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金黄色葡萄球菌体外感染激活成骨细胞核因子-κB信号通路的研究

Activation of nuclear factor kappaB signaling pathway in human osteoblasts responses to Staphylococcus Aureus in vitro

摘要:

目的 观察金黄色葡萄球菌(简称金葡菌)体外感染激活成骨细胞核因子-κB( NF-κB)信号通路的情况.方法 Western blot检测成骨细胞胞质内核抑制因子-κB (I-κBα)的降解情况,凝胶迁移(EMSA)检测成骨细胞胞核内NF-κB的活性,从而判断成骨细胞NF-κB信号通路的激活情况,ELISA检测应用50 μmol/L SN50预处理1h,再感染1h金葡菌后成骨细胞24 h细胞上清液中IL-6的浓度,观察金葡菌感染所激活成骨细胞的NF-κB信号通路调控免疫反应情况.结果 金葡菌体外感染能够诱导成骨细胞胞质内I-κBα降解(I-κBα15 min/I-κBα0 min=0.409 ±0.245;I-κBα30min/I-κBαmin=0.061 4±0.010)和增加胞核内NF-κB活性,且呈明显时间和剂量依赖性;50 μmol/L SN50预处理后再感染金葡菌的成骨细胞其上清液中IL6的浓度[(2.17 ±0.11) μg/L]相对于单纯金葡菌感染组[(3.58 ±0.31) μg/L]显著减少(F=174.25,P<0.05).结论 金葡菌体外感染能够激活成骨细胞的NF-κB信号通路且其参与调控免疫炎性因子的分泌.

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abstracts:

Objective To investigate whether S.aureus could activate NF-κB signaling pathway in human osteoblasts.Methods Immunoblot and electrophoretic mobility shift assay were used to detect the degradation of I-κBα and activation of NF-κB in human osteoblasts following infection with S.aureus,respectively,and there were investigated the activated state of NF-κB signaling pathway in human osteoblasts.In addition,enzyme-linked immunosorbent assay was used to measure the secretion of IL-6 in culture supernatants,which was represented as one of important cytokines in osteomyelitis,and an inhibitor of NF-κB,SN50,which was added to human osteoblasts culture prior to 1 hour at 50 μmol/L before the infection of S.aureus,was used to determine whether S.aureus -activated NF-κB signaling pathway regulates IL-6 secretion of human osteoblasts.Results S.aureus could induce the degradation of I-κBα ( I-κBα15 min/I-κBα0 min =0.409 ± 0.245 and I-κBα30 min/I-κBα0 min =0.061 ± 0.010) and activation of NF-κB in human osteoblasts in a time and dose-dependent manner following infection.In addition,the secretion of IL-6 in the supernatants of human osteoblasts ( (2.17 ±0.11 ) μg/L) was suppressed by 50 μmol/L SN50 compared to without the addition of SN50 ( ( 3.58 ± 0.31)μg/L) (F=174.25,P<0.05).Conclusions S.aureus could activate NF-κB signaling pathway in human osteoblasts,which could regulate cytokines secretions of human osteoblasts.

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