硝苯地平对人牙龈成纤维细胞MMP-1和MMP-2 mRNA表达的影响

Effects of nifedipine on expression of matrix metalloproteinase-1 and matrix metalloproteinase-2 in human gingival fibroblasts

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、48 h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48 h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 m RNA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-I （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 inununocytochemistry. The expression levels of MMP-1 and MMP-2 mRNA in GFs were detected by quantitative Real-Time PCR after cells stimulated by nifedipine （1 μg/ml）, LPS （1 μg/ml） and nifedipine （1 μg/ml） ＋LPS（1 iμg/ml）. Results GFs cultured invitro showed vimentin positive and keratin negative signal. The mRNA expression of MMP-1 and MMP- 2 were significantly down-regulated after 48 h 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.