李莘 李雪 王谙

基金项目：天津市教委科研计划项目（2022YGYB14）

作者单位：1天津市海河医院病理科（邮编300350）；2天津市呼吸疾病研究所；3天津市海河医院基础医学实验部

作者简介：李莘（1980），女，主任医师，主要从事呼吸系统疾病的诊断及治疗方面研究。E-mail：x_li2012@hotmail.com

摘要：目的 探讨温石棉对人脐静脉内皮细胞（HUVECs）的影响。方法 实验组以50、100、200 mg/L温石棉纤维液刺激HUVECs 24、48、72 h，对照组仅加RPMI 1640培养基培养细胞，观察细胞形态变化，β-半乳糖苷酶法分析细胞衰老情况，四甲基偶氮唑蓝法检测细胞存活率。采用实时荧光定量PCR法检测细胞中Wnt5a、p16和p21 mRNA的表达情况。结果 实验组HUVECs多呈梭形，部分呈圆形或不规则形，出现裸核及空泡现象，可见死亡细胞。随温石棉质量浓度及暴露时间的增加，细胞活性逐渐降低，衰老细胞逐渐增多。100 mg/L温石棉处理HUVECs 24 h时，细胞生长较活跃。与对照组相比，实验组Wnt5a、p16和p21 mRNA表达水平均增高（P＜0.05）。结论 温石棉可促进HUVECs衰老，Wnt5a、p16和p21参与此过程。

关键词：石棉，蛇纹石；人脐静脉内皮细胞；肺纤维化；Wnt-5a蛋白；基因，p16；基因，p21

中图分类号：R563.9文献标志码：ADOI：10.11958/20240060

Effects of chrysotile on expression of Wnt5a， p16 and p21 in endothelial cells

LI Xin1， 2， LI Xue2， 3， WANG An2， 3

1 Department of Pathology， Tianjin Haihe Hospital， Tianjin 300350， China; 2 Tianjin Key Laboratory of Lung Regenerative Medicine; 3 Tianjin Institute of Respiratory Diseases， Tianjin Haihe Hospital

Abstract： Objective To investigate the influence of chrysotile on human umbilical vein endothelial cells （HUVECs）. Methods HUVECs in the experimental group were treated with 50， 100 and 200 mg/L chrysotile for 24， 48 and 72 h. The control group was cultured with RPMI 1640 medium only. Morphological changes of HUVECs were observed. Cell viability was detected by MTT assay. β-Galactosidase staining was used to analyze the cell senescence. The cell survival rate was determined by tetramethylazolium blue assay. Wnt5a， p16 and p21 expression were investigated by qRT-PCR. Results Cells of the experimental group were oval and irregular， and some of them were round or irregular， with naked nuclei and vacuolar， and dead cells were present. With the increase of chrysotile concentration and exposure time， the level of cell viability decreased and the number of aged cells increased gradually （P＜0.05）. The cell growth of HUVECs was more active when 100 mg/L chrysotile treated for 24 h. Compared with the control group， the mRNA expression levels of Wnt5a， p16 and p21 were increased in the experimental group （P＜0.05）. Conclusion Chrysotile can promote HUVECs senescence， and Wnt5a， p16 and p21 are involved in this mechanism.

Key words： asbestos， serpentine; human umbilical vein endothelial cells; pulmonary fibrosis; Wnt-5a protein; genes， p16; genes， p21

我国为全球第一石棉消费国，目前90%以上使用的是温石棉［1-2］。长期大量暴露于石棉会导致石棉肺、间皮瘤、肺癌、喉癌等疾病的发生［3-4］。根据矿物学及化学结构，石棉纤维分为蛇纹石（温石棉）和角闪石两大类，温石棉在细胞内不稳定，易裂解被吞噬［5］，长期高浓度吸入会出现石棉肺等表现。石棉肺是指吸入过量石棉纤维导致的弥漫性肺纤维化［6］，目前针对肺纤维化的治疗方法有限。衰老受损肺组织的病理表现包括成纤维细胞增生、细胞外基质（ECM）沉积、血管结构破坏。……