Background:NOD-like receptor family CARD domain containing 3(NLRC3)plays an important role in both innate and adaptive immunity.This study was to explore the function and related mechanisms of NLRC3 in a hypoxia/reoxy...Background:NOD-like receptor family CARD domain containing 3(NLRC3)plays an important role in both innate and adaptive immunity.This study was to explore the function and related mechanisms of NLRC3 in a hypoxia/reoxygenation(H/R)-induced inflammatory response in RAW264.7 cells.Methods:Liver ischemia-reperfusion(I/R)model in mice and H/R model in RAW264.7 cells were constructed.Western blotting was used to determine the protein expression level of NLRC3 in liver tissue and NLRC3,TRAF6,p–p65,p65,IκB–α,and the K63-linked ubiquitination level of TRAF6 in cells.The immunofluorescence assay was performed to evaluate the nuclear level of the NF–κB(p65).ELISA was conducted to measure the content of IL–1βin serum and cell supernatant.The interaction between NLRC3 and TRAF6 in cells was analyzed by the Co-IP assay.Results:The NLRC3 protein level in liver tissue was decreased with the prolongation of reperfusion time(P<0.05).The expression of NLRC3 and IκB–αprotein in RAW264.7 was decreased gradually,while the expression of p–p65 and TRAF6 proteins and K63-linked ubiquitination of TRAF6 were increased gradually with the prolongation of reoxgenation time(P<0.05).The Co-IP assay revealed that NLRC3 and TRAF6 can bind to each other directly.However,NLRC3 had no effect on the expression of TRAF6 protein.The ubiquitination test results showed that the K63-linked ubiquitination level of TRAF6 in H/R+Lv–NLRC3 group was significantly lower than that in the H/R+negative control(NC)group(P<0.05).Moreover,the activation of NF–κB in H/R+Lv–NLRC3 group was inhibited compared with that in the H/R+NC group,and the level of the inflammatory factor IL–1βin the cell culture supernatant was also decreased accordingly(P<0.05).Conclusions:NLRC3 might alleviate H/R-induced inflammation in RAW264.7 cells by inhibiting K63-linked ubiquitination of TRAF6.展开更多
NOD样受体家族胱天蛋白酶激活和募集结构域蛋白3(NOD-like receptor family with a caspase activation and recruitment domain containing 3,NLRC3)作为NOD样受体中的负向调控分子,在免疫应答的抗原识别提呈阶段、淋巴细胞增殖活化阶...NOD样受体家族胱天蛋白酶激活和募集结构域蛋白3(NOD-like receptor family with a caspase activation and recruitment domain containing 3,NLRC3)作为NOD样受体中的负向调控分子,在免疫应答的抗原识别提呈阶段、淋巴细胞增殖活化阶段以及免疫效应阶段通过p38信号分子、NF-κB-活化T细胞核因子5复合物(NF-κB-nuclear factor of activated T cells 5 complex,NF-κB-NFAT5)和丝裂原活化蛋白激酶/细胞外调节蛋白激酶(mitogen-activated protein kinase/extracellular regulated protein kinase,MEK/ERK)信号通路等多种机制发挥抑制作用,在诸多免疫相关疾病,包括感染性疾病、自身免疫性疾病和肿瘤等疾病进程中具有重要调控作用。本文综述了NLRC3分子在不同免疫应答阶段和免疫相关疾病中的作用及机制。展开更多
基金This study was supported by grants from the National Natural Science Foundation of China(81873592)the graduate tutor team construction project of Chongqing Municipal Education Commission Foundation(dstd201801).
文摘Background:NOD-like receptor family CARD domain containing 3(NLRC3)plays an important role in both innate and adaptive immunity.This study was to explore the function and related mechanisms of NLRC3 in a hypoxia/reoxygenation(H/R)-induced inflammatory response in RAW264.7 cells.Methods:Liver ischemia-reperfusion(I/R)model in mice and H/R model in RAW264.7 cells were constructed.Western blotting was used to determine the protein expression level of NLRC3 in liver tissue and NLRC3,TRAF6,p–p65,p65,IκB–α,and the K63-linked ubiquitination level of TRAF6 in cells.The immunofluorescence assay was performed to evaluate the nuclear level of the NF–κB(p65).ELISA was conducted to measure the content of IL–1βin serum and cell supernatant.The interaction between NLRC3 and TRAF6 in cells was analyzed by the Co-IP assay.Results:The NLRC3 protein level in liver tissue was decreased with the prolongation of reperfusion time(P<0.05).The expression of NLRC3 and IκB–αprotein in RAW264.7 was decreased gradually,while the expression of p–p65 and TRAF6 proteins and K63-linked ubiquitination of TRAF6 were increased gradually with the prolongation of reoxgenation time(P<0.05).The Co-IP assay revealed that NLRC3 and TRAF6 can bind to each other directly.However,NLRC3 had no effect on the expression of TRAF6 protein.The ubiquitination test results showed that the K63-linked ubiquitination level of TRAF6 in H/R+Lv–NLRC3 group was significantly lower than that in the H/R+negative control(NC)group(P<0.05).Moreover,the activation of NF–κB in H/R+Lv–NLRC3 group was inhibited compared with that in the H/R+NC group,and the level of the inflammatory factor IL–1βin the cell culture supernatant was also decreased accordingly(P<0.05).Conclusions:NLRC3 might alleviate H/R-induced inflammation in RAW264.7 cells by inhibiting K63-linked ubiquitination of TRAF6.
文摘NOD样受体家族胱天蛋白酶激活和募集结构域蛋白3(NOD-like receptor family with a caspase activation and recruitment domain containing 3,NLRC3)作为NOD样受体中的负向调控分子,在免疫应答的抗原识别提呈阶段、淋巴细胞增殖活化阶段以及免疫效应阶段通过p38信号分子、NF-κB-活化T细胞核因子5复合物(NF-κB-nuclear factor of activated T cells 5 complex,NF-κB-NFAT5)和丝裂原活化蛋白激酶/细胞外调节蛋白激酶(mitogen-activated protein kinase/extracellular regulated protein kinase,MEK/ERK)信号通路等多种机制发挥抑制作用,在诸多免疫相关疾病,包括感染性疾病、自身免疫性疾病和肿瘤等疾病进程中具有重要调控作用。本文综述了NLRC3分子在不同免疫应答阶段和免疫相关疾病中的作用及机制。
