ C.jejuni均能显著促进BMM IL-1β、NLRP3、procaspase-1、caspase-1的表达(P<0.01);△ChuZ C.jejuni组与WT C.jejuni组相比,IL-1β、NLRP3、procaspase-1、caspase-1的表达显著降低(P<0.01)。与WT BMM组相比,WT C.jejuni、△ChuZ C.jejuni分别刺激NLRP3-/-BMM后,IL-1β表达均显著降低(P<0.01)。同时,与control组相比,WT C.jejuni、△ChuZ C.jejuni刺激+MCC950组后,IL-1β表达均也显著降低(P<0.01)。结论空肠弯曲菌血红素氧化酶ChuZ与巨噬细胞中NLRP3/ASC/caspase-1炎性小体的激活程度相关,并促进巨噬细胞中IL-1β的表达,参与了空肠弯曲菌对宿主的炎性作用。
Objective To investigate the effect of ChuZ of jejunum Campylobacter jejuni on the activation of macrophage inflammasome. Methods Bone marrow cells of wild type C57BL / 6 mice were isolated and differentiated into bone marrow-derived macrophages (BMMs). BMMs were stimulated with WT C.jejuni, ChuZ C. jejuni, and controls for 6 h, The expression of IL-1β in each group was detected by RT-PCR and ELISA. The expression of NLRP3, ASC and caspase-1 in each group was detected by q RT-PCR and Western blot. The bone marrow cells of wild type C57BL / 6 and NLRP3 - / - mice were isolated and induced to differentiate into WT BMM and NLRP3 - / - BMM. BMM was stimulated with WT C.jejuni and ChuZ C. jejuni for 6 h, -1β expression. The bone marrow cells of wild type C57BL / 6 mice were isolated and induced to differentiate into BMM. BMM and BMM + MCC950 (NLRP3 inflammasome inhibitor) were stimulated with WT C.jejuni and ChuZ C.jejuni respectively for 6 h. IL-1β expression. Results Compared with the control group, WT C.jejuni and Chu Chu C.jejuni all significantly promoted the expression of IL-1β, NLRP3, procaspase-1 and caspase-1 in BMM (P <0.01) Compared with WT C.jejuni group, the expression of IL-1β, NLRP3, procaspase-1 and caspase-1 were significantly decreased (P <0.01). Compared with WT BMM group, the expression of IL-1β was significantly decreased after WT C.jejuni and Chu Chu C.jejuni stimulated NLRP3 - / - BMM respectively (P <0.01). At the same time, compared with the control group, the expression of IL-1β was significantly decreased (P <0.01) by WT C.jejuni and △ ChuZ C.jejuni stimulation + MCC950 group. Conclusion The expression of ChuZ in jejunum Campylobacter jejuni correlates with the activation of NLRP3 / ASC / caspase-1 inflammasome in macrophages and promotes the expression of IL-1β in macrophages. Inflammatory effects.