雷公藤甲素及其脂质体对血管生成的抑制作用  

Inhibitory Effects of Triptolide and Triptolide-loaded Liposome on Angiogenesis

作　　者：杨锐[1] 李红波[1] 王兵[1] 邹声泉[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院普通外科,武汉430030

出　　处：《华中科技大学学报（医学版）》2014年第2期137-141,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家高技术研究发展计划(863计划)资助项目(No.2010DFA31870)

摘　　要：目的比较雷公藤甲素(TP)及其脂质体(TP-LP)在体内外对血管生成的作用,评价脂质体作为TP纳米载体的可行性。方法采用MTT法测定TP和TP-LP对人脐静脉内皮细胞(HUVECs)增殖的影响;采用划痕实验、侵袭实验、小管形成实验比较二者对HUVECs迁移、侵袭及成管能力的影响;在体内采用基质胶栓实验比较二者对血管生成的作用;免疫组化检测二者对血管内皮细胞生长因子(VEGF)及血管内皮细胞粘附分子(CD31)的影响。结果 TP和TPLP均时间、剂量依赖性地抑制HUVECs的增殖,TP-LP的抑制作用较TP强(P<0.05);50nmol/L的TP和TP-LP作用24h后,对照组、TP组和TP-LP组的细胞迁移距离分别为(246.33±15.82)、(177.00±14.73)、(111.67±17.56)μm,TP和TP-LP均可抑制HUVECs细胞的迁移(均P<0.05),TP-LP的作用较TP强(P<0.05);同对照组比较,TP和TP-LP均可抑制HUVECs的侵袭(均P<0.05),TP组和TP-LP组侵袭细胞数分别为(30.45±2.99)、(11.60±1.53),TP-LP的作用强于TP(P<0.01)。对照组、TP组和TP-LP组小管数目分别为(54.59±3.75)、(34.51±3.62)、(13.93±2.53),TP和TP-LP可以抑制HUVECs小管的生成(均P<0.05),TP-LP的作用强于TP(P<0.05)。体内基质胶栓实验显示TP-LP抑制血管的形成能力更强(P<0.01);免疫组化检测的结果显示TP和TP-LP通过抑制VEGF的表达而影响肿瘤新生血管的生成,CD31表达下降。结论 TP、TP-LP在体外均能明显抑制HUVECs的增殖、迁移、侵袭和小管形成,在体内均能通过抑制VEGF的表达从而抑制肿瘤新生血管的生成,TP-LP的作用较TP更强大,有望被开发为一种新的抗肿瘤血管生成制剂。Objective To compare the effect of triptolide（TP）and triptolide-loaded liposome（TP-LP）on angiogenesis in vitro and in vivo in order to examine the feasibility of nano liposome as TP carrier.Methods The effects of TP and TP-LP on the proliferation of human umbilical vein endothelial cells（HUVECs）were detected by MTT.The migration,invasion and tube formation ability of HUVECs were determined by scratch assay,invasion assay and tube formation assay,respectively.The effects of TP and TP-LP on angiogenesis in vivo were examined by Matrigel plug assay,and the expression of vascular endothelial growth factor（VEGF）and CD31were immunohistochemically detected.Results TP and TP-LP time-and dose-dependently inhibited the proliferation of HUVECs.The inhibitory effect of TP-LP was superior to that of TP（P〈0.05）.The migration distance of HUVECs treated with 50nmol/L TP and TP-LP for 24hor not was（246.33±15.82）μm,（177.00±14.73）μm and（111.67±17.56）μm in control,TP and TP-LP groups.Both TP and TP-LP could inhibit the migration of HUVECs（P〈0.05）,and the inhibitory effect of TP-LP was stronger than that of TP（P〈0.05）.The invasion of HUVECs was suppressed in TP and TP-LP groups compared with control group（P〈0.05）,and the number of invasive cells was（30.45±2.99）and（11.60±1.53）in TP and TP-LP groups,respectively,which indicated that TP-LP was superior to TP in terms of the effect on HUVECs invasion（P〈0.01）.The number of tubules was（54.59±3.75）,（34.51±3.62）and（13.93±2.53）in control,TP and TP-LP groups respectively.TP and TP-LP could inhibit the formation of HUVECs tubules（P〈0.05）,and TP-LP was superior to TP（P〈0.05）.The in-vivo Matrigel plug experiments showed that TP-LP had stronger inhibitory effect on blood vessel formation than TP（P〈0.01）.Immunohistochemistry showed that TP and TP-LP could suppress the angiogenesis by inhibiting the expression of VEGF,and the expression of CD31decreased.Conclusion Both TP and TP-LP can inhibit

关 键 词：雷公藤甲素 脂质体 细胞增殖 血管生成 

分 类 号：R730.52[医药卫生—肿瘤]