硝苯地平对人牙龈上皮细胞bcl-2基因表达的影响

Nifedipine regulated expression of bcl-2 in human gingival epithelial cells in vitro

目的:体外观察硝苯地平(nifedipine,NIF)对人牙龈上皮细胞(human gingival epithelial cells,HGECs)bcl-2基因转录水平的调节,探讨NIF诱导的药物性牙龈增生(drug-induced gingival overgrowth,DGO)与凋亡抑制基因bcl-2的相关性。方法:采用牙周手术切除的健康牙龈组织,用酶消化法分离培养HGECs;免疫组织化学方法对培养细胞进行细胞鉴定;实时定量PCR技术检测不同浓度NIF(1μg/ml、2μg/ml和3μg/ml)刺激下HGECs中bcl-2 mRNA水平,以0μg/mlNIF为空白对照。采用SPSS 11.0软件包对所得数据进行单因素方差分析。结果:酶消化法获得的HGECs在体外培养中生长状态良好;免疫组织化学显示,HGECs抗角蛋白染色阳性,抗波形蛋白染色阴性;NIF处理24h后的HGECsbcl-2 mRNA水平随NIF浓度的增高而上升,3μg/ml浓度组与空白对照组有显著差异(P<0.05);NIF处理48h后,2μg/ml、3μg/ml浓度组HGECs bcl-2 mRNA水平与空白对照组差异明显(P<0.05)。结论:NIF调节体外培养的HGECs中bcl-2基因转录的水平。

PURPOSE： To investigate the effect of nifedipine（NIF） on transcription of bcl-2 in human gingival epithelial cells （HGECs） in vitro and to study the pathogenesis of epithelial thickening in drug-induced gingival overgrowth （DGO）. METHODS： The gingival tissues obtained from periodontal surgeries were digested with enzyme and HGECs were cultured in vitro; HGECs were identified by immunohistochemistry; bcl-2 mRNA levels were quantitated by Real-time PCR 24 hours and 48 hours after cells stimulated by NIF with different concentration （0μg/ml, 1μg/ml,2μg/ml,3μg/ml）, in which 0μg/ml NIF as blank control. The data was analyzed by one-way ANOVA using SPSS 11.0 software package. RESULTS： HGECs cultured in vitro showed keratin positive signal and vimentin negative signal; the level of bcl-2 mRNA increased with NIF 3μg/ml after 24 hours treatment, which appeared significant increase compared with blank control （P〈0.05）; after 48 hours treatment the level of bcl-2 mRNA in the groups of 2μg/ml and 3μg/ml showed significant increase compared with blank control （P〈0.05）. CONCLUSION： NIF regulates the level of bcl-2 mRNA.