[关键词]
[摘要]
目的 探讨丹参水溶性成分对肝窦内皮细胞功能及血管新生的影响。方法 采用肝窦状内皮细胞HHSEC 细胞株,与不同浓度丹参水溶性成分丹酚酸A、丹酚酸C、丹参素钠、咖啡酸、迷迭香酸、丹酚酸B 共孵育24 h,CCK-8 法评估各成分细胞毒性。采用内皮细胞生长补充因子(ECGS)诱导HHSEC 细胞增殖,以索拉非尼为阳性对照,EdU 法检测细胞增殖;荧光探针法检测胞内NO 水平和NOS 活力;免疫荧光法检测CD31 表达;转基因斑马鱼实验观察斑马鱼荧光血管数量;鸡胚绒毛尿囊膜实验检测血管面积。结果 与对照组比较,ECGS 能够诱导HHSEC 细胞增殖,NO 水平和NOS 活性升高,CD31 表达升高;与模型组比较,索拉非尼及丹酚酸C、丹酚酸B、丹酚酸A 能显著抑制ECGS 诱导的HHSEC 细胞增殖,降低细胞NO 量和NOS 活性,CD31 表达明显降低。丹酚酸B、丹酚酸A、咖啡酸能够显著抑制鸡胚绒毛尿囊膜血管新生;丹酚酸C 对转基因斑马鱼功能血管数量具有显著抑制作用。结论 丹参水溶性成分丹酚酸A、B、C 能够抑制肝窦内皮细胞功能,且具有抑制血管新生的活性。
[Key word]
[Abstract]
Objective To investigate the effect of water-soluble components from Salvia miltiorrhiza on liver sinusoid endothelial cell function and angiogenesis. Methods Different dosages of water-soluble components from S. miltiorrhiza were incubated for 24 h with HHSEC cell line, and the toxicity of HHSEC cells was assayed by CCK-8 method. The proliferation of HHSEC cells was induced by endothelial cell growth supplements (ECGS), with receptor tyrosine kinase inhibitor sorafenib as positive control, cell proliferation was analyzed by EdU DNA cell proliferation kit; Fluorescence probe method was used to assay the intracellular NO level and NOS activity; Immunofluorescence method was performed to observe the expression of CD31; The transgenic zebrafishes were incubated for 24 h with each component. The fluorescence vessels, the number of functional blood vessels, and intersegmental vessel changes were observed; Chicken chorioallantoic membranes were incubated for 24 h with each component, Image Analysis Software was used to analyze the vessel area changes. Results Compared with control group, the proliferation of HHSEC cells induced by ECGS increased, the level of NO and NOS activity reduced and the expression of CD31 increased; Compared with the model group, salvianolic acid A, salvianolic acid B, salvianolic acid C, and sorafenib inhibited the proliferation of HHSEC cells induced by ECGS, reduced the level of NO, NOS activity, and expression of CD31. The vessel area of chicken chorioallantoic membranes was decreased in sorafenib, salvianolic acid A, salvianolic acid B and caffeic acid. Salvianolic acid C also significantly inhibited the intersegmental vessel changes of transgenic zebrafish. Conclusion Salvianolic acid A, salvianolic acid B, and salvianolic acid C have the potential effects on function of endothelial cell proliferation and angiogenesis.
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[基金项目]
重大科技专项(2014ZX10005001);国家自然科学基金资助项目(81173405,81473404,81270053)