小檗碱对人脐静脉内皮细胞增殖与凋亡的作用

Effects of berberine on proliferation and apoptosis of HUVECs

目的:观察小檗碱对人脐静脉内皮细胞增殖与凋亡的作用,以探讨小檗碱抑制肿瘤生长与转移的机制。方法:用小檗碱与体外培养的人脐静脉内皮细胞(HUVEC)共同孵育,以MTT法检测细胞的增殖活性,免疫细胞化学法检测增殖细胞核抗原(PCNA)的表达,荧光染色法观察凋亡细胞核形态,用Rhodamine123染色,激光共聚焦显微镜检测线粒体膜电位。结果:不同浓度小檗碱与HUVEC共同孵育可明显抑制HUVEC的增殖(P<0.05,P<0.01),呈现一定的浓度依赖性和时效性;20mg/L小檗碱与HUVEC共同孵育48h,可显著降低细胞核PCNA的表达(P<0.01),并可见凋亡细胞数增多、线粒体膜电位明显降低(P<0.01)。结论:小檗碱抑制血管内皮细胞的增殖,并促进血管内皮细胞凋亡,从而抑制肿瘤血管形成,可能是小檗碱抑制肿瘤生长与转移的机制之一。

AIM: To investigate the mechanism of berberine’s inhibiting growth and metastasis of tumor by observing the effects of berberine on proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were incubated with berberine. MTT assay and quantitative immunocytochemistry were used to detect cell proliferation and the expression of proliferation cell nuclear antigen (PCNA). Morphologic changes of apoptosis were observed by fluorescent staining, and Rhodamin123 was used to determinate mitochondrial membrane potential under the laser confocal microscopy. RESULTS: HUVEC proliferation was inhibited by co-incubating with berberine (20 mg/L) for 24 h and berberine (10 mg/L, 20 mg/L) for 48 h (P<0.01, P<0.05,P<0.01). In the presence of berbering (20 mg/L) for 48 h, the expression of PCNA in HUVECs was decreased significantly (P<0.01), typical apoptotic appearance was found and mitochondrial membrane potential declined markedly (P<0.01). CONCLUSIONS: Berberine has a significant inhibitory effect on proliferation of HUVECs and an induced effect on HUVEC apoptosis, which is related with mitochondria. Berberine inhibits the proliferation of vascular endothelial cells to suppress neovascularization, which may be one of the mechanisms underlying inhibitory effect of berberine on growth and metastasis of tumor.