Vascular endothelial growth factor is regulated by the canonical and non-canonical transforming growth factor-β pathway in synovial fibroblasts derived from osteoarthritis patients
Vascular endothelial growth factor is regulated by the canonical and non-canonical transforming growth factor-β pathway in synovial fibroblasts derived from osteoarthritis patients
复制标题
骨关节炎患者滑膜成纤维细胞中血管内皮生长因子受经典和非经典转化生长因子-β途径的调节
DOI：
10.1155/2019/6959056
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Takaso M.
中科院分区：
文献类型：
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作者：
Takano S;Uchida K;Shoji S;Itakura M;Iwase D;Aikawa J;Mukai M;Sekiguchi H;Inoue G;Takaso M.
背景资料。先前的研究表明，血管内皮生长因子(VEGF)与膝骨性关节炎(OA)患者的严重程度和疼痛有关。转化生长因子-β(TGFR-β)可诱导滑膜成纤维细胞表达血管内皮生长因子。然而，转化生长因子β介导的血管内皮细胞生长因子调控的信号通路尚未明确。对接受全膝关节置换术的骨性关节炎患者的滑膜组织(SYT)进行提取，并对滑膜组织进行培养。用培养基组(对照组)、人重组转化生长因子β(hrTGFβ)、hrTGFTAK5抑制剂SB505124、hrTGFTAK1抑制剂(5Z)-7-oxozeaenol或hrTGFTAK1 p38抑制剂SB203580刺激细胞6 h，用实时定量聚合酶链式反应检测细胞中VEGFmRNA的表达，用酶联免疫吸附试验检测细胞上清液中VEGFmRNA的表达。此外，用免疫印迹法检测Smad2和p38的磷酸化水平。Alk5(SB505124)和TAK1(5Z-oxozeaenol)抑制剂可完全抑制转化生长因子β诱导的VEGFmRNA表达和蛋白表达。SB505124和5Z-oxozeaenol也抑制Smad2和p38的磷酸化。P38抑制剂(SB203580)可部分抑制转化生长因子β介导的VEGFmRNA和蛋白的产生。转化生长因子β对骨性关节炎患者系统性红斑狼疮中血管内皮生长因子表达和血管内皮生长因子蛋白产生的调节通过规范和非规范两种途径进行。
Background. Previous studies suggest the presence of an association of vascular endothelial growth factor (VEGF) with osteoarthritis (OA) severity and pain in patients with knee OA. VEGF expression in human synovial fibroblasts (SFs) is induced by transforming growth factor‐beta (TGFβ). However, the signaling pathway governing TGFβ‐mediated regulation of VEGF in SFs has not been identified.Methods. OA patients who underwent total knee arthroplasty had their synovial tissue (SYT) extracted and the constituent SFs cultured. The cells were stimulated with culture medium (control), human recombinant TGFβ(hrTGFβ), hrTGFβ+ ALK5 inhibitor SB505124, hrTGFβ+ transforming growth factor activating kinase 1 (TAK1) inhibitor (5Z)‐7‐oxozeaenol, or hrTGFβ+ p38 inhibitor SB203580 for 6 h.VEGFmRNA expression in SFs was examined using real‐time polymerase chain reaction and VEGF protein production in the cell supernatant was examined using enzyme‐linked immunosorbent assay. Additionally, phosphorylated levels of SMAD2 and p38 were examined using western blotting.Results. ALK5 (SB505124) and TAK1 (5Z‐oxozeaenol) inhibitors completely suppressed TGFβ‐inducedVEGFmRNA expression and VEGF protein production. Both SB505124 and 5Z‐oxozeaenol also suppressed SMAD2 and p38 phosphorylation. The p38 inhibitor (SB203580) partially inhibited TGFβ‐mediatedVEGFmRNA and VEGF protein production.Conclusion. TGFβ‐mediated regulation ofVEGFexpression and VEGF protein production in the SYT of OA patients occurs through both the canonical and noncanonical pathway.