MT01对感染状态人成骨样细胞内ALP活性及mRNA表达的影响

Effects of MT01 on the ALP Activity and mRNA Expression of MG63 Infected by Porphyromonas Gingivalis

目的:通过检测MT01对牙龈卟啉单胞菌感染的人成骨样细胞内特异性成骨相关因子ALP活性及mRNA表达水平的改变,探讨MT01对感染状态下人成骨样细胞成骨向分化的影响。方法:选取状态良好的MG63细胞接种于6孔板内,2个MT01组加入质量浓度为1mg/L的MT01,共孵育3h后,相应组加入感染复数为100∶1的Pg菌悬液。实验分为:空白对照、MT01、Pg和MT01+Pg4组,碱性磷酸酶测试盒测定24h后上清液及细胞内ALP活性。Real-time PCR检测2、4、6、8、12、24h特异性成骨相关因子ALP mRNA的表达。结果:在感染与非感染情况下,MT01均可促进ALP活性,且上调MG63细胞内成骨相关因子ALP mRNA表达,其表达上调呈时间依赖性。结论:MT01可促进感染及非感染状态下MG63内ALP基因表达水平,提高ALP活力。

Objective：To study the effect of MT01 on the differentiation of osteoblasts infected by porphyromonas gingivalis（Pg）.Methods：The MG63 cells were seeded into 6-well cell culture plates.MT01 at a final concentration of 1μg·mL-1 was added and incubated for 3hours.Then the cells were challenged by Pg（MOI=100∶1）.There were four experimental groups：blank group,MT01,Pg and MT01＋Pg.ALP activities in the supernatant fluid and the cell lysate after 24 hours were detected by ALP kit.Real-time PCR was used to detect the mRNA expression of ALP after 2,4,6,8,12,and 24 hours.Results：MT01enhanced the ALP activity in infected or uninfected conditions.The mRNA expression of ALP in MG63 was upregulated by MT01post-infected by Pg or not in a time-dependent manner.Conclusion：MT01can upregulate the mRNA expression of ALP in Pg infected or uninfected MG63 and improve the ALP activity.