低温等离子体活化液对血管内皮细胞增殖和血管生成能力的影响  

Effects of low temperature plasma-activated medium on proliferation and angiogenic capacity of vascular endothelial cells

作　　者：袁网[1] 王香妮 刘进仁 王曦迎 鹿佳佳 何芷柔 许玉琳 石兴民[2] YUAN Wang;WANG Xiangni;LIU Jinren;WANG Xiying;LU Jiajia;HE Zhirou;XU Yulin;SHI Xingmin(PET-CT Room,The First Affiliated Hospital of Xi'an Jiaotong University,Xi’an 710061;School of Public Health,Xi'an Jiaotong University,Xi’an 710061;Yuncheng Vocational and Technical University,Yuncheng 044000,China)

机构地区：[1]西安交通大学第一附属医院PET-CT室,陕西西安710061 [2]西安交通大学公共卫生学院,陕西西安710061 [3]运城职业技术大学,山西运城044000

出　　处：《西安交通大学学报（医学版）》2024年第3期388-395,共8页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省自然科学基础研究计划面上项目(No.2022JM-556);陕西省青年基金项目(No.2022JQ-882);陕西省中药管理局科研项目(No.SZY-KJCYC-2023-084)。

摘　　要：目的探索低温等离子体(low temperature plasma,LTP)产生的活化液(plasma-activated medium,PAM)对人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖和血管生成的影响,为未来利用PAM促进创伤愈合和抑制肿瘤血管生成提供理论基础。方法选择HUVECs作为体外研究模型,以LTP处理不同时间(0、15、30、45、60、75 s)后的PAM培养液进行分组干预。采用MTT法和细胞周期法测定PAM对HUVECs活力的影响,通过血管生成实验检测PAM对血管生成的影响,采用荧光探针检测细胞内活性氧(ROS)水平;构建黑色素瘤小鼠模型,免疫组织化学法检测肿瘤组织中CD31的表达以评估血管生成情况。结果LTP处理获得的PAM对HUVECs血管生成具有双向作用,表现为:与对照组(0 s)相比,低剂量(15 s和30 s)处理时,HUVEC细胞活力增加,而高剂量(45 s、60 s和75 s)处理时,细胞活力明显下降(P<0.05);PAM-15 s和PAM-30 s组处于S期的细胞比例明显增高,而PAM-45 s、PAM-60 s、PAM-75 s组处于S期细胞比例显著降低,差异具有统计学意义(P<0.05);PAM-15 s和PAM-30 s组HUVEC血管生成能力增强,而PAM-45 s、PAM-60 s、PAM-75 s组的细胞血管生成能力降低,血管节点、交叉点、网眼数和血管分支数都明显降低(P<0.05)。此外,HUVECs内ROS水平随着LTP处理时间的延长而升高。PAM处理黑色素瘤小鼠后,对照组肿瘤组织中CD31棕黄色阳性信号大面积分布,各PAM处理组肿瘤组织中CD31棕黄色颗粒分布减少。结论短时间LTP处理的PAM促进了HUVEC的增殖和血管生成;而长时间LTP处理的PAM则抑制细胞活力和血管生成。Objective To explore the plasma-activated medium(PAM)produced by low temperature plasma(LTP)on the proliferation and angiogenesis of human umbilical vein endothelial cells(HUVECs)so as to provide theoretical basis for the future use of PAM to promote wound healing and inhibit tumor angiogenesis.Methods HUVECs were selected as the in vitro research model.The PAM-containing medium after LTP treatment for different time points(0 s,15 s,30 s,45 s,60 s,and 75 s)was used for intervention.The influence of PAM on HUVECs viability was assessed using the MTT assay and cell cycle analysis.The effects of PAM on angiogenesis were examined through angiogenesis experiments.Intracellular levels of reactive oxygen species(ROS)were measured using fluorescence probes.A melanoma mouse model was established,and CD31 expression was detected by immunohistochemistry.Results As the treatment time increased,the intracellular levels of ROS also elevated.PAM derived from LTP exhibited a bidirectional effect on angiogenesis in HUVECs.Compared to the control group(0 s),low-dose treatments(15 s and 30 s)enhanced HUVECs viability,while high-dose treatments(45 s,60 s,and 75 s)significantly decreased cell viability(P<0.05).The proportion of HUVECs in the S phase was significantly increased in the PAM-15 s and PAM-30 s groups,but markedly decreased in the PAM-45 s,PAM-60 s,and PAM-75 s groups,with statistically significant differences(P<0.05).The HUVECs tube formation ability was enhanced in the 15 s and 30 s PAM groups,but diminished in the PAM-45 s,PAM-60 s,and PAM-75 s groups,characterized by the decreased numbers of vascular nodes,intersections,meshes,and branching points(P<0.05).After PAM treatment in the melanoma mouse model,the control group exhibited widespread distribution of CD31 in tumor tissue,while the PAM-5 min and PAM-10 min groups displayed reduced distribution of CD31.Conclusion Short-term exposure to PAM enhances HUVECs proliferation and angiogenesis,whereas prolonged exposure suppresses cell viability and inhibits angiogenesis.

关 键 词：低温等离子体 血管内皮细胞 血管生成 

分 类 号：R333.12[医药卫生—人体生理学]