结核分枝杆菌RD1蛋白PE35干扰宿主细胞的IL-1β通信  

Mycobacterium tuberculosis region of difference(RD)1 protein PE35 perturbs the IL-1β signaling of host cells

作　　者：冯婧 喻晓雯[1] FENG Jing;YU Xiao-wen(Basic Research Department of Traditional Chinese Medicine&Pharmacy,Chongqing Hospital of Traditional Chinese Medicine,Chongqing 400021,China)

机构地区：[1]重庆市中医院中医药基础研究室,重庆400021

出　　处：《微生物学免疫学进展》2022年第1期15-21,共7页Progress In Microbiology and Immunology

基　　金：重庆市科技局项目(cstc2018jcyjAX0771)。

摘　　要：目的 结核分枝杆菌(Mycobacterium tuberculosis, MTB)PE35(Rv3872)是PE/PPE家族的成员,在多种压力条件下会下调表达。分析其在宿主天然免疫反应中的作用,为结核病的防治提供新思路。方法 将PE35通过脂质体转染进鼠巨噬细胞RAW264.7中,在用脂多糖(lipopolysaccharide, LPS)刺激细胞后,通过RT-PCR和ELISA检测巨噬细胞中细胞因子的表达、Western blot检测巨噬细胞中炎性小体的表达和相关信号通路蛋白的磷酸化状态,探索PE35干扰的信号通路。结果 PE35在RAW264.7细胞中减弱了LPS诱导的白细胞介素-1β(interleukin-1β, IL-1β)的产生和核苷酸结合寡聚化结构域样受体蛋白3(nucleotide binding oligomerization domain like receptors 3, NLRP3)炎性小体的激活(P均<0.05)。PE35降低Akt和IκB-α蛋白的磷酸化(P均<0.05),从而抑制PI3K/Akt和核因子κB(nuclear factor kappa-B, NF-κB)信号通路的活化。结论 PE35通过减弱PI3K/Akt和NF-κB的信号传导来抑制NLRP3激活,从而降低了LPS诱导的巨噬细胞中IL-1β的产生。Objective Mycobacterium tuberculosis PE35(Rv3872), a member of the PE/PPE family which located in RD1, is down-regulated under a variety of stress conditions. We define its role in the host’s innate immune response, providing novel ideas for the prevention and treatment of tuberculosis. Methods PE35 were transfected into RAW264.7 cell by liposome. then RAW264.7 cells were stimulated with LPS and the cytokines expression were detected by RT-PCR and ELISA. They also be examined for the expression of inflammasomes and the phosphorylation status of related signaling pathway proteins in macrophages by western blot, as for determining the signaling pathways interference by PE35. Results PE35 attenuated LPS induced IL-1β(P<0.05) expression and the activation of NLRP3 inflammasome(P<0.05) in macrophages. Moreover, PE35 inhibits the phosphorylation of Akt(P<0.05) and IκB-α(P<0.05), thereby inhibiting the activation of PI3 K/Akt and NF-κB signaling pathways. Conclusion PE35 inhibits NLRP3 activation by attenuating PI3 K/Akt and NF-κB signaling, thereby reducing the production of IL-1β in macrophages induced by LPS.

关 键 词：结核分枝杆菌 PE35 白细胞介素-1Β 核苷酸结合寡聚化结构域样受体蛋白3炎性小体 磷脂酰肌醇3-激酶/Akt丝氨酸/苏氨酸激酶 核因子ΚB 

分 类 号：Q93-33[生物学—微生物学]