VEGF在肺纤维化肺组织内的表达及其作用研究
Study on the expression and effect of VEGF in the pulmonary fibrosis
-
摘要: 目的：探讨血管内皮生长因子（VEGF）在肺纤维化病人及模型小鼠内的表达及其在肺纤维化疾病中发挥的作用。方法：收集临床确诊为肺纤维化的病人及正常供体肺组织，通过免疫组织化学染色检测VEGF和α-平滑肌肌动蛋白（α-SMA）的表达，统计VEGF阳性表达与肺纤维化严重程度的相关性。给予小鼠气管注射博来霉素21 d诱导肺纤维化模型，免疫组织化学染色检测模型小鼠肺组织内VEGF和α-SMA的表达水平。给予小鼠气管注射VEGF164抗体后，给予博来霉素诱导小鼠肺纤维化模型，H&E染色和Masson三色染色比较小鼠肺组织纤维化病理损伤和胶原表达，并进行Ashcroft评分，使用荧光定量PCR比较肺组织内α-SMA和纤连蛋白的表达水平变化，使用试剂盒检测肺组织内羟脯氨酸含量的变化。结果：VEGF在肺纤维化病人和小鼠肺组织内明显的阳性表达，且阳性表达率越高，α-SMA表达水平越高，呈显著的相关性。给予VEGF164抗体阻断小鼠肺组织内VEGF表达后，小鼠肺纤维化病理损伤明显减轻，胶原含量减少，α-SMA和纤连蛋白的mRNA水平降低，羟脯氨酸含量减少。结论：VEGF在肺纤维化病人和小鼠肺组织内呈显著的阳性表达，阻断肺组织内VEGF表达缓解了博来霉素诱导的肺纤维化。Abstract: Objective:To determine the expression of VEGF in pulmonary fibrosis patients and mice model,and its effect on disease.Methods:The expression levels of VEGF and α-SMA in lung tissue of pulmonary fibrosis patients and normal lung tissue were detected using immunohistochemical staining,and the correlation of positive expression of VEGF with severity of pulmonary fibrosis was analyzed.The mice model with pulmonary fibrosis was established using bleomycin injecting trachea of mice for 21 d,and the expression levels of VEGF and α-SMA in lung tissue of mice were detected using immunohistochemical staining.After treatment with VEGF164 antibody,the pathological damage and collagen expression in lung tissue of pulmonary fibrosis mice induced by bleomycin were analyzed using H&E and Masson staining,the lung tissue was evaluated using Ashcroft score,the mRNA levels of α-SMA and fibronectin in lung tissue were measured by Real-time PCR,and the changes of hydroxyproline content in lung tissue was detected using kit.Results:The positive expression of VEGF in lung tissue of patients with pulmonary fibrosis and bleomycin-induced mice were identified,and the expression of α-SMA was correlative with VEGF positive rate.After VEGF164 antibody blocking the expression of VEGF in lung tissue,the pathological damage and collagen expression in lung tissue of pulmonary fibrosis significantly alleviated and decreased,the mRNA levels of α-SMA and fibronectin decreased,and the changes of hydroxyproline content decreased.Conclusions:The expression of VEGF in patients with pulmonary fibrosis and bleomycin-induced pulmonary fibrosis mice are positive,and blocking the VEGF expression can relieve the PF induced by BLM.
-
Key words:
- pulmonary fibrosis /
- VEGF /
- α-SMA
-
[1] O'DWYER DN,ASHLEY SL,MOORE BB.Influences of innate immunity,autophagy,and fibroblast activation in the pathogenesis of lung fibrosis[J].Am J Physiol Lung Cell Mol Physiol,2016,311(3):L590. [2] KOLB M,BONELLA F,WOLLIN L.Therapeutic targets in idiopathic pulmonary fibrosis[J].Respir Med,2017(131):49. [3] HUANG H,PENG X,ZHONG C.Idiopathic pulmonary fibrosis:The current status of its epidemiology,diagnosis,and treatment in China[J].Intractable Rare Dis Res,2013,2(3):88. [4] LOOMIS-KING H,FLAHERTY KR,MOORE BB.Pathogenesis,current treatments and future directions for idiopathic pulmonary fibrosis[J].Curr Opin Pharmacol,2013,13(3):377. [5] BARRATT SL,FLOWER VA,PAULING JD,et al.VEGF (vascular endothelial growth factor) and fibrotic lung disease[J].Int J Mol Sci,2018,19(5).pii:E1269. [6] SPAGNOLO P,TZOUVELEKIS A,BONELLA F.The management of patients with idiopathic pulmonary fibrosis[J].Front Med,2018(5):148. [7] TAKEDA T,TAKEUCHI M,SAITOH M,et al.Improvement in patient-reported outcomes and forced vital capacity during nintedanib treatment of idiopathic pulmonary fibrosis[J].Tohoku J Exp Med,2018,245(2):107. [8] TZOUVELEKIS A,ANEVLAVIS S,BOUROS D.Angiogenesis in interstitial lung diseases:a pathogenetic hallmark or a bystander?[J].Respir Res,2006(7):82. [9] BARRATT SL,BLYTHE T,JARRETT C,et al.Differential expression of VEGF-Axxx isoforms is critical for development of pulmonary fibrosis[J].Am J Respir Crit Care Med,2017,196(4):479. [10] MOELLER A,ASK K,WARBURTON D,et al.The bleomycin animal model:a useful tool to investigate treatment options for idiopathic pulmonary fibrosis[J].Int J Biochem Cell Biol,2008,40(3):362. -
点击查看大图
计量
- 文章访问数: 3778
- HTML全文浏览量: 516
- PDF下载量: 48
- 被引次数: 0
下载:
