苯妥英钠对内毒素作用下人牙周膜成纤维细胞增殖和表达TNF-α的影响

摘要

目的：：观察苯妥英钠( PHT)对内毒素( LPS)作用下人牙周膜成纤维细胞( hPDLFs)增殖和表达肿瘤坏死因子-α( TNF-α)的影响。方法：原代培养 hPDLFs,分别与100 mg/L LPS、不同浓度 PHT (5、20 mg/L)以及100 mg/L LPS+不同浓度PHT共同培养24 h后,采用甲基噻唑基四唑( MTT)法观察不同浓度PHT对LPS作用下hPDLFs增殖活性的影响；采用免疫细胞化学 SP 法检测 PHT 对 LPS 作用下 hPDLFs 内TNF-α的表达情况。结果：100 mg/L 的 LPS 可明显抑制 hPDLFs 的增殖活性、刺激 hPDLFs 表达 TNF-α(P<0.05)；20 mg/L PHT 可明显促进 hPDLFs 的增殖(P <0.05),并可明显抑制 hPDLFs 表达 TNF-α(P<0.05)；将20 mg/L PHT 与100 mg/L LPS 联合作用于 hPDLFs 时,可明显逆转 LPS 对 hPDLFs 增殖(P<0.05),明显拮抗LPS介导hPDLFs表达 TNF-α(P<0.05)。结论：20 mg/L PHT对LPS作用下的hPDLFs具有保护作用。%AIM:To study the effects of phenytoin ( PHT) on the proliferation and tumor necrosis factor-α( TNF-α) expression in lipopolysaccharide ( LPS ) - treated human periodontal ligament fibroblasts ( hPDLFs ) . METHODS:hPDLFs were cultured in vitro and identified by immunocytochemistry ( ICC) SP method. The cells were cultured with 100 mg/L LPS, various concentrations of PHT and 100 mg/L LPS plus various concentrations of PHT re-spectively for 24 h. The cell proliferation was examined by MTT assay. TNF-αexpression in the cytoplasm was detec-ted by SP method. RESULTS:100 mg/L LPS inhibited the proliferation of hPDLFs and increased TNF-αexpression in the cytoplasm of hPDFLs. Phenytoin dose-dependently attenuated the effects of LPS on hPDLFs prolifereation and TNF-α expression. CONCLUSION:PHT may have protective effects on LPS-treated hPDLFs.