Yang Haihua, Long Feng, Zhang Youzhi, Yu Ronghuan, Zhang Peng, Li Wenjing, Li Shuijun, Jin Xianqiao, Xia Jingwen, Dong Liang, Zhu Ning, Huang Ying, Gong Yi, Chen Xiaodong
Department of Respiratory Medicine, Huashan Hospital North, Fudan University, ShangHai, China.
Department of Respiratory Medicine, Xuhui Central Hospital, ShangHai, China.
PLoS One. 2015 Apr 23;10(4):e0120515. doi: 10.1371/journal.pone.0120515. eCollection 2015.
Reduced neutrophil apoptosis plays an important role in the pathogenesis of acute exacerbation chronic obstructive pulmonary disease (AECOPD). The p38 mitogen-activated protein kinase (MAPK) signaling pathway is involved in neutrophil apoptosis. 1α,25-Dihydroxyvitamin D3 (1α,25VitD3) can induce tumor cell apoptosis. The aim of this study was to assess the effects of 1α,25VitD3 on peripheral blood neutrophil apoptosis in AECOPD and examine the role of the p38 MAPK signaling pathway.
The study enrolled 36 AECOPD patients and 36 healthy volunteers. Venous blood samples were obtained from both groups. Serum 25-hydroxyvitamin D (25-(OH) D) levels in peripheral venous blood were assayed using liquid chromatography-tandem mass spectrometry (LC-MS/MS); the neutrophils were separated and cultured with SB203580 (a p38 inhibitor) and 1α,25VitD3. Neutrophil apoptosis was measured using flow cytometry, and phospho-p38 MAPK protein expression was detected by Western blot. Statistical analysis was performed using analysis of variance. Student's t-test and Pearson's correlation coefficient were used for the between-group differences and correlation analysis, respectively.
The 25-(OH) D levels were lower in AECOPD patients than in healthy controls, and the peripheral blood neutrophil apoptosis results were similar. 1α,25VitD3 increased the apoptosis rate and the level of phospho-p38 MAPK in peripheral blood neutrophils of AECOPD patients. SB203580 partly inhibited 1α,25VitD3-induced peripheral blood neutrophil apoptosis and phospho-p38 MAPK overexpression. The 25-(OH) D levels were positively correlated with increased peripheral blood neutrophil apoptosis and phospho-p38 MAPK levels. In addition, expression of the phospho-p38 MAPK protein was also positively correlated with peripheral blood neutrophil apoptosis.
Our results suggest that 1α,25VitD3 induces peripheral blood neutrophil apoptosis through the p38 MAPK signaling pathway in AECOPD patients.
中性粒细胞凋亡减少在慢性阻塞性肺疾病急性加重期（AECOPD）的发病机制中起重要作用。p38丝裂原活化蛋白激酶（MAPK）信号通路参与中性粒细胞凋亡。1α,25-二羟维生素D3（1α,25VitD3）可诱导肿瘤细胞凋亡。本研究旨在评估1α,25VitD3对AECOPD患者外周血中性粒细胞凋亡的影响，并探讨p38 MAPK信号通路的作用。
本研究纳入36例AECOPD患者和36名健康志愿者。采集两组静脉血样本。采用液相色谱-串联质谱法（LC-MS/MS）检测外周静脉血中血清25-羟维生素D（25-(OH)D）水平；分离中性粒细胞，并用SB203580（一种p38抑制剂）和1α,25VitD3进行培养。采用流式细胞术检测中性粒细胞凋亡，通过蛋白质印迹法检测磷酸化p38 MAPK蛋白表达。采用方差分析进行统计分析。分别使用学生t检验和Pearson相关系数进行组间差异和相关性分析。
AECOPD患者的25-(OH)D水平低于健康对照组，外周血中性粒细胞凋亡结果相似。1α,25VitD3增加了AECOPD患者外周血中性粒细胞的凋亡率和磷酸化p38 MAPK水平。SB203580部分抑制了1α,25VitD3诱导的外周血中性粒细胞凋亡和磷酸化p38 MAPK的过表达。25-(OH)D水平与外周血中性粒细胞凋亡增加及磷酸化p38 MAPK水平呈正相关。此外，磷酸化p38 MAPK蛋白表达也与外周血中性粒细胞凋亡呈正相关。
我们的结果表明，1α,25VitD3通过p38 MAPK信号通路诱导AECOPD患者外周血中性粒细胞凋亡。
