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牙龈卟啉单胞菌对上皮细胞MMP - 9(IV型胶原酶)产生的影响。

Effect of Porphyromonas gingivalis on epithelial cell MMP-9 type IV collagenase production.

作者信息

Fravalo P, Ménard C, Bonnaure-Mallet M

机构信息

Equipe de Biologie Buccale, UFR Odontologie, Université de Rennes I, France.

出版信息

Infect Immun. 1996 Dec;64(12):4940-5. doi: 10.1128/iai.64.12.4940-4945.1996.

Abstract

Porphyromonas gingivalis is reportedly capable of stimulating the expression of host cell matrix metalloproteinases (MMP), contributing to tissue destruction. However, the impact of this bacterium on specific molecules remains to be determined. In this study, we evaluate the effect of P. gingivalis on regulation of MMP-9 expression in human gingival epithelial cells (HGEC). Various inocula of P. gingivalis were added to cultures of HGEC. The effects of live bacteria, heat-killed bacteria, and outer membrane extract were analyzed. MMP-9 secretion by HGEC was evaluated by enzyme-linked immunosorbent assay. For inocula smaller than one bacterium per cell, the quantity of MMP-9 secreted by HGEC was increased in comparison to control conditions. For inocula from 2.5 to 250 bacteria per cell, an inhibition of MMP-9 secretion in a dose-response fashion was observed, with a maximum reduction (ranging from 80 to 95% in five experiments) at 50 bacteria per cell. Gelatin zymograms confirmed the decrease in MMP-9 secretion. A band of 83 kDa, corresponding to activated enzyme, was present for inocula of 0.5 to 50 bacteria. Inhibition took place without any alteration of epithelial cell viability. Heat-killed bacteria and outer membrane extract also provoked proenzyme activation but did not inhibit MMP-9 secretion. These results demonstrate a direct effect of P. gingivalis on HGEC, suggesting a specific action on the collagen renewal process at the interface between the epithelium and connective tissue.

摘要

据报道,牙龈卟啉单胞菌能够刺激宿主细胞基质金属蛋白酶(MMP)的表达,从而导致组织破坏。然而,这种细菌对特定分子的影响仍有待确定。在本研究中,我们评估了牙龈卟啉单胞菌对人牙龈上皮细胞(HGEC)中MMP-9表达调控的影响。将不同接种量的牙龈卟啉单胞菌添加到HGEC培养物中。分析了活细菌、热灭活细菌和外膜提取物的作用。通过酶联免疫吸附测定法评估HGEC分泌的MMP-9。对于每个细胞接种量小于一个细菌的情况,与对照条件相比,HGEC分泌的MMP-9量增加。对于每个细胞接种2.5至250个细菌的情况,观察到MMP-9分泌呈剂量反应性抑制,在每个细胞接种50个细菌时抑制作用最大(在五个实验中降低幅度为80%至95%)。明胶酶谱证实了MMP-9分泌的减少。对于接种0.5至50个细菌的情况,存在一条对应于活化酶的83 kDa条带。抑制作用发生时上皮细胞活力没有任何改变。热灭活细菌和外膜提取物也能激活酶原,但不抑制MMP-9分泌。这些结果证明了牙龈卟啉单胞菌对HGEC有直接作用,表明其在上皮组织与结缔组织界面处的胶原更新过程中具有特定作用。

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