五味子酯甲抑制肝窦内皮细胞功能及血管新生的活性评价

Effect of schisantherin A inhibit liver sinusoid endothelial cell function and action against liver fibrosis relating to angiogenesis

探讨五味子酯甲对肝窦内皮细胞细胞功能及血管新生的影响。0-40μmol·L-1五味子酯甲与SK-HEP-1细胞孵育24 h,MTT法评估五味子酯甲对SK-HEP-1细胞活力的影响。血管内皮生长因子（VEGF）诱导SK-HEP-1增殖,以VEGF受体酪氨酸激酶抑制剂索拉非尼为对照,同时设空白对照组,五味子酯甲与SK-HEP-1细胞孵育,Ed U法检测细胞增殖;荧光探针法检测胞内NO含量和NOS活性;细胞迁移、Matrigel管腔形成实验检测细胞管腔形成;大鼠主动脉环实验检测血管生长变化;转基因斑马鱼实验观察斑马鱼节间血管、功能血管数变化。与空白对照组比较,VEGF诱导SK-HEP-1细胞增殖、NO含量和NOS活性升高;与模型组比较,五味子酯甲显著抑制VEGF诱导的SK-HEP-1细胞增殖、降低NO含量和NOS活性。五味子酯甲能够显著抑制VEGF诱导的SK-HEP-1细胞迁移和管腔形成,抑制大鼠主动脉环血管形成;对转基因斑马鱼节间血管具有显著抑制作用,显著减少斑马鱼功能血管数量。五味子酯甲能够抑制内皮细胞功能,且具有抑制血管新生活性的作用。

To investigate the effect of schisantherin A on liver sinusoid endothelial cell function and angiogenesis. Different dosages（0-40 μmol·L- 1） of schisantherin A were incubated 24 h with SK-HEP-1 cells,and the toxicity of SK-HEP-1 cells was assayed by MTT method. The proliferation of SK-HEP-1 cells were induced by the vascular endothelial growth factor（ VEGF）,with receptor tyrosine kinase inhibitor sorafenib as the control,at the same time,set up the control group,2,20 μmol·L- 1schisantherin A were incubated with SK-HEP-1 cells,cell proliferation was analyzed by Ed U DNA cell proliferation kit. Fluorescence probe method was used to assay the intracellular NO levels and NOS activity. Tube formation was observed using cell migration and a matrigel tube formation assay. Rat aortic ring assay was performed to observe the sprouting vessels from aortic ring. The fluorescence vessels,the number of functional blood vessels,and intersegmental vessel changes of transgenic zebrafish were also observed. Compared with control group,the proliferation of SK-HEP-1 cells induced by VEGF increased and and the level of NO and NOS activity induced; compared with model group,2,20 μmol·L- 1schisantherin A and sorafenib inhibited the proliferation of SK-Hep-1 cells induced by VEGF,and reduced the level of NO and NOS activity. At the dosage of 20 μmol·L- 1,schisantherin A attenuated the migration and tube formation of SK-HEP-1 cells induced by VEGF,and also inhibition the formation of rat aortic rings and intersegmental vessel changes of transgenic zebrafish,and significantly reduce the number of vessels in zebrafish. Schisantherin A has potential effects on function of endothelial cell proliferation and angiogenesis.