辛伐他汀对人牙周膜成纤维细胞增殖和成骨分化的影响

Effects of simvastatin on the viability and differentiation of human periodontal ligament fibroblasts in vivo.

目的:评价辛伐他汀对人牙周膜成纤维细胞增殖和成骨分化的影响。方法:采用组织块法进行人牙周膜成纤维细胞的分离培养。在培养液中加入不同浓度的辛伐他汀,分为对照组(0 mol/L)和5个实验组(10-8、10-7、10-6、10-5和10-4mol/L),检测各组细胞增殖能力和碱性磷酸酶活性。通过矿化结节茜素红染色和RT-PCR,评价辛伐他汀对细胞成骨能力和成骨相关基因表达的影响。结果:辛伐他汀浓度为10-7mol/L时,细胞增殖能力显著提高(P﹤0.05)。辛伐他汀浓度为10-8、10-7、10-6和10-5mol/L时,细胞碱性磷酸酶活性均有显著增强(P﹤0.05),其中,10-7mol/L组增强作用最明显。茜素红染色显示,10-7mol/L辛伐他汀能促进细胞矿化结节形成;RT-PCR结果表明,辛伐他汀促进细胞碱性磷酸酶和骨形成蛋白-2基因表达呈时间依赖性增高。结论:10-7mol/L的辛伐他汀不仅有利于人牙周膜成纤维细胞增殖,而且促进其成骨分化和相关基因的表达。

Objective： To investigate the effects of simvastatin on the viability and osteoblastic differentiation of human periodontal ligament fibroblast （PDLFs） in vivo. Method： Human PDLFs were obtained from premolars and cultured in vivo. Human PDLFs of passage four were incubated with simvastatin at various concentrations：0, 10-8, 10-7, 10-6, 10-5 and 10-4 mol/L, and their viability and alkaline phosphatase （ALP） activity were measured. The osteoblastic differentiation were characterized by Alizarin Red-s （ARS） staining for calcium deposition. The levels of ALP and bone morphogenetic protein-2 （BMP-2） mRNA were evaluated by RT-PCR. Result： Simvastatin at 10-7 mol / L resulted in higher viability at 2 and 3 days of culture （P 〈 0.05）. The ALP activity of human PDLFs was improved in 10-s, 10-7, 10-6 and 10-5 mol / L simvastatin-treated groups （P 〈 0.05） and the peak level was attained at a concentration of 10-7 mol / L. After adminstration of 10-7 mol / L sim vastatin,the mineralized nodule formation was enhanced, and the expression of ALP and BMP-2 mRNA in human PDLFs were up-regulated in a time-dependent manner. Conclusion： A simvastatin treatment at optimal concentration can pose a positive effect on the metabolism of human PDLFs. Simvastatin stimulate the osteoblastic differentiation of human PDLFs and may be useful for regenerating periodontal hard tissue.