CAJ | 학술논문

目的：观察牙髓卟啉单胞菌超声提取物对成骨细胞凋亡的促进作用，并初步探讨其相关分子机制。方法：用浓度为1、10、100μg/mL牙髓卟啉单胞菌超声提取物作用于成骨细胞，分别采用Hoechst33258染色法观察细胞的形态变化；比色法检测细胞中的Caspase－3酶活性改变；同时通过Caspase抑制剂阻断实验分析相关成骨细胞的凋亡途径。结果：牙髓卟啉单胞菌超声提取物能使成骨细胞形态出现染色质浓缩、碎裂等典型的凋亡改变，并可显著提高成骨细胞的Caspase－3酶活性，且具有浓度依赖性（P＜0．05）；Caspase－8抑制剂和Caspase－9抑制剂可以部分抑制牙髓卟啉单胞菌提取物对成骨细胞的促凋亡作用，而Caspase－3抑制剂则能完全阻断牙髓卟啉单胞菌对成骨细胞的促凋亡作用。结论：牙髓卟啉单胞菌超声提取物具有促成骨细胞凋亡的作用，细胞内、外凋亡途径都参与了该过程，并且是一种依赖Caspase－3的凋亡通路。
목적：관찰아수계람단포균초성제취물대성골세포조망적촉진작용，병초보탐토기상관분자궤제。방법：용농도위1、10、100μg/mL아수계람단포균초성제취물작용우성골세포，분별채용Hoechst33258염색법관찰세포적형태변화；비색법검측세포중적Caspase－3매활성개변；동시통과Caspase억제제조단실험분석상관성골세포적조망도경。결과：아수계람단포균초성제취물능사성골세포형태출현염색질농축、쇄렬등전형적조망개변，병가현저제고성골세포적Caspase－3매활성，차구유농도의뢰성（P＜0．05）；Caspase－8억제제화Caspase－9억제제가이부분억제아수계람단포균제취물대성골세포적촉조망작용，이Caspase－3억제제칙능완전조단아수계람단포균대성골세포적촉조망작용。결론：아수계람단포균초성제취물구유촉성골세포조망적작용，세포내、외조망도경도삼여료해과정，병차시일충의뢰Caspase－3적조망통로。
AIM:To investigate the apoptosis of human osteoblastic hFOB 1.19 cells induced by sonicated extract of P.endodontalis and the relevant molecular mechanism.METHODS:After exposure to sonicated extract of P.endodontalis at 0 (the control),1,10 and 100 μg/mL for 48 h,the morphology changes of hFOB 1.19 cells were observed by Hoechst33258 staining.Caspase-3 activity was detected by colorimetric assay .The apoptosis rate was an-alyzed by flow cytometry after the cells had been treated with the extract of P.endodontalis plus Caspase inhibitors.RE-SULTS:The sonicated extract of P.endodontalis induced chromatin concentration,breakup and other typical apoptosis changes in hFOB 1.19 cells.Meanwhile,the extract increased Caspase-3 activity of hFOB 1.19 cells in a dose-de-pendent manner(P<0.05)Caspase-8 inhibitor and Caspase-9 inhibitor partially inhibited the apoptosis of hFOB 1. 19 cells induced by the extract.Caspase-3 inhibitor completely blocked the apoptosis of hFOB 1.19 cells.CONCLU-SION:The extract of P.endodontalis can promote hFOB 1.19 cell apoptosis.Caspase-3-dependent apoptosis pathway is involved in this process,including both internal and external pathway.