Sulfur dioxide inhibits vascular smooth muscle cell proliferation via suppressing the Erk/MAP kinase pathway mediated by cAMP/PKA signaling.
Sulfur dioxide inhibits vascular smooth muscle cell proliferation via suppressing the Erk/MAP kinase pathway mediated by cAMP/PKA signaling.
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二氧化硫通过抑制 cAMP/PKA 信号传导介导的 Erk/MAP 激酶途径抑制血管平滑肌细胞增殖
DOI：
10.1038/cddis.2014.229
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发表时间：
2014-05-22
影响因子：
9
通讯作者：
中科院分区：
文献类型：
--
作者：
本研究旨在探讨内源性二氧化硫（SO2）对血管平滑肌细胞（VSMC）增殖的影响，以及环磷酸腺苷（cAMP）/蛋白激酶A（PKA）和细胞外信号调节激酶（Erk）/丝裂原活化蛋白激酶（MAPK）通路之间的相互作用。通过细胞计数、生长曲线、流式细胞术和BrdU标记等方法，发现SO2通过阻止细胞周期由G1期进入S期和减少DNA合成而抑制VSMC增殖。Western blot分析显示，SO2合酶天冬氨酸氨基转移酶（AAT 1和AAT 2）过表达显著抑制血清诱导的VSMCs增殖细胞核抗原（PCNA）蛋白表达。此外，AAT 1或AAT 2的过表达显着减少了血清处理的VSMC中BrdU的掺入。相比之下，AAT 1或AAT 2敲低显著加剧了血清刺激的VSMC增殖。因此，外源性和内源性SO2抑制血清诱导的VSMC增殖。Annexin V-碘化丙啶（PI）染色和细胞周期分析表明，SO2对血清诱导的VSMC增殖模型中的细胞凋亡无影响。在血小板衍生生长因子（PDGF）BB刺激的VSMC增殖模型中，SO2使PDGF BB诱导的Erk 1/2、MAPK激酶1/2和RAF原癌基因丝氨酸/苏氨酸蛋白激酶（c-Raf）活性位点脱磷酸化。然而，Erk/MAPK途径中的三种激酶的失活不是由于SO2同时单独干扰它们，而是由于SO2影响了c-Raf分子的上游活性。因此，我们研究了cAMP/PKA通路，其可以抑制VSMCs中的Erk/MAPK转导。结果表明，SO2可激活cAMP/PKA通路，阻断c-Raf的激活，而c-Raf上的Ser 259位点在SO2抑制Erk/MAPK通路中起重要作用。本研究首次证实SO2通过抑制cAMP/PKA信号通路介导的Erk/MAPK信号通路对VSMC增殖具有负性调节作用。
The present study was designed to investigate the role of endogenous sulfur dioxide (SO 2) in vascular smooth muscle cell (VSMC) proliferation, and explore the possible role of cross-talk between cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) and extracellular signal-regulated kinase (Erk)/mitogen-activated protein kinase (MAPK) pathways in this action. By cell counting, growth curve depict, flow cytometry and bromodeoxyuridine (BrdU) labeling assays, we found that SO 2 inhibited VSMC proliferation by preventing cell cycle progression from G1 to S phase and by reducing DNA synthesis. SO 2 synthase aspartate aminotransferase (AAT1 and AAT2) overexpression significantly inhibited serum-induced proliferating cell nuclear antigen (PCNA) protein expression in VSMCs, demonstrated by western blot analysis. Moreover, overexpression of AAT1 or AAT2 markedly reduced incorporation of BrdU in serum-treated VSMCs. By contrast, either AAT1 or AAT2 knockdown significantly exacerbated serum-stimulated VSMC proliferation. Thus, both exogenous-and endogenous-derived SO 2 suppressed serum-induced VSMC proliferation. However, annexin V-propidium iodide (PI) staining and cell cycle analysis demonstrated that SO 2 did not influence VSMC apoptosis in the serum-induced proliferation model. In a platelet-derived growth factor (PDGF)-BB-stimulated VSMC proliferation model, SO 2 dephosphorylated the active sites of Erk1/2, MAPK kinase 1/2 and RAF proto-oncogene serine/threonine-protein kinase (c-Raf) induced by PDGF-BB. However, the inactivation of the three kinases of the Erk/MAPK pathway was not due to the separate interferences on them by SO 2 simultaneously, but a consequence of the influence on the upstream activity of the c-Raf molecule. Hence, we examined the cAMP/PKA pathway, which could inhibit Erk/MAPK transduction in VSMCs. The results showed that SO 2 could stimulate the cAMP/PKA pathway to block c-Raf activation, whereas the Ser259 site on c-Raf had an important role in SO 2-induced suppression of Erk/MAPK pathway. The present study firstly demonstrated that SO 2 exerted a negative regulation of VSMC proliferation via suppressing the Erk/MAPK pathway mediated by cAMP/PKA signaling.
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影响因子：
8
作者：
Kawada, M;Yamagoe, S;Uehara, Y
通讯作者：
Uehara, Y
DOI：
10.1073/pnas.90.21.10300
发表时间：
1993-11-01
影响因子：
11.1
作者：
GRAVES, LM;BORNFELDT, KE;KREBS, EG
通讯作者：
KREBS, EG
影响因子：
5.3
作者：
Dumaz, N;Light, Y;Marais, R
通讯作者：
Marais, R
影响因子：
5.3
作者：
Light, Y;Paterson, H;Marais, R
通讯作者：
Marais, R
DOI：
10.1073/pnas.70.6.1753
发表时间：
1973-01-01
影响因子：
11.1
作者：
BENDITT, EP;BENDITT, JM
通讯作者：
BENDITT, JM