幽门螺杆菌空泡毒素对人牙周膜成纤维细胞的增殖和凋亡的影响

The effects of helicobacter pylori vacuolating cytotoxin on the proliferation and apoptosis of human periodontal ligament fibroblasts

目的:观察幽门螺杆菌空泡毒素(HP Vac A)对人牙周膜成纤维细胞(h PDLFs)增殖和凋亡的影响。方法:人牙周膜成纤维细胞株(h PDLFs100),分别与不同浓度的HP Vac A(0.1、0.2、0.3 mg/m L)共培养,CCK-8方法检测细胞的增殖情况;倒置显微镜观察细胞的形态变化;流式细胞仪检测细胞的凋亡情况;透射电镜观察细胞超微结构的改变。结果:随着HP Vac A浓度的增大和培养时间的延长,h PDLFs的增殖明显受到抑制;贴壁的PDLFs数量递减,细胞由长梭形逐渐变为圆形,折光性不断减弱,悬浮细胞和凋亡细胞增多;胞浆内大量空泡形成,细胞染色体聚集于核膜,呈块状、浓集和边缘化,表现出典型的凋亡特征。结论:HP Vac A对PDLFs有明显的细胞毒作用,能抑制细胞增殖,并引起细胞凋亡。

AIM： To observe the effects of helicobacte pylori vacuolating cytotoxin（ HP Vac A） on the proliferation and apoptosis of human periodontal ligament fibroblasts（ h PDLFs）. METHODS： h PDLFs were cultured in vitro with different concentrations（ 0. 1,0. 2,and 0. 3 mg / m L） of HP Vac A. Cell proliferation was examined by CCK- 8 method,the morphological changes were observed under inverted microscope,the apoptosis of the cells was detected by flow cytometry（ FCM）,and changes of cell ultrastructure was observed by transmission electron microscope.RESULTS： HP Vac A showed a concentration-and time-dependent inhibition on h PDLFs proliferation and stimulation on apoptosis. After treatment by HP Vac A the number of adherent h PDLFs decreased. The cell shape gradually changed from long fusiform into rounded and suspended. Apoptotic cells increased as detected by FCM. A large number of vacuoles within the cell cytoplasm was observed by transmission electron microscope,in which cell chromosomes were focus on karyotheca as massive,concentrated and marginalized presenting classical features of cell apoptosis.CONCLUTION： HP Vac A can inhibit the proliferation of periodontal ligament fibroblasts and cause cell apoptosis.