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目的探究减数分裂重组蛋白11同系物A(MRE11)在不同刺激物诱导下炎症小体激活中的作用和地位。方法利用不同刺激物,如Poly(I∶C)、Poly(d A∶d T)、E.coli g DNA、293T g DNA和CPPD以及生殖疱疹病毒(HSV)等处理单核巨噬细胞THP-1,通过IL-1β酶联免疫吸附试剂盒(ELISA)筛选出诱导激活炎症小体的有效刺激物;合成si MRE11干扰小RNA,转染THP-1细胞,免疫印迹检测MRE11干扰下调效率;利用筛选到的阳性刺激物处理MRE11干扰下调的细胞,采用ELISA和免疫印迹方法检测MRE11对细胞分泌炎性因子IL-1β水平和前胱天蛋白酶(pro-caspase)-1切割的影响。结果在MRE11干扰下调的THP-1细胞中,Poly(I∶C)、Poly(d A∶d T)、E.coli g DNA和293T g DNA刺激激活细胞分泌的IL-1β水平以及前胱天蛋白酶-1切割的水平均有不同程度降低。结论MRE11参与由DNA以及RNA刺激激活的炎症小体信号通路。
Objective To investigate the role and location of melanin-11 (MRE11) in the activation of inflammasome under different stimuli. Methods Monocytes were treated with different stimuli such as Poly (I: C), Poly (d A: d T), E. coli DNA, 293T g DNA and CPPD and herpes simplex virus (HSV) 1, IL-1β enzyme-linked immunosorbent assay (ELISA) was used to screen the effective stimuli that induce activation of inflammasome; Synthesized si MRE11 interference small RNA, transfected THP-1 cells, Western Blot to detect MRE11 interference down regulation efficiency; The positive stimulators screened MRE11 downregulation of cells, ELISA and Western blotting method was used to detect MRE11 secretion of inflammatory cytokines IL-1β levels and pro-caspase-1 cleavage. Results Poly (I: C), Poly (dA: dT), E. coli DNA and 293T g DNA stimulated IL-1β levels secreted by activated cells in pre-melangenesis in MRE11-knockdown THP-1 cells, Protease-1 cleavage levels are reduced to varying degrees. Conclusion MRE11 is involved in the inflammasome signaling pathway activated by DNA and RNA.