Gaultier Frédérick, Ejeil Anne-Laure, Dridi Sophie-Myriam, Piccardi Nathalie, Piccirilli Antoine, Msika Philippe, Pellat Bernard, Godeau Gaston, Gogly Bruno
Laboratory of Physiopathology of Non-Mineralized Tissues, U.F.R. Odontology, René Descartes University Paris V, Montrouge, France.
J Periodontol. 2005 Aug;76(8):1329-38. doi: 10.1902/jop.2005.76.8.1329.
In this study we examine the properties of a vegetable extract from seeds of Lupinus albus (LU 105). In previous works we demonstrated that LU 105 reduced the expression, by gingival fibroblasts, of both matrix metalloproteinase (MMP)-2 and MMP-9. We decided to study the impact of LU 105 on cell proliferation and morphology. Using organ culture media we also studied the MMP and tissue inhibitors of metalloproteinases (timp) expression AND THE cytokines secretion.
Healthy and inflamed gingival biopsies were placed in appendage culture with or without LU 105. The organ culture media were analyzed using Western blottings (MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, MMP-13, TIMP-1, and TIMP-2) and gelatine zymography. A reverse transcription polymerase chain reaction (RT-PCR) was also performed on healthy and inflamed gingival biopsies, which were maintained in culture with or without LU 105 0.1%. Then, we decided to determine the amount of cytokines present in the organ culture media such as interleukin (IL)-1 beta, IL-4, IL-6, transforming growth factor (TGF)-beta, and tumor necrosis factor (TNF)-alpha.
When gingival biopsies derived from inflamed tissues were cultured with LU 105 0.1% in the culture media, the MMP and TIMP expression and activity decreased significantly when compared to cultures without LU 105. Moreover, we did not note any statistical difference in the cell proliferation compared with human gingival fibroblast cultures without LU 105. Furthermore, IL-1 beta, IL-6, TGF-beta, and TNF-alpha amounts in the culture media decreased significantly, whereas IL-4 increased significantly when LU 105 0.1% was added to the culture media.
LU 105, a novel metalloproteinase inhibitor with few consequences on cell proliferation and morphology, is a vegetable extract with potential clinical capacity.
在本研究中,我们检测了白羽扇豆(LU 105)种子的植物提取物的特性。在之前的研究中,我们证明LU 105可降低牙龈成纤维细胞中基质金属蛋白酶(MMP)-2和MMP-9的表达。我们决定研究LU 105对细胞增殖和形态的影响。使用器官培养基,我们还研究了MMP和金属蛋白酶组织抑制剂(TIMP)的表达以及细胞因子的分泌。
将健康和发炎的牙龈活检组织置于含或不含LU 105的附属器培养中。使用蛋白质免疫印迹法(检测MMP-1、MMP-2、MMP-3、MMP-7、MMP-9、MMP-13、TIMP-1和TIMP-2)和明胶酶谱法分析器官培养基。对在含或不含0.1% LU 105的培养基中培养的健康和发炎的牙龈活检组织进行逆转录聚合酶链反应(RT-PCR)。然后,我们决定测定器官培养基中存在的细胞因子的量,如白细胞介素(IL)-1β、IL-4、IL-6、转化生长因子(TGF)-β和肿瘤坏死因子(TNF)-α。
当来自发炎组织的牙龈活检组织在培养基中与0.1% LU 105一起培养时,与不含LU 105的培养相比,MMP和TIMP的表达及活性显著降低。此外,与不含LU 105的人牙龈成纤维细胞培养相比,我们未发现细胞增殖有任何统计学差异。此外,当向培养基中添加0.1% LU 105时,培养基中IL-1β、IL-6、TGF-β和TNF-α的量显著降低,而IL-4显著增加。
LU 105是一种对细胞增殖和形态影响较小的新型金属蛋白酶抑制剂,是一种具有潜在临床应用能力的植物提取物。