Role of PTPα in the destruction of periodontal connective tissues.
Role of PTPα in the destruction of periodontal connective tissues.
复制标题
DOI：
10.1371/journal.pone.0070659
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
McCulloch CA
中科院分区：
文献类型：
--
作者：
Rajshankar D;Sima C;Wang Q;Goldberg SR;Kazembe M;Wang Y;Glogauer M;Downey GP;McCulloch CA
IL-1β通过上调基质融解素-1 （MMP-3）参与结缔组织破坏，而基质融解素-1在成纤维细胞中是一个局灶黏着依赖性过程。蛋白酪氨酸磷酸酶-α (PTPα)富集并调节局灶粘连的形成，但PTPα在结缔组织破坏中的作用尚未明确。我们首先研究了结扎诱导的牙周炎对成年PTPα+/+和PTPα - / -小鼠牙周结缔组织的破坏，这增加了多种细胞因子的水平，包括IL-1β。结扎后3周，对上颌骨进行形态测量、显微计算机断层扫描和组织形态测量。与未结扎对照组相比，与PTPα - / -小鼠相比，WT的骨质流失量增加了1.5-3倍，牙龈固有层厚度减少了3倍，胶原纤维的数量减少了20倍。牙周组织免疫组化染色显示结扎部位MMP-3表达升高。其次，为了研究PTPα调节基质降解的机制，我们使用人MMP阵列筛选了经载体、IL-1β或TNFα处理的人牙龈成纤维细胞的条件培养基。虽然这两种细胞因子均可上调MMP-3，但只有IL-1β可刺激纤维连接蛋白修饰的人牙龈成纤维细胞的ERK活化。使用各种突变构建体或siRNA或PTPα ko以及匹配的野生型成纤维细胞，利用TIRF显微镜和免疫印迹分析，将PTPα活性缺失的细胞镀在纤维连接蛋白上，使其形成局灶粘连，并用IL-1β刺激。这些数据表明，PTPα在il -1β诱导的局灶黏附形成、ERK激活和MMP-3释放中具有催化和衔接功能。我们得出结论，炎症诱导的结缔组织降解涉及成纤维细胞，需要功能活跃的PTPα，部分是通过局灶粘连由IL-1β信号介导的。
IL-1β contributes to connective tissue destruction in part by up-regulating stromelysin-1 (MMP-3), which in fibroblasts is a focal adhesion-dependent process. Protein tyrosine phosphatase-α (PTPα) is enriched in and regulates the formation of focal adhesions, but the role of PTPα in connective tissue destruction is not defined. We first examined destruction of periodontal connective tissues in adult PTPα+/+ and PTPα−/− mice subjected to ligature-induced periodontitis, which increases the levels of multiple cytokines, including IL-1β. Three weeks after ligation, maxillae were processed for morphometry, micro-computed tomography and histomorphometry. Compared with unligated controls, there was ∼1.5–3 times greater bone loss as well as 3-fold reduction of the thickness of the gingival lamina propria and 20-fold reduction of the amount of collagen fibers in WT than PTPα−/− mice. Immunohistochemical staining of periodontal tissue showed elevated expression of MMP-3 at ligated sites. Second, to examine mechanisms by which PTPα may regulate matrix degradation, human MMP arrays were used to screen conditioned media from human gingival fibroblasts treated with vehicle, IL-1β or TNFα. Although MMP-3 was upregulated by both cytokines, only IL-1β stimulated ERK activation in human gingival fibroblasts plated on fibronectin. TIRF microscopy and immunoblotting analyses of cells depleted of PTPα activity with the use of various mutated constructs or with siRNA or PTPαKO and matched wild type fibroblasts were plated on fibronectin to enable focal adhesion formation and stimulated with IL-1β. These data showed that the catalytic and adaptor functions of PTPα were required for IL-1β-induced focal adhesion formation, ERK activation and MMP-3 release. We conclude that inflammation-induced connective tissue degradation involving fibroblasts requires functionally active PTPα and in part is mediated by IL-1β signaling through focal adhesions.
登录
查看更多内容
影响因子：
6.7
作者：
Astolfi, Claudia Maria;Shinohara, Andre Luis;de Souza, Ana Paula
通讯作者：
de Souza, Ana Paula
影响因子：
7.3
作者：
Lohinai, Z;Benedek, P;Szabó, C
通讯作者：
Szabó, C
影响因子：
4.3
作者：
Kubota, Takehiko;Itagaki, Manarni;Yoshie, Hiromasa
通讯作者：
Yoshie, Hiromasa
影响因子：
13.8
作者：
MAURO, LJ;DIXON, JE
通讯作者：
DIXON, JE
影响因子：
11.4
作者：
DENHERTOG, J;TRACY, S;HUNTER, T
通讯作者：
HUNTER, T