The activation of p38 MAPK limits the abnormal proliferation of vascular smooth muscle cells induced by high sodium concentrations.
The activation of p38 MAPK limits the abnormal proliferation of vascular smooth muscle cells induced by high sodium concentrations.
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p38 MAPK的激活限制了高钠浓度诱导的血管平滑肌细胞的异常增殖
DOI：
10.3892/ijmm.2015.2394
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发表时间：
2016-01
影响因子：
5.4
通讯作者：
Yuan Z
中科院分区：
文献类型：
--
作者：
Wu Y;Zhou J;Wang H;Wu Y;Gao Q;Wang L;Zhao Q;Liu P;Gao S;Wen W;Zhang W;Liu Y;Yuan Z
本研究的目的是确定高钠水平是否可以直接促进血管平滑肌细胞（VSMC）的增殖并阐明其潜在机制。向常规培养基中添加额外的氯化钠 (NaCl)。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑 (MTT) 测定和 5-乙炔基-2'-脱氧尿苷 (EdU) 掺入测定评估细胞增殖。通过逆转录定量聚合酶链反应（RT-qPCR）测量增殖细胞核抗原（PCNA）的mRNA表达水平。通过蛋白质印迹分析测量 PCNA 和磷酸化 c-Jun 氨基 N 末端激酶 (p-JNK)、磷酸化细胞外信号调节激酶 1/2 (p-ERK1/2) 和磷酸化 p38 丝裂原激活蛋白激酶 (p-p38 MAPK) 的蛋白表达水平。细胞增殖实验显示Na+而非Cl-或渗透压促进VSMC的增殖。高钠水平上调 PCNA 的表达以及 JNK、ERK1/2 和 p38 MAPK 的磷酸化水平。 JNK 和 ERK1/2 的抑制降低了 PCNA 的表达。值得注意的是，使用抑制剂 SB203580 抑制 p38 MAPK 会增加 PCNA 表达。然而，当p38 MAPK被茴香霉素激活时，PCNA表达降低。总的来说，我们的研究结果表明，相对较高的钠水平本身可以通过 JNK/ERK1/2/PCNA 途径直接促进 VSMC 的增殖。同时，高钠水平引起的 VSMC 增殖诱导可以通过 p38 MAPK 的激活维持在较低水平。
The aim of the present study was to ascertain whether high sodium levels can directly promote the proliferation of vascular smooth muscle cells (VSMCs) and to elucidate the underlying mechanisms. Additional sodium chloride (NaCl) was added to the routine culture medium. Cell proliferation was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay. The mRNA expression level of proliferating cell nuclear antigen (PCNA) was measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The protein expression levels of PCNA and phosphorylated c-Jun amino N-terminal kinase (p-JNK), phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) and phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) were measured by western blot analysis. Cell proliferation assay revealed that Na+ rather than Cl− or osmotic pressure promoted the proliferation of the VSMCs. The high sodium level upregulated the expression of PCNA and the phosphorylation levels of JNK, ERK1/2 and p38 MAPK. The inhibition of JNK and ERK1/2 decreased PCNA expression. Of note, the inhibition of p38 MAPK using the inhibitor, SB203580, increased PCNA expression. However, when p38 MAPK was activated by anisomycin, PCNA expression was decreased. On the whole, our findings demonstrate that a relatively high sodium level per se directly promotes the proliferation of VSMCs through the JNK/ERK1/2/PCNA pathway. At the same time, this induction of the proliferation of VSMCs due to high sodium levels can be maintained at a low level via the activation of p38 MAPK.