天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (12): 1240-1244.doi: 10.11958/20211483

• 细胞与分子生物学 • 上一篇    下一篇

低氧对HTR-8/SVneo细胞增殖及HIF-1α、VEGF、MMP-9、TIMP-1表达的影响

袁烁 1，刘湘云 2，张家旗 3，邓高丕 1△   

  1. 1广州中医药大学第一附属医院妇科（邮编510405）；2佛山市中医院；3广州中医药大学
  • 收稿日期:2021-06-22 修回日期:2021-08-02 出版日期:2021-12-15 发布日期:2021-12-27
  • 通讯作者: 通信作者 E-mail：denggaopi@126.com E-mail:denggaopi@126.com
  • 作者简介:袁烁（1982），女，博士，副主任中医师，主要从事生殖障碍的中西医结合治疗研究。E-mail：twinklingyuan@126.com
  • 基金资助:
    国家自然科学基金资助项目（81804134）；广东省自然科学基金资助项目（2018A030313469）；2019年广东省名中医传承工作室 建设项目（邓高丕广东省名中医传承工作室）［粤中医办函〔2019〕5号］

Effects of hypoxia on proliferation and the expression of HIF-1α, VEGF, MMP-9 and TIMP-1 in HTR-8/SVneo cells

YUAN Shuo1, LIU Xiang-yun2, ZHANG Jia-qi3, DENG Gao-pi1△   

  1. 1 Department of Gynecology, the First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, China; 2 Foshan Hospital of Chinese Medicine; 3 Guangzhou University of Chinese Medicine △Corresponding Author E-mail: denggaopi@126.com
  • Received:2021-06-22 Revised:2021-08-02 Published:2021-12-15 Online:2021-12-27

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摘要： 目的 采用二氯化钴（CoCl2）模拟体外低氧环境，探讨低氧状态对滋养细胞株HTR-8/SVneo细胞增殖及缺 氧诱导因子（HIF）-1α、血管内皮生长因子（VEGF）、基质金属蛋白酶（MMP）-9、基质金属蛋白酶组织抑制因子 1 （TIMP-1）蛋白表达的影响。方法 采用0、50、100、200、400、800、1 000、1 200 μmol/L的CoCl2处理HTR-8/SVneo细 胞，建立化学缺氧模型，CCK8法检测以上浓度培养24 h、48 h后对HTR-8/SVneo细胞增殖的影响；根据CCK8实验结 果拟定 CoCl2低、中、高浓度组，Western blot 检测 CoCl2低、中、高浓度组作用 48 h 对 HTR-8/SVneo 细胞中 HIF-1α、 VEGF、MMP-9、TIMP-1蛋白表达的影响。结果 CCK8实验提示低氧状态激活HTR-8/SVneo细胞增殖，CoCl2作用于 HTR-8/SVneo细胞48 h后，OD值随浓度增加而上升；同一CoCl2浓度作用下，作用48 h较作用24 h的OD值均上升。 选择100、200、400 μmol/L的CoCl2作为CoCl2低、中、高浓度组。与空白组对比，CoCl2低、中、高浓度组HTR-8/SVneo 细胞中HIF-1α、MMP-9蛋白表达水平上调（P＜0.05），MMP-9/TIMP-1比例上升，呈浓度依赖性（P＜0.05），CoCl2中、 高浓度组的VEGF的蛋白表达水平上调（P＜0.05），CoCl2低、中、高浓度组的TIMP-1蛋白表达水平下降，但随CoCl2浓 度增加而呈上升趋势。结论 低氧状态可增强 HTR-8/SVneo 细胞的增殖能力，并可能通过 HIF-1α 的介导上调 VEGF的表达与MMP-9/TIMP-1比值。

关键词: 低氧, 细胞增殖, 缺氧诱导因子1, α亚基, 基质金属蛋白酶9, 血管内皮生长因子类, 滋养细胞

Abstract: Objective To investigate the effects of hypoxia on the proliferation of trophoblast cell line HTR-8/SVneo cells and the protein expression of hypoxia inducer (HIF) -1α, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP) -9 and tissue inhibitor of matrix metalloproteinase 1 (TIMP-1). Methods HTR-8/SVneo cells were treated with 0, 50, 100, 200, 400, 800, 1 000, 1 200 μmol/L CoCl2 to establish the chemical hypoxia model. The proliferation of HTR-8/SVneo cells was detected by CCK8 method after 24 h and 48 h culture at the above concentration. According to the results of CCK8 experiment, the hypoxic, medium and high concentration groups were determined. Western blot assay was used to detect changes of the protein expression levels of HIF-1α, VEGF, MMP-9 and TIMP-1 in HTR-8/ SVneo cells in the hypoxic, medium and high concentration groups. Results CCK8 experiment showed that hypoxia activated the proliferation of HTR-8/SVneo cells. After treatment with CoCl2 on HTR-8/SVneo cells for 48 hours, the OD value increased with the increase of concentration. Under the same CoCl 2 concentration, the OD value of 48 h was higher than that of 24 h. The 100, 200 and 400 μmol/ L CoCl2 were used as the low, medium and high concentration groups. After 48 hours, compared with the blank group, the expression levels of HIF-1α, MMP-9 protein in HTR-8/SVneo cells were upregulated in the low, medium and high concentration groups (P＜0.05). The ratio of MMP-9/TIMP-1 increased in a concentration dependent manner (P＜0.05). The protein expression of VEGF was upregulated in the hypoxia, medium and high concentration groups (P＜0.05). The expression level of TIMP-1 protein decreased in the low, medium and high CoCl2 groups, but it showed an increased trend with the increased CoCl2 concentrations. Conclusion Hypoxia can enhance the proliferation of HTR-8/SVneo cells, and the expression of VEGF and the ratio of MMP-9/TIMP-1 can be upregulated by HIF-1α.

Key words: hypoxia, cell proliferation, hypoxia-inducible factor 1, alpha subunit, matrix metalloproteinase 9, vascular endothelial growth factors, trophocyte

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