The role of mitogen-activated protein kinase and protein kinase C in fibronectin production in human vascular smooth muscle cells.
The role of mitogen-activated protein kinase and protein kinase C in fibronectin production in human vascular smooth muscle cells.
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丝裂原激活蛋白激酶和蛋白激酶 C 在人血管平滑肌细胞纤连蛋白产生中的作用。
DOI：
10.1006/jsre.1999.5646
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Kent,KC
中科院分区：
文献类型：
--
作者：
Kaiura,TL;Itoh,H;Kent,KC
背景在评估各种生长因子、细胞因子和细胞外基质（ECM）蛋白后，我们发现平滑肌细胞（SMC）纤连蛋白（Fn）产生的最有效激动剂是转化生长因子-β（TGF-β）和表皮生长因子（EGF）。为了确定参与该基质蛋白产生的可能信号通路，我们研究了细胞内蛋白、蛋白激酶 C (PKC) 和丝裂原激活蛋白激酶 (MAP-K) 在 TGF-β 和 EGF 诱导的人血管 SMC Fn 产生中的作用。材料和方法用 TGF-β (10 ng/ml) 和 EGF (100 ng/ml) 刺激人 SMC 后，测定细胞培养基中的 Fn通过使用特定抗体进行免疫印迹。 PKC 通过佛波醇 12,13-二丁酸酯 (PDBu) 短暂刺激 SMC 来激活，并通过 PDBu 或抑制剂 GF109203X 的下调来抑制。 PD098059 抑制 MAP-K。结果 PKC 激活增加基础并协同增强 TGF-β 和 EGF 诱导的 Fn 产生。然而，通过下调和 GF109203X 抑制 PKC 并不会减少 TGF-β 和 EGF 产生的 Fn。令人惊讶的是，这两种抑制方法略微增加了基础和激动剂诱导的 Fn 产生。 MAP-K 激酶抑制剂 PD098059 几乎完全抑制 EGF 并部分抑制 TGF-β 诱导的 Fn 产生。结论 PKC 的激活刺激 Fn 产生；然而，TGF-β 和 EGF 都不通过 PKC 依赖性途径产生 Fn。 EGF 和 TGF-β 都至少部分通过细胞内信号蛋白 MAP-K 刺激 Fn 产生。了解细胞外基质蛋白产生所涉及的信号通路将有助于设计内膜增生的特异性抑制剂。
BackgroundAfter evaluating various growth factors, cytokines, and extracellular matrix (ECM) proteins, we found that the most potent agonists of smooth muscle cell (SMC) fibronectin (Fn) production were transforming growth factor-beta (TGF-β) and epidermal growth factor (EGF). To determine the possible signaling pathways involved in the production of this matrix protein, we investigated the role of the intracellular proteins, protein kinase C (PKC) and mitogen-activated protein kinase (MAP-K), in TGF-β- and EGF-induced human vascular SMC Fn production.Materials and MethodsAfter stimulation of human SMCs with TGF-β (10 ng/ml) and EGF (100 ng/ml), Fn in the cell medium was assayed by immunoblotting using a specific antibody. PKC was activated by brief stimulation of SMC with phorbol 12,13-dibutyrate (PDBu) and inhibited by downregulation with PDBu or the inhibitor, GF109203X. MAP-K was inhibited with PD098059.ResultsPKC activation increased basal and synergistically enhanced TGF-β- and EGF-induced Fn production. However, inhibition of PKC by downregulation and GF109203X did not diminish Fn production by TGF-β and EGF. Surprisingly, these two methods of inhibition slightly increased basal and agonist-induced Fn production. The MAP-K kinase inhibitor, PD098059, produced an almost complete inhibition of EGF and a partial inhibition of TGF-β-induced Fn production.ConclusionsActivation of PKC stimulates Fn production; however, neither TGF-β nor EGF produce Fn through a PKC-dependent pathway. EGF and TGF-β both stimulate Fn production at least in part through the intracellular signaling protein MAP-K. Understanding the signaling pathways involved in extracellular matrix protein production will allow the design of specific inhibitors of intimal hyperplasia.