中华皮肤科杂志 ›› 2025, Vol. 58 ›› Issue (2): 138-144.doi: 10.35541/cjd.20240475

• 论著 • 上一篇    下一篇

转化生长因子β1对婴幼儿血管瘤内皮细胞增殖、迁移及内皮间质转化的影响

龚雪    杨开颖    邱桐    向姗姗    周江元    吉毅   

  1. 四川大学华西医院小儿外科，成都  610041
    杨开颖现在广州医科大学附属妇女儿童医疗中心小儿外科，广州  510623

  • 收稿日期:2024-09-06 修回日期:2025-01-02 发布日期:2025-02-07
  • 通讯作者: 吉毅 E-mail:jijiyuanyuan@163.com
  • 基金资助:
    国家自然科学基金（82473553、82273556）；四川省杰出青年科学基金（2025NSFJQ0070）；四川大学从0到1创新研究项目（2022SCUH0033）；四川大学医学 + 信息中心交叉学科建设开放项目（YGJC004）；四川大学华西医院学科卓越发展1·3·5工程临床研究孵化项目（ZYJC21060）；四川大学华西医院临床研究基金（2023HXFH004）

Effect of transforming growth factor beta 1 on the proliferation, migration and endothelial-to-mesenchymal transition of infantile hemangioma-derived endothelial cells

Gong Xue, Yang Kaiying, Qiu Tong, Xiang Shanshan, Zhou Jiangyuan, Ji Yi   

  1. Department of Pediatric Surgery, West China Hospital, Sichuan University, Chengdu 610041, China
    Yang Kaiying is now working at the Department of Pediatric Surgery, Guangzhou Women and Children′s Medical Centre, Guangzhou Medical University, Guangzhou 510623, China
  • Received:2024-09-06 Revised:2025-01-02 Published:2025-02-07
  • Contact: Ji Yi E-mail:jijiyuanyuan@163.com
  • Supported by:
    National Natural Science Foundation of China （82473553, 82273556）; Excellent Youth Foundation of Sichuan Province of China （2025NSFJQ0070）; Project of ′0 to 1′ of Sichuan University （2022SCUH0033）; Med-X Center for Informatics Funding Project （YGJC004）; 1·3·5 Project for Disciplines of Excellence Project of West China Hospital of Sichuan University （ZYJC21060）; Clinical Research Fund of West China Hospital of Sichun University （2023HXFH004）

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摘要： 【摘要】 目的 研究转化生长因子β1（TGF-β1）对婴幼儿血管瘤（IH）内皮细胞（HemEC）生物活性的影响。方法 收集2021年2 - 8月在四川大学华西医院小儿外科行手术切除治疗的增殖期及消退期IH组织各3份，从增殖期IH组织中提取原代HemEC，以人脐静脉内皮细胞（HUVEC）为对照，通过免疫组化与Western印迹法分别检测TGF-β1在组织与细胞中的表达水平。采用细胞增殖毒性检测试剂盒（CCK8）检测0（对照组） ~ 100 ng/ml TGF-β1对HemEC细胞增殖的影响。采用5 ng/ml TGF-β1处理HemEC，以未接受TGF-β1处理为对照组，Transwell实验观察细胞迁移能力，免疫荧光检测内皮标志物血小板-内皮细胞黏附分子（CD31）、血管内皮钙黏蛋白（VE-cadherin）及间质标志物α平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原α1（COL1A1）的表达变化。组间比较采用两独立样本t检验或单因素方差分析。结果 免疫组化结果显示，增殖期IH组织TGF-β1染色阳性且表达相对丰富，阳性信号面积占比为24.68% ± 3.74%，而消退期IH组织中几乎不表达TGF-β1。Western印迹实验显示，HemEC中TGF-β1相对表达水平（1.08 ± 0.13）高于HUVEC（0.30 ± 0.04，t = 9.93，P < 0.001）。CCK8实验显示，3.125、6.25、12.5、25、50、75 ng/ml TGF-β1组HemEC增殖活性均高于对照组（均P < 0.05），而100 ng/ml TGF-β1组与对照组细胞增殖活性差异无统计学意义（P > 0.05）。Transwell实验显示，5 ng/ml TGF-β1组细胞迁移数［（127 ± 6）个］高于对照组［（103 ± 9）个］，t = 5.32，P < 0.01。HemEC免疫荧光检测显示，5 ng/ml TGF-β1组CD31、VE-cadherin荧光强度（5.441 ± 1.254、5.073 ± 0.412）均低于对照组（9.518 ± 1.728、7.671 ± 0.921），差异有统计学意义（t = 3.31、4.46，P = 0.030、0.011）；α-SMA、COL1A1荧光强度（8.074 ± 0.846、5.885 ± 0.216）高于对照组（0.393 ± 0.342、0.295 ± 0.125），差异有统计学意义（t = 14.58、38.76，P < 0.001、 < 0.000 1）。结论 TGF-β1在增殖期IH组织与HemEC中高表达，并可促进HemEC增殖、迁移及内皮间质转化。

