云南重楼中血管内皮生长因子结合因子的活性研究  

Study on activity of VEGF-binding factors in Paris polyphylla

作　　者：李彤彤[1,2] 高飞[2] 段承俐[2] 王向军[2] 钱永常[2,3] 

机构地区：[1]云南农业大学农学与生物技术学院,云南昆明650201 [2]浙江农林大学亚热带森林培育国家重点实验室培育基地,浙江临安311300 [3]美国德克萨斯农工大学综合生物学系

出　　处：《中草药》2015年第15期2251-2256,共6页Chinese Traditional and Herbal Drugs

基　　金：浙江农林大学科研发展基金(2012FR017);浙江省自然科学基金资助项目(LQ14H280007)

摘　　要：目的筛选并分离云南重楼水提物中能与血管内皮生长因子(VEGF)蛋白结合的物质,并探究其对VEGF的刺激作用。方法采用固相化VEGF亲和柱色谱的方法筛选云南重楼水提物中的VEGF结合因子,HPLC法进一步分离纯化该因子成分,并采用MTT法以VEGF敏感型细胞株HepG2为模型验证该因子活性,采用UV、RRLC-QTOF检测,推测该因子可能的成分组成。结果从云南重楼水提物中筛选到能与VEGF蛋白结合的因子(命名为因子B3),HPLC分离纯化后因子B3可以促进VEGF敏感型细胞HepG2的增殖,而对VEGF不敏感型细胞HEK293的增殖没有影响。因子B3和VEGF蛋白对HepG2细胞的增殖促进作用存在叠加效应;而这种效应可以被VEGF抗体阻断;因子B3本身对HepG2细胞的增殖作用也可以被VEGF抗体阻断。通过UV和RRLC-QTOF检测,推测因子B3可能为皂苷类成分。结论云南重楼中得到的因子B3具有促进VEGF功能的活性。Objective To screen and isolate vascular endothelial growth factor（VEGF）-binding factors from Paris polyphylla var.yunnanensis,and explore their property for stimulating VEGF activity.Methods The VEGF-binding factors from water extract of P.polyphylla var.yunnanensis were screened by VEGF-affinity chromatography and further purified by HPLC method.Their activity on proliferation of VEGF-dependent cells was determined by MTT analysis with sensitive cell line HepG2 as model.We also predicted the possible components according to RRLC-QTOF and UV results.Results The VEGF-binding factors screened from the water extract of P.polyphylla var.yunnanensis were named as factor B^3 which included three compounds and could induce the proliferation of VEGF-dependent cell line HepG2 but not the VEGF-independent cell line HEK293.Further studies indicated that factor B^3 had an additive effect with VEGF to induce the proliferation of HepG2,and the additive effect could be attenuated by VEGF antibody.In addition,the proliferation of HepG2 cells induced by factor B^3 alone could also be attenuated by VEGF antibody.Furthermore,based on the results of RRLC-QTOF and UV analysis,we predicted that factor B^3 are probably members of saponin family.Conclusion The factor B^3 isolated from P.polyphylla var.yunnanensis has the property to stimulate VEGF activity.

关 键 词：云南重楼 血管内皮生长因子 因子B^3 亲和柱色谱 皂苷类成分 

分 类 号：R285.5[医药卫生—中药学]