Kuo Po-Jan, Lin Hsiao-Lun, Lin Chi-Yu, Chin Yu-Tang, Tu Hsiao-Pei, Lai Tat-Ming, Chiu Hsien-Chung, Fu Earl
Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China.
Department of Periodontology, National Defense Medical Center and Tri-Service General Hospital, Taipei, Taiwan, Republic of China.
J Periodontol. 2016 Oct;87(10):1228-37. doi: 10.1902/jop.2016.140653. Epub 2016 Jun 13.
Matrix metalloproteinases (MMPs) play a key role in inflammatory periodontal disease. Synergistically enhanced MMP-2 expression in a coculture of human gingival fibroblasts (HGFs) and human monocytic U937 cells was observed. Crosstalk between these two cells via the extracellular matrix metalloproteinase inducer (EMMPRIN) was demonstrated.
Enzyme levels of MMP-2 in HGFs and direct coculture with U937 were examined by zymography. MMP-2 and EMMPRIN expressions of HGFs and U937 were determined in coculture and conditioned cultures (using supernatants from HGF- or U937-conditioned medium). The crosstalk was evaluated by EMMPRIN extrasupplement and EMMPRIN inhibition, through pretreatment of U937 with cyclosporine-A.
Direct coculturing of HGFs and U937 enhanced MMP-2 enzyme level and mRNA expression. Coculturing also increased membranous EMMPRIN expression of U937, but not from HGFs. In conditioned cultures, mRNA expression of MMP-2 increased in HGFs which received U937-conditioned medium. Increased MMP-2 was not observed in U937 with HGF-conditioned medium, although mRNA expression of EMMPRIN increased. Enhanced MMP-2 was observed after administration of exogenous EMMPRIN in HGFs; however, reduced MMP-2 enzyme level was noted if EMMPRIN of cocultured U937 was inhibited.
In the coculture of HGFs and U937, upregulated EMMPRIN expression in U937, which may be triggered by HGFs, can enhance MMP-2 expression in HGFs. Crosstalk between HGFs and U937 involving MMP-2 from HGFs was proposed; EMMPRIN from U937 may play a particular role.
基质金属蛋白酶（MMPs）在炎症性牙周病中起关键作用。在人牙龈成纤维细胞（HGFs）和人单核细胞U937细胞的共培养中观察到MMP-2表达协同增强。已证实这两种细胞通过细胞外基质金属蛋白酶诱导剂（EMMPRIN）发生相互作用。
通过酶谱法检测HGFs中MMP-2的酶水平以及与U937的直接共培养情况。在共培养和条件培养（使用HGF或U937条件培养基的上清液）中测定HGFs和U937的MMP-2和EMMPRIN表达。通过用环孢素A预处理U937，经EMMPRIN额外补充和EMMPRIN抑制来评估这种相互作用。
HGFs与U937的直接共培养增强了MMP-2酶水平和mRNA表达。共培养还增加了U937的膜性EMMPRIN表达，但HGFs未增加。在条件培养中，接受U937条件培养基的HGFs中MMP-2的mRNA表达增加。在使用HGF条件培养基的U937中未观察到MMP-2增加，尽管EMMPRIN的mRNA表达增加。在HGFs中给予外源性EMMPRIN后观察到MMP-2增强；然而，如果共培养的U937的EMMPRIN被抑制，则MMP-2酶水平降低。
在HGFs和U937的共培养中，U937中上调的EMMPRIN表达（可能由HGFs触发）可增强HGFs中MMP-2的表达。提出了HGFs和U937之间涉及HGFs的MMP-2的相互作用；U937的EMMPRIN可能起特定作用。
