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中华普通外科学文献(电子版) ›› 2025, Vol. 19 ›› Issue (03) : 169 -174. doi: 10.3877/cma.j.issn.1674-0793.2025.03.004

论著

缺氧对人大隐静脉平滑肌细胞行为特征的影响
居建华1, 周顺昌1, 徐永波2, 胡晓璇3, 钟玉绪3, 褚海波1,()   
  1. 1. 266300 青岛,同济大学附属东方医院胶州医院普通外科
    2. 261021 潍坊,中国人民解放军陆军第80集团军医院普通外科
    3. 100850 北京,中国人民解放军军事科学院军事医学研究院毒物药物研究所
  • 收稿日期:2024-11-06 出版日期:2025-06-01
  • 通信作者: 褚海波
  • 基金资助:
    中国人民解放军后勤重大项目 (AWS17J008)

Effect of behavioral characteristics in smooth muscle cells deriving from the human great saphenous veins underlying hypoxia

Jianhua Ju1, Shunchang Zhou1, Yongbo Xu2, Xiaoxuan Hu3, Yuxu Zhong3, Haibo Chu1,()   

  1. 1. Department of General Surgery, Jiaozhou Branch of Shanghai East Hospital, Tongji University, Qingdao 266300, China
    2. Department of General Surgery, the 80th Group Army Hospital of People’s Liberation Army, Weifang 261021, China
    3. State Key Laboratory of Toxicology and Medical Countermeasures, Institute of Toxicology and Pharmacology , Beijing 100850, China
  • Received:2024-11-06 Published:2025-06-01
  • Corresponding author: Haibo Chu
引用本文:

居建华, 周顺昌, 徐永波, 胡晓璇, 钟玉绪, 褚海波. 缺氧对人大隐静脉平滑肌细胞行为特征的影响[J/OL]. 中华普通外科学文献(电子版), 2025, 19(03): 169-174.

Jianhua Ju, Shunchang Zhou, Yongbo Xu, Xiaoxuan Hu, Yuxu Zhong, Haibo Chu. Effect of behavioral characteristics in smooth muscle cells deriving from the human great saphenous veins underlying hypoxia[J/OL]. Chinese Archives of General Surgery(Electronic Edition), 2025, 19(03): 169-174.

目的

通过建立静脉平滑肌细胞(SMCs)模拟缺氧模型,探讨缺氧环境下人大隐静脉(GSVs)管壁SMCs 的行为特征。

方法

收集2020 年5 月至2021 年5 月中国人民解放军陆军第80 集团军医院收治的15 例冠状动脉搭桥而GSVs 无损伤患者的正常GSVs 组织样本,分离和培养SMCs 后,分为二氯化钴(CoCl2)组和对照组。采用多种染色法观察SMCs 增殖、迁移、黏附、衰老及细胞骨架结构;聚合酶链反应和免疫印迹法检测Bax、Bcl-2、caspase-3、MMP-2、MMP-9、TIMP-1、TIMP-2 mRNA 和蛋白表达。

结果

CoCl2 组SMCs 黏附、迁移、衰老、增殖数量比对照组明显增多(t=5.994、5.687、13.898、25.979;均P<0.05);CoCl2 组Bax、caspase-3 mRNA 和蛋白表达比对照组明显降低(t=27.297、15.430、17.851、17.801;均P<0.05);而Bcl-2、MMP-2、MMP-9、TIMP-1、TIMP-2 mRNA 和蛋白表达较对照组明显升高(t=11.704、20.705、16.363、12.228、11.473、9.618、14.780、14.691、64.185、14.915;均P<0.05)。

结论

在缺氧条件下,人GSVs 管壁中SMCs 的表型与功能异常,提示SMCs 从收缩型表型向分泌型或合成型表型转变,呈去分化状态;经线粒体通路细胞凋亡数减少。

Objective

To establish a simulated hypoxia model of venous smooth muscle cells(SMCs), and to investigate the effect of hypoxia on the behaviorial characteristics of SMCs derived from the human normal great saphenous veins (GSVs).

Methods

Fifteen tissue samples were collected from patients undergoing distal arterial bypass grafting surgery without GSVs injuries in the 80th Group Army Hospital of the People’s Liberation Army from May 2020 to May 2021.SMCs were isolated and cultured, then the cobalt chloride (CoCl2) group and the control group were assigned.Using specific staining, proliferation,migration, adhesion, senescence and the structure of cytoskeletal filaments in SMCs were observed.mRNA and protein expression of Bax, Bcl-2, caspase-3, matrix metalloproteinases (MMP)-2, MMP-9, tissue inhibitor of metalloproteinases (TIMP)-1, and TIMP-2 were detected by fluorescent quantitative polymerase chain reaction and immunoblotting.

Results

The quantity of adhesion, migration, senescence, proliferation and mRNA and protein expression of Bcl-2, MMP-2, MMP-9, TIMP-1, and TIMP-2 in SMCs in the CoCl2 group were significantly increased compared with the control group (t=5.994, 5.687, 13.898, 25.979,11.704, 20.705, 16.363, 12.228, 11.473, 9.618, 14.780, 14.691, 64.185, 14.915, all P<0.05).mRNA and protein expression of Bax and caspase-3 were significantly decreased (t=27.297, 15.430, 17.851, 17.801, all P<0.05).

Conclusion

Under hypoxic conditions, the phenotype and function of SMCs derived from the human NGSVs are dysregulated, suggesting that SMCs switch from the contractile phenotype to the secretory or synthetic phenotype, and more dedifferentiate, and apoptotic decrease through the intrinsic pathway.

表1 相关基因引物序列及扩增产物大小
图1 正常血管平滑肌细胞(VSMCs)表型A:VSMCs形成细胞晕(×40);B:VSMCs 呈长梭形或多角形(×100);C:VSMCs 呈“峰谷”状生长(×40);D:α-SMA 免疫荧光染色阳性(×400)
图2 两组黏附的平滑肌细胞分布(×100)与数量*P<0.001
图3 两组迁移的平滑肌细胞分布(DAPI 染色 ×100)与数量*P<0.001
图4 两组衰老的平滑肌细胞分布(β-gal 染色×100)与数量*P<0.001
图5 两组增殖的平滑肌细胞数量变化 *P<0.001
图6 两组平滑肌细胞骨架(F-actin 荧光染色×400)
图7 两组血管平滑肌细胞 Bax、Bcl-2、caspase-3 mRNA和蛋白表达 A: Bax mRNA 表达;B: Bcl-2 mRNA 表达;C: caspase-3 mRNA 表达;D:Bax、Bcl-2、caspase-3、β-actin 蛋白条带;E: Bax 蛋白表达;F: Bcl-2 蛋白表达;G: caspase-3 蛋白表达;与对照组比较,*P<0.05
表2 两组平滑肌细胞Bax、Bcl-2、caspase-3 mRNA 和蛋白表达比较(n=15,)
图8 两组血管平滑肌细胞MMP-2、MMP-9、TIMP-1、TIMP-2 mRNA 和蛋白表达 A: MMP-2 mRNA 表达;B:MMP-9 mRNA 表达;C: TIMP-1 mRNA 表达;D: TIMP-2 mRNA 表达;E: MMP-2、MMP-9、TIMP-1、TIMP-2、βactin 蛋白条带;F: MMP-2 蛋白表达;G: MMP-9 蛋白表达;H: TIMP-1 蛋白表达;I: TIMP-2 蛋白表达;与对照组比较,*P<0.05
表3 两组平滑肌细胞MMP-2、MMP-9、TIMP-1、TIMP-2 mRNA 和蛋白表达比较(n=15, )
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