华西口腔医学杂志

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粪肠球菌对多形核白细胞释放基质金属蛋白酶-8及凋亡影响的实验研究

周梦宇 牛卫东   

  1. 大连医科大学口腔医学院口腔内科教研室, 辽宁大连116044
  • 收稿日期:2009-08-25 修回日期:2009-08-25 出版日期:2009-08-20 发布日期:2009-08-20
  • 通讯作者: 牛卫东,Tel:0411-86110399
  • 作者简介: 周梦宇(1977—),女,辽宁人,主治医师,硕士
  • 基金资助:

    国家自然科学基金资助项目(30870670)

Effect of Enterococcus faecalis on apoptosis rate and the release of matrix metalloproteinase-8 of polymorphonuclear leukocytes

ZHOU Meng-yu, NIU Wei-dong   

  1. Dept. of Oral Medicine, School of Stomatology, Dalian Medical University, Dalian 116044, China
  • Received:2009-08-25 Revised:2009-08-25 Online:2009-08-20 Published:2009-08-20
  • Contact: NIU Wei-dong,Tel:0411-86110399

摘要:

目的体外研究粪肠球菌对多形核白细胞(PMNs)释放基质金属蛋白酶-8(MMP-8)及凋亡的影响。方法提取PMNs,以加入粪肠球菌悬浮液的PMNs作为实验组;加入乙酸肉豆蔻佛波醇的PMNs为阳性对照组;PMNs的PBS悬浮液为阴性对照组。培养0、20、60、120 min后,ELISA法检测各时间点MMP-8的释放量。以加入粪肠球菌裂解液的PMNs为实验组,PMNs的PBS悬浮液为对照组,培养2、5、10、15 h后,流式细胞分析仪检测PMNs的凋亡率。结果在0 min时,实验组和阳性对照组MMP-8释放量间差异无统计学意义(P>0.01);在60、120 min时,实验组MMP-8的释放量明显低于阳性对照组,二者间差异有统计学意义(P<0.01)。在2、5、10、15 h,实验组凋亡率均高于对照组(P<0.01)。结论粪肠球菌作用于PMNs后,PMNs无大量释放MMP-8现象,PMNs无凋亡延迟现象。

关键词: 粪肠球菌, 多形核白细胞, 基质金属蛋白酶-8, 细胞凋亡

Abstract:

Objective To evaluate the release of matrix metalloproteinase -8(MMP-8) and apoptosis rate of polymorphonuclear leukocytes(PMNs) after PMNs was triggered by Enterococcus faecalis(E.faecalis) in vitro. Methods The activated E.faecalis suspension was prepared and added to PMNs suspension as experiment group. As a positive control, phorbol myristate acetate(PMA) was used. As negative control, PMNs suspension was incubated with PBS. The release of MMP-8 was measured at 0, 20, 60, 120 min by ELISA method. E.faecalis lysate acted on PMNs as experiment group, PMNs suspension was incubated with PBS as negative control, samples in two groups were incubated at 37 ℃ for 2, 5, 10, 15 h. The apoptosis rate of PMNs was tested by Flow Cytometry. Results At 0 min, there was no significant difference of MMP-8 release in the experiment group and positive control(P>0.01);whereas at 60, 120 min, E.faecalis induced a significant lower MMP -8 release compared with the positive control(P <0.01). The apoptosis rate of PMNs in both groups increased along with time, and apoptotic rate in experiment group was higher than that in the control group at 2, 5, 10, 15 h(P<0.01). Conclusion After E.faecalis act on PMNs,no significant release of MMP-8 from PMNs was observed. E.faecalis don′t induce PMNs apoptosis delay.

Key words: Enterococcus faecalis, polymorphonuclear leukocytes, matrix metalloproteinase-8, cell apoptosis