NOX4 mediates BMP4-induced upregulation of TRPC1 and 6 protein expressions in distal pulmonary arterial smooth muscle cells.
NOX4 mediates BMP4-induced upregulation of TRPC1 and 6 protein expressions in distal pulmonary arterial smooth muscle cells.
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NOX4 介导 BMP4 诱导的远端肺动脉平滑肌细胞中 TRPC1 和 6 蛋白表达的上调
DOI：
10.1371/journal.pone.0107135
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Jiang Q;Fu X;Tian L;Chen Y;Yang K;Chen X;Zhang J;Lu W;Wang J
基本原理我们之前的研究表明，骨形态发生蛋白 4 (BMP4) 介导的经典瞬时受体电位 (TRPC) 表达升高在很大程度上解释了肺动脉平滑肌细胞 (PASMC) 增殖的增强。在本研究中，我们试图确定 BMP4 上调 TRPC 表达的信号通路。方法我们采用重组人 BMP4 (rhBMP4) 来确定 BMP4 对大鼠远端 PASMC 中 NADPH 氧化酶 4 (NOX4) 和活性氧 (ROS) 产生的影响。我们还设计了靶向NOX4的小干扰RNA（siNOX4），并检测NOX4敲低是否影响rhBMP4诱导的PASMCs中ROS、TRPC1和6的表达、细胞增殖和细胞内Ca2+测定。结果rhBMP4处理的大鼠远端PASMC中，NOX4表达为（226.73±11.13）%，平均ROS水平为未处理对照细胞的（123.65±1.62）%。 siNOX4 转染显着降低了 rhBMP4 诱导的 PASMC 中平均 ROS 水平的升高。此外，siNOX4 转染显着降低了 rhBMP4 诱导的 TRPC1 和 6 蛋白、基础 [Ca2+]i 和 SOCE 的升高。此外，与对照组（0.21±0.001）相比，rhBMP4处理的细胞增殖显着增强（0.41±0.001）（P<0.01）。然而，这种增加因 NOX4 的敲低而减弱。此外，外部ROS（H2O2 100 µM，24小时）挽救了NOX4敲低的影响，包括TRPC1和6表达、基础细胞内钙浓度（[Ca2+]i）和钙库操作钙内流（SOCE）的下降，表明NOX4在BMP4诱导的增殖和细胞内钙稳态中发挥着重要的调节作用。结论这些结果表明BMP4可能通过上调PASMCs中NOX4的表达来增加ROS水平，增强TRPC1和6的表达和增殖。
Rationale Our previous studies demonstrated that bone morphogenetic protein 4 (BMP4) mediated, elevated expression of canonical transient receptor potential (TRPC) largely accounts for the enhanced proliferation in pulmonary arterial smooth muscle cells (PASMCs). In the present study, we sought to determine the signaling pathway through which BMP4 up-regulates TRPC expression. Methods We employed recombinant human BMP4 (rhBMP4) to determine the effects of BMP4 on NADPH oxidase 4 (NOX4) and reactive oxygen species (ROS) production in rat distal PASMCs. We also designed small interfering RNA targeting NOX4 (siNOX4) and detected whether NOX4 knockdown affects rhBMP4-induced ROS, TRPC1 and 6 expression, cell proliferation and intracellular Ca2+ determination in PASMCs. Results In rhBMP4 treated rat distal PASMCs, NOX4 expression was (226.73±11.13) %, and the mean ROS level was (123.65±1.62) % of that in untreated control cell. siNOX4 transfection significantly reduced rhBMP4-induced elevation of the mean ROS level in PASMCs. Moreover, siNOX4 transfection markedly reduced rhBMP4-induced elevation of TRPC1 and 6 proteins, basal [Ca2+]i and SOCE. Furthermore, compared with control group (0.21±0.001), the proliferation of rhBMP4 treated cells was significantly enhanced (0.41±0.001) (P<0.01). However, such increase was attenuated by knockdown of NOX4. Moreover, external ROS (H2O2 100 µM, 24 h) rescued the effects of NOX4 knockdown, which included the declining of TRPC1 and 6 expression, basal intracellular calcium concentration ([Ca2+]i) and store-operated calcium entry (SOCE), suggesting that NOX4 plays as an important mediator in BMP4-induced proliferation and intracellular calcium homeostasis. Conclusion These results suggest that BMP4 may increase ROS level, enhance TRPC1 and 6 expression and proliferation by up-regulating NOX4 expression in PASMCs.
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DOI：
10.1152/ajplung.00448.2004
发表时间：
2005-06-01
影响因子：
4.9
作者：
Wang, J;Shimoda, LA;Sylvester, JT
通讯作者：
Sylvester, JT
影响因子：
3.3
作者：
Diebold I;Petry A;Hess J;Görlach A
通讯作者：
Görlach A
影响因子：
20.1
作者：
Sorescu, GP;Song, H;Jo, H
通讯作者：
Jo, H
DOI：
10.1152/ajplung.00058.2008
发表时间：
2008-07-01
影响因子：
4.9
作者：
Lu, Wenju;Wang, Jian;Sylvester, J. T.
通讯作者：
Sylvester, J. T.
DOI：
10.1152/ajplung.00319.2003
发表时间：
2004-04-01
影响因子：
4.9
作者：
Wang, J;Shimoda, LA;Sylvester, JT
通讯作者：
Sylvester, JT