表皮葡萄球菌诱导乳腺成纤维细胞表型转变的机制研究  

Investigation on mechanism of Stphylococcus epidermidis inducing phenotypic transition of fibroblasts

作　　者：汤琦[1] 周绍强[1] 王建逵[1] 杨庄青[1] 黄云超[2] 

机构地区：[1]昆明医科大学第三附属医院云南省肿瘤医院乳腺二科,云南昆明650118 [2]昆明医科大学第三附属医院云南省肿瘤医院心胸血管外科,云南昆明650118

出　　处：《中华医院感染学杂志》2016年第3期524-527,共4页Chinese Journal of Nosocomiology

基　　金：国家自然科学基金资助项目(81260228);云南省科技厅-昆明医科大学联合专项基金资助项目(2012FB070);云南省肿瘤医院博士科研启动基金资助项目(BSJJ2015050)

摘　　要：目的检测成纤维细胞在表皮葡萄球菌(SEP)刺激下α-平滑肌肌动蛋白(α-SMA)表达水平及细胞上清中转化生长因子β1(TGF-β1)含量的变化,以探讨成纤维细胞在表皮葡萄球菌刺激下发生表型转变为肌成纤维细胞的机制。方法将细胞培养液分为表皮葡萄球菌SEP RP62A组(A组)、SEP ATCC 12228组(B组)及普通培养液组(C组),RT-PCR法检测各组培养液中的成纤维细胞α-SMA蛋白的表达水平,ELISA法检测各组的成纤维细胞上清中TGF-β1的含量。结果 A组细胞的α-SMA蛋白表达水平明显高于B组及普通培养液组,差异有统计学意义(P<0.01);3组细胞的α-SMA蛋白表达水平均于培养第5天升高,与第3天相比差异有统计学意义(P<0.01);A组及B组细胞第7天仍能维持相比第5天较为稳定的α-SMA蛋白表达水平,差异无统计学意义;C组细胞的α-SMA蛋白表达水平则于培养第7天时明显下降(P<0.05);A组细胞上清中TGF-β1的含量均高于B组及普通培养液组,差异有统计学意义(P<0.01),并与细菌裂解液浓度呈正相关;低浓度的B组细胞上清中TGF-β1含量与C组比较,差异无统计学意义,随细菌裂解液浓度增加,与C组相比,差异有统计学意义(P<0.05)。结论表皮葡萄球菌生物膜形成阳性菌株可诱导乳腺成纤维细胞表型转变为肌成纤维细胞,且能力明显强于表皮葡萄球菌生物膜形成阴性菌株,细菌刺激后细胞上清中TGF-β1含量增加,可能是成纤维细胞发生表型转变的机制之一。OBJECTIVE To investigate the expression of alpha-smooth muscle actin（α-SMA）and transform growth factor-beta1（TGF-β1）level in cell supernatants after the activation of fibroblasts（Fb）by Stphylococcus epidermidis（SEP）,so as to observe the mechanism of transition of fibroblasts into myofibroblasts（MFb）activated by SEP.METHODS The cell supernatants were divided into three groups：group SEP RP62A（group A）,group SEP ATCC12228（group B）and control group（group C）.The expressions ofα-SMA of Fb under different culture conditions were determined by RT-PCR and TGF-β1levels in the cell supernatant cultured with different media were determined by ELISA assay.RESULTS RT-PCR showed thatα-SMA expression was significantly higher in group A than in normal control and group B（P〈0.01）.The level ofα-SMA expression in all 3groups of cells increased at five days of culture,with statistical significance compared to the level at three days（P〈0.01）.The groups A and B maintained stable level ofα-SMA expression on the 7th day as the 5th day,without statistical significance.However,α-SMA expression of group C significantly decreased compared to that on the 5th day（P〈0.05）.The TGF-β1level in the supernatant of group A was significantly higher than that of normal control and group B（P〈0.01）,and it was positively correlated with SEP lysate concentration.The TGF-β1level in group B with low lysate concentration had no significant difference compared to that in the normal control group;however,with the increase of lysate concentration,significantly differences were found compared to group C（P〈0.05）.CONCLUSIONBiofilm-positive SEP strains could promote transformation of Fb to MFb,and showed more potent effect than biofilm-negative SEP strains.The increase of TGF-β1levels in the supernatant would be the one of mechanisms of Fb activated to MFb by Stphylococcus epidermidis.

关 键 词：表皮葡萄球菌 成纤维细胞 肌成纤维细胞 Α-平滑肌肌动蛋白 转化生长因子-Β1 

分 类 号：R378.11[医药卫生—病原生物学]