Xu Hui, Yang Jingwen, Tu Mengyun, Weng Jie, Xie Mengying, Zhou Zhiliang, Zhou Peisen, Wang Liang, Chen Chan, Wang Zhiyi
Department of General Practice, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
Department of Geriatric Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
Front Pharmacol. 2022 May 9;13:901000. doi: 10.3389/fphar.2022.901000. eCollection 2022.
Vincristine (VCR) is used in the clinic as an anti-tumor drug. VCR can cause pulmonary fibrosis (PF), leading to respiratory failure. The transformation of fibroblasts into myofibroblasts may play a key role in PF. The present study attempted to reveal the molecular mechanism of VCR-induced PF and the possible involvement of the mitogen-activated protein kinase (MAPK) signaling pathway. Human embryonic lung fibroblasts (HELFs) were treated with different concentrations of VCR. Inhibitors of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 MAPK were added to HELFs. Cell proliferation state was assessed using cell counting kit-8 and by directly counting the number of cells. The expressions of vimentin and -smooth muscle actin (α-SMA) were investigated using western blot and immunofluorescence analyses. Activation of ERK and P38 was estimated by the expression of phosphorylated p38 MAPK (p-p38), p38 MAPK, phosphorylated ERK1/2 (p-ERK1/2) and ERK1/2 using western blot analysis. Enzyme-linked immunosorbent assay was used to estimate the level of collagen I in cell culture supernatants. Results showed that VCR promoted cellular proliferation, secretion of collagen I and the expression of vimentin and -SMA. High expression of p-p38 and p-ERK1/2 was associated with the activation of the MAPK signaling pathway. MAPK inhibitors SB203580 and PD98059 suppressed the expression of the above proteins. Our study revealed that VCR could promote the differentiation of fibroblasts into myofibroblasts by regulating the MAPK signal pathway, which may be a promising way to treat VCR-induced PF.
长春新碱（VCR）在临床上用作抗肿瘤药物。VCR可导致肺纤维化（PF），进而引发呼吸衰竭。成纤维细胞向肌成纤维细胞的转化可能在PF中起关键作用。本研究试图揭示VCR诱导PF的分子机制以及丝裂原活化蛋白激酶（MAPK）信号通路可能的参与情况。用人胚胎肺成纤维细胞（HELFs）进行不同浓度VCR处理。向HELFs中加入细胞外信号调节激酶1/2（ERK1/2）和p38 MAPK的抑制剂。使用细胞计数试剂盒-8并直接计数细胞数量来评估细胞增殖状态。采用蛋白质印迹法和免疫荧光分析法研究波形蛋白和α平滑肌肌动蛋白（α-SMA）的表达。通过蛋白质印迹分析磷酸化p38 MAPK（p-p38）、p38 MAPK、磷酸化ERK1/2（p-ERK1/2）和ERK1/2的表达来评估ERK和P38的激活情况。采用酶联免疫吸附测定法评估细胞培养上清液中I型胶原蛋白的水平。结果显示，VCR促进细胞增殖、I型胶原蛋白分泌以及波形蛋白和α-SMA的表达。p-p38和p-ERK1/2的高表达与MAPK信号通路的激活相关。MAPK抑制剂SB203580和PD98059抑制上述蛋白的表达。我们的研究表明，VCR可通过调节MAPK信号通路促进成纤维细胞向肌成纤维细胞分化，这可能是治疗VCR诱导的PF的一种有前景的方法。
