摘要
目的采用二氯化钴(CoCl_(2))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 200μmol/L的CoCl_(2)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CoCl_(2)低、中、高浓度组,Western blot检测CoCl_(2)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,CoCl_(2)作用于HTR-8/SVneo细胞48 h后,OD值随浓度增加而上升;同一CoCl_(2)浓度作用下,作用48 h较作用24 h的OD值均上升。选择100、200、400μmol/L的CoCl_(2)作为CoCl_(2)低、中、高浓度组。与空白组对比,CoCl_(2)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),CoCl_(2)中、高浓度组的VEGF的蛋白表达水平上调(P<0.05),CoCl_(2)低、中、高浓度组的TIMP-1蛋白表达水平下降,但随CoCl_(2)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。
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 CoCl_(2) 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 CoCl_(2) 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 CoCl_(2) were used as the low,medium and high concentration groups.After48 hours,compared with the blank group,the expression levels of HIF-1α,MMP-9 protein in HTR-8/SVneo cells were up regulated 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 CoCl_(2) groups,but it showed an increased trend with the increased CoCl_(2) 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.
作者
袁烁
刘湘云
张家旗
邓高丕
YUAN Shuo;LIU Xiang-yun;ZHANG Jia-qi;DENG Gao-pi(Department of Gynecology,the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Foshan Hospital of Chinese Medicine;Guangzhou University of Chinese Medicine)
出处
《天津医药》
CAS
北大核心
2021年第12期1240-1244,共5页
Tianjin Medical Journal
基金
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]。
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
hypoxia
cell proliferation
hypoxia-inducible factor 1,alpha subunit
matrix metalloproteinase 9
vascular endothelial growth factors
trophocyte
