硝苯地平对人牙龈成纤维细胞MMP-1和 MMP-2 mRNA表达的影响

目的探讨硝苯地平(Nifedipine)对人牙龈成纤维细胞(gingival fibroblasts, GFs)基质金属蛋白酶-1(matrix metalloproteinase-1, MMP-1)和基质金属蛋白酶-2(MMP-2)mRNA表达的影响。方法 取牙周手术切除的健康牙龈组织,组织块法分离及培养获得GFs,免疫细胞化学法对GFs进行鉴定。对GFs进行药物刺激,分为硝苯地平组(1μg/ml)、LPS组(1μg/ml)、硝苯地平+LPS组(分别为1μg/ml),同时设空白对照组。实时定量PCR检测各组24、48h时MMP-1和MMP-2 mRNA的表达水平。结果 组织块法获得的GFs生长状态良好；免疫细胞化学显示,GFs抗波形丝蛋白染色阳性,抗角蛋白染色阴性。与空白对照组相比,硝苯地平组刺激48h后MMP-1、MMP-2 mRNA表达减少(P＜0.05)；硝苯地平+LPS组MMP-1 mRNA表达明显增加(P＜0.05),MMP-2 mRNA表达无明显变化。结论 硝苯地平可能通过影响GFs MMP-1和MMP-2基因的表达而影响牙龈增生。
Objective To investigate the effects of nifedipine on the expression of matrix metalloproteinase-1(MMP-1) and matrix metalloproteinase-2(MMP-2) in human gingival fibroblasts (GFs) in vitro. Methods The gingival tissue specimens obtained from periodontal surgery were cultured in vitro. GFs were identified by immunocytochemistry. The expression levels of MMP-1 and MMP-2 mRNA in GFs were detected by quantitative Real-Time PCR after cells stimulated by nifedipine(1μg/ml), LPS (1μg/ml) and nifedipine (1μg/ml)+LPS(1μg/ml). ResultsGFs cultured in vitro showed vimentin positive and keratin negative signal. The mRNA expression of MMP-1 and MMP-2 were significantly down-regulated after 48h treatment of nifedipine. The mRNA expression of MMP-1 was significantly up-regulated after GFs were treated with nifedipine+LPS, whereas there were no significant changes in MMP-2 expression. Conclusion These results suggest that nifedipine may be involved in proliferation of gingival fibroblasts through inhibition of MMP-1 and MMP-2 expression