Aida Yukiko, Maeno Masao, Suzuki Naoto, Shiratsuchi Hisaka, Motohashi Masafumi, Matsumura Hideo
Department of Crown and Bridge Prosthodontics, Nihon University School of Dentistry, Tokyo 101-8310, Japan.
Life Sci. 2005 Nov 4;77(25):3210-21. doi: 10.1016/j.lfs.2005.05.052. Epub 2005 Jun 24.
Interleukin-1 (IL-1) plays key roles in altering cartilage matrix turnover. This turnover is regulated by matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs). In the present study, we examined the effect of IL-1beta on cell proliferation, alkaline phosphatase (ALPase) activity, and the expression of MMPs, and TIMPs in chondrocytes derived from normal human femoral cartilage. The cells were cultured in Dulbecco's modified Eagle's medium containing 15% fetal bovine serum and 0, 1, 10, or 100 U/ml of IL-1beta for up to 28 days. The level of expression of MMPs and TIMPs was estimated by determining mRNA levels using real-time PCR and by determining protein levels using an enzyme-linked immunosorbent assay. Cell proliferation decreased in the presence of IL-1beta after day 21 of culture. ALPase activity decreased significantly in the presence of IL-1beta after day 10 of culture. The expression of MMP-1, -2, and -3 increased markedly in the presence of IL-1beta after day 21 of culture. MMP-13 expression increased markedly in the presence of IL-1beta on day 1 of culture, but decreased markedly after day 7. The expression of TIMP-1 increased significantly after day 14 of culture. The expression of TIMP-2 decreased significantly on day 1, but increased significantly from day 3 to day 14 of culture. These results suggest that IL-1beta may stimulate cartilage matrix turnover by increasing mainly MMP-13 production by the cells.
白细胞介素-1（IL-1）在改变软骨基质周转方面发挥着关键作用。这种周转由基质金属蛋白酶（MMPs）和基质金属蛋白酶组织抑制剂（TIMPs）调节。在本研究中，我们检测了IL-1β对源自正常人股骨软骨的软骨细胞增殖、碱性磷酸酶（ALPase）活性以及MMPs和TIMPs表达的影响。将细胞培养在含有15%胎牛血清以及0、1、10或100 U/ml IL-1β的杜尔贝科改良伊格尔培养基中，培养长达28天。通过实时PCR测定mRNA水平以及通过酶联免疫吸附测定法测定蛋白质水平来评估MMPs和TIMPs的表达水平。培养21天后，在存在IL-1β的情况下细胞增殖下降。培养10天后，在存在IL-1β的情况下ALPase活性显著下降。培养21天后，在存在IL-1β的情况下MMP-1、-2和-3的表达显著增加。在培养第1天存在IL-1β时MMP-13表达显著增加，但在第7天后显著下降。培养14天后TIMP-1的表达显著增加。TIMP-2的表达在第1天显著下降，但在培养第3天至第14天显著增加。这些结果表明，IL-1β可能主要通过增加细胞产生MMP-13来刺激软骨基质周转。
