特定序列寡核苷酸MT01对牙龈卟啉单胞菌感染的成骨细胞的增殖、细胞周期及凋亡的影响

Effect of specific sequence oligodeoxynucleotide MT01 on the proliferation, apoptosis, and cell cycle of osteoblasts invaded by Porphyromonas gingivalis

目的通过观察MT01对感染状态下成骨细胞细胞增殖、细胞周期及凋亡的影响,探讨MT01对牙周致病菌感染的成骨细胞生物学性能的影响。方法以人成骨细胞MG63为目的细胞,分别与PBS、MT01、Cp G ODN、甲硝唑和庆大霉素共培养3 h后,按感染复数100∶1的比例加入牙周致病菌牙龈卟啉单胞菌共培养,检测培养24 h和48 h后MG63细胞的增殖、细胞周期及凋亡,以PBS作为空白对照,以牙龈卟啉单胞菌和PBS共培养作为阴性对照。结果 MT01+牙龈卟啉单胞菌组细胞增殖高于空白对照组(P<0.05),G1期细胞百分比低于阴性对照组,而S期细胞百分比高于阴性对照组(P<0.05),细胞早期凋亡率低于阴性对照组(P<0.05)。结论 MT01可促进感染状态下成骨细胞的细胞增殖,降低G1期细胞百分比,促使细胞进入S期并抑制细胞早期凋亡。

Objective This aimed to investigate the effect of specific sequence oligodeoxynucleotide MT01 on the biological properties of osteoblasts invaded by Porphyromonas gingivalis（P. gingivalis） by evaluating proliferation, cell cycle, and apoptosis. Methods MG63 osteoblasts were recovered and incubated with MT01, Cp G ODN, metronidazole（MNZ）, and gentamicin（GEN） for 3 h. P. gingivalis（the multiplicity of infection was 100∶1） was added subsequently and cocultured for another 24 and 48 h. Cells with PBS comprised the blank group, whereas cells with P. gingivalis comprised the negative controls. Six experimental groups were established： PBS group, P. gingivalis group, MT01＋P. gingivalis group, Cp G ODN＋P. gingivalis group, MNZ＋P. gingivalis group, and GEN＋P. gingivalis group. The proliferative ability was measured by methyl thiazolyl tetrazolium assay, and the percentages of apoptosis and cell cycle were examined by flow cytometry. Results Compared with the blank group, proliferation increased significantly in the MT01＋P. gingivalis group（P〈0.05）. The ratio of cells was lower at the G1 phase and higher at the S phase in the MT01＋P. gingivalis group compared with the results in the P. gingivalis group（P〈0.05）. Early cell apoptosis in the MT01＋P. gingivalis group was significantly lower than that in the P. gingivalis group（P〈0.05）. Conclusion MT01 can promote the proliferation, reduce the ratio of the G1 phase, increase the ratio of the S phase, and inhibit the early apoptosis of osteoblasts invaded by P. gingivalis.