BRMS1 inhibit IL-1β secretion in synovial cell fibroblasts by suppress the nuclear translocation of NF-κB
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摘要:
目的探讨乳腺癌转移抑制蛋白1（BRMS1）对人类风湿性关节炎（RA）的成纤维样滑膜细胞（FLS）分泌IL-1β的影响和机制。 方法分别取4例RA患者膝关节滑膜组织及1例正常人（normal）膝关节滑膜组织，采用组织贴壁法分离FLS。用慢病毒法过表达BRMS1蛋白。Western blot检测BRMS1蛋白及核蛋白NF-κB水平，Elisa法检测细胞培养基中IL-1β水平，免疫荧光检测NF-κB核转位。 结果RA组FLS的BRMS1蛋白表达较N组明显下调，但NF-κB核转位明显增加、IL-1β分泌明显增加。RA组FLS过表达BRMS1后，FLS核蛋白中NF-κB表达明显减少、IL-1β分泌下降。LPS诱导的NF-κB核转位和IL-1β表达增加被过表达BRMS1明显抑制。 结论BRMS1可以抑制NF-κB核转位，从而减少成纤维样滑膜细胞分泌IL-1β。 -
关键词:
- 类风湿性关节炎 /
- 乳腺癌转移抑制蛋白1 /
- 成纤维样滑膜细胞 /
- NF-kB /
- IL-1β
Abstract:ObjectiveTo investigate the effects and mechanism of breast cancer metastasis suppressor 1(BRMS1) in IL-1β secretion of fibroblast-like synovial cells (FLS) in rheumatoid arthritis (RA). MethodsThe synovial membrane tissues were obtained from 4 RA patients and 1 healthy subject and FLS were isolated by means of tissue culture. FLS were transfected with lenti-virus with BRMS1 protein. BRMS1 protein and nuclear protein NF-κB were detected by Western blot, IL-1β secretion was estimated by Elisa and nuclear translocation of NF-kB was observed by immunofluorescence. ResultsThe BRMS1 protein was obviously decreased in RA group than normal group, but nuclear protein NF-κB and IL-1β secretion were increased significantly. Nuclear protein NF-κB and IL-1β secretion were obviously downregulated in the FLS with BRMS1 protein overexpression of RA group. LPS induced nuclear translocation of NF-κB and IL-1β secretion were obviously inhibited by BRMS1. ConclusionBRMS1 could suppress the nuclear translocation of NF-κB and then reduce the IL-1β secretion of FLS. -
图 1 RA组FLS细胞BRMS1蛋白、核蛋白中NF-kB表达和培养基上清中IL-1β水平的差异
A: Western blot检测总蛋白中BRMS1蛋白水平, 胞浆蛋白中IkKα/β磷酸化水平和IkBα蛋白表达, 核蛋白中NF-κB的表达水平; B: Elisa检测细胞培养基上清中IL-1β水平.与normal组比, *P < 0.05.
图 2 过表达BRMS1对NF-κB核转位和IL-1β表达的影响
A: Western blot检测总蛋白中BRMS1蛋白水平, 胞浆蛋白中IkKα/β磷酸化水平和IkBα蛋白表达, 核蛋白中NF-κB的表达水平; B:免疫荧光检测NF-κB核转位情况。绿色（FITC）为NF-κB, 蓝色（DAPI）为细胞核. C:Elisa检测细胞培养基上清中IL-1β水平.与NC组比, *P < 0.05.
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