摘要
目的:观察细菌内毒素(LPS)对体外培养人皮肤成纤维细胞分泌转化生长因子-β1(TGF-β1)和γ干扰素(IFN-γ)作用的影响,探讨其在增生性瘢痕形成中的可能作用。方法:取增生性瘢痕患者的正常皮肤进行成纤维细胞培养,分别用终浓度为0.005、0.010、0.050、0.100、0.500和1.000μg/ml的大肠杆菌LPS(E.coli055:B5)刺激,传代至表型稳定(第8代)。酶联免疫吸附法测定LPS刺激后8、24和72 h细胞培养上清液中TGF-β1和IFN-γ含量的变化。阴性对照为DMEM培养基;阳性对照为增生性瘢痕成纤维细胞培养上清液。结果:随着LPS浓度的增加(0.005～0.100μg/m1),各时间点细胞培养上清液中TGF-β1的含量增加,而IFN-γ的含量降低,呈明显量-效依赖关系,在0.100μg/ml浓度时作用最为明显,TGF-β1、IFN-γ的含量与阳性对照相近(P>0.05)。但随着浓度的进一步增加(0.500μg/ml),这一作用开始下降,当刺激浓度达到(1.000μg/ml)时,则呈相反作用。结论:一定浓度的LPS刺激可促进成纤维细胞分泌TGF-β1并抑制IFN-γ的分泌,这可能是增生性瘢痕形成的重要机制之一。
Objective：To investigate the effect of lipopolysaccharide （LPS） on the proliferation and excretion of transforming growth factor-β1（TGF-β1） and interferon-γ （IFN-γ） from fibroblast of human skin. Methods：Purified dermal fibroblasts were derived from human normal skin and exposed to different concentrations of LPS （0. 005, 0. 010, 0. 050, 0. 100, 0. 500 and 1. 000 μg/ml）. Cell proliferation was determined by MTT assay. The supernatant was collected at 8, 24 and γ2 hours after LPS stimulated, and TGF-β1, IFN-y were assayed by enzyme-linked immunoadsorbent assay （ELISA）. DMEM cultured fibroblasts served as the negative control while the fibroblasts from hypertrophic scar as the positive control. Results： LPS of lower concentrations （0. 005- 0. 100 μg/ml） enhanced the proliferation and the excretion of TGF-β1 and inhibited the excretion of IFN-γ, and the effect showed a concentration dependent manner. The effect reached the peak when fibroblasts were challenged with LPS in the concentration of 0. 100 μg/ml. When the concentration of LPS was 0. 100 μg/ml, the proliferation and the excretion of TGF-β1, IFN-γ were similar to that of positive control group（P〉0.05）. However, LPS of high concentration （1. 000 μg/ml） showed opposite effect. Conclusion：LPS within certain concentration enhances cell proliferation and excretion of TGF-β1, and inhibits the excretion of IFN-γ. The results suggest that LPS might play an important role in the formation of hypertrophic scar.
出处
《感染．炎症．修复》
2010年第1期10-12,共3页
Infection Inflammation Repair
基金
国家自然科学基金资助项目(30371467)
关键词
脂多糖
成纤维细胞
细胞因子
细胞增殖
Lipopolysaccharide Fibroblast Cytokine Cell proliferation
