摘要
目的:探讨姜黄素对婴儿血管瘤内皮细胞(HemECs)增殖、凋亡的影响及机制。方法:分离培养HemECs,给予不同浓度姜黄素,CCK-8法检测HemECs增殖,台盼蓝法检测HemECs死亡率;流式细胞仪检测细胞凋亡;RT-qPCR法检测HIF-1αmRNA表达,电镜观察细胞凋亡;免疫荧光检测HIF-1α和VEGF表达,Western blot检测MCL-1、cl-Caspase-3和cl-PARP蛋白表达。结果:48 h后不同浓度姜黄素均可抑制HemECs增殖,且呈剂量依赖性。对照组和姜黄素组HemECs凋亡率差异有统计学意义(P<0.05);姜黄素处理后HemECs超微结构出现核碎裂、染色质浓缩以及核膜消失等特征;姜黄素处理48 h后,HIF-1α和VEGF蛋白表达下调,HIF-1αmRNA表达下调(P<0.05);姜黄素组MCL-1蛋白表达水平明显低于对照组,而cl-Caspase-3和cl-PARP蛋白表达升高。结论:姜黄素可抑制HemECs增殖,诱导其凋亡,且呈剂量依赖性,其机制可能是下调MCL-1和HIF-1α表达,上调cl-Caspase-3和cl-PARP表达。
Objective:To investigate effect of curcumin on proliferation and apoptosis of Hemangioma endothelial cells(HemECs)in infants and its mechanism.Methods:HemECs were isolated and cultured and treated with different concentrations of curcumin.Proliferation of HemECs was detected by CCK-8 method,and mortality of HemECs was detected by trypan blue method.Apoptotic cells were detected by flow cytometry,relative expression of HIF-1αmRNA was detected by RT-qPCR,and apoptotic cells were observed by electron microscopy.Expressions of HIF-1αand VEGF were detected by immunofluorescence and expressions of MCL-1,cl-Caspase-3 and cl-PARP were detected by Western blot.Results:Curcumin at different concentrations inhibited HemECs proliferation 48 h later in a dose-dependent manner.Apoptotic rate of HemECs in control group and curcumin group had significant difference(P<0.05).After curcumin treatment,ultrastructure of HemECs showed nuclear fragmentation,chromatin concentration and nuclear membrane disappearance.Curcumin could down-regulate expressions of HIF-1αand EGF after 48 h,inhibit expression of HIF-1αmRNA(P<0.05).Expressions of MCL-1 protein in curcumin group was significantly lower than that in control group,while expressions of cl-Caspase 3 and cl-PARP proteins were increased.Conclusion:Curcumin can inhibit proliferation and induce apoptosis of HemECs in a dose-dependent manner,whose mechanism may be down-regulation of MCL-1 and HIF-1αexpressions and up-regulation of cl-Caspase-3 and cl-PARP expressions.
作者
符垂师
云川
冯业成
黄天
FU Chui-Shi;YUN Chuan;FENG Ye-Cheng;HUANG Tian(Department of Pediatrics,First Affiliated Hospital of Hainan Medical College,Haikou 570102,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第17期2086-2090,共5页
Chinese Journal of Immunology
基金
海南省卫生和计划生育委员会计划项目(No.1601033032A2001)资助。