关键词: 血管瘤, 转化生长因子β1, 细胞增殖, 细胞迁移分析, 血管瘤内皮细胞, 内皮间质转化

Abstract: 【Abstract】 Objective To investigate the effect of transforming growth factor beta 1 （TGF-β1） on the biological activity of infantile hemangioma （IH）-derived endothelial cells （HemECs）. Methods Three proliferating IH tissues and three involuting IH tissues were collected from IH patients receiving surgical resection at the Department of Pediatric Surgery, West China Hospital, Sichuan University from February to August 2021. Primary HemECs were isolated from proliferating IH tissues, and human umbilical vein endothelial cells （HUVECs） served as the control. The TGF-β1 expression levels in tissues and cells were detected by immunohistochemical study and Western blot analysis. Cell counting kit-8（CCK8）assay was performed to assess the effect of 0 （control group） - 100 ng/ml TGF-β1 on HemEC proliferation. HemECs were treated with 5 ng/ml TGF-β1 or without （control group）, and after several hours of treatment, Transwell assay was performed to evaluate cell migration ability, and immunofluorescence assay to assess the changes in the expression of endothelial markers （platelet-endothelial cell adhesion molecule-1 ［CD31］, vascular endothelial cadherin ［VE-cadherin］） and mesenchymal markers （α-smooth muscle actin ［α-SMA］, collagen type Ⅰ α 1 ［COL1A1］）. Comparisons between groups were conducted by t test or one-way analysis of variance. Results Immunohistochemical study showed that proliferating IH tissues were stained positively for TGF-β1, which was expressed relatively abundantly; the percentages of TGF-β1-positive signal area were higher in the proliferating IH tissues （24.68% ± 3.74%） than in the involuting IH tissues （almost no expression）. Western blot analysis revealed that the relative expression level of TGF-β1 was significantly higher in HemECs （1.08 ± 0.13） than in HUVECs （0.30 ± 0.04, t = 9.93, P < 0.001）. CCK8 assay showed increased proliferative activity of HemECs in the 3.125-, 6.25-, 12.5-, 25-, 50- and 75-ng/ml TGF-β1 groups compared with the control group （all P < 0.05）, and no significant difference was found between the 100-ng/ml TGF-β1 group and the control group （P > 0.05）. Transwell assay revealed an increased number of migratory HemECs in the 5-ng/ml TGF-β1 group （127 ± 6） compared with the control group （103 ± 9; t = 5.32, P < 0.01）. Immunofluorescence assay showed significantly decreased fluorescence intensity of endothelial markers CD31 and VE-cadherin in the 5-ng/ml TGF-β1 group （5.441 ± 1.254, 5.073 ± 0.412, respectively） compared with the control group （9.518 ± 1.728,7.671 ± 0.921, t = 3.31, 4.46, P = 0.030, 0.011, respectively）, and significantly increased fluorescence intensity of mesenchymal markers α-SMA and COL1A1 in the 5-ng/ml TGF-β1 group （8.074 ± 0.846, 5.885 ± 0.216, respectively） compared with the control group （0.393 ± 0.342, 0.295 ± 0.125, t = 14.58, 38.76, P < 0.001, < 0.000 1, respectively）. Conclusion TGF-β1 was relatively highly expressed in the proliferating IH tissues and HemECs, and could promote the proliferation, migration and endothelial-to-mesenchymal transition of HemECs.

Key words: Hemangioma, Transforming growth factor beta 1, Cell proliferation, Cell migration assays, Hemangioma-derived endothelial cells, Endothelial-to-mesenchymal transition

引用本文

龚雪 杨开颖 邱桐 向姗姗 周江元 吉毅. 转化生长因子β1对婴幼儿血管瘤内皮细胞增殖、迁移及内皮间质转化的影响[J]. 中华皮肤科杂志, 2025,58(2):138-144. doi:10.35541/cjd.20240475

Gong Xue, Yang Kaiying, Qiu Tong, Xiang Shanshan, Zhou Jiangyuan, Ji Yi. Effect of transforming growth factor beta 1 on the proliferation, migration and endothelial-to-mesenchymal transition of infantile hemangioma-derived endothelial cells[J]. Chinese Journal of Dermatology, 2025, 58(2): 138-144.doi:10.35541/cjd.20240475