CAJ | 학술논문

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TP-LP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）；50 nmol/L的TP和TP-LP作用24 h后，对照组、T P组和 T P-L P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。
목적：비교뢰공등갑소（TP）급기지질체（TP-LP）재체내외대혈관생성적작용，평개지질체작위TP납미재체적가행성。방법채용MTT법측정TP화TP-LP대인제정맥내피세포（HUVECs）증식적영향；채용화흔실험、침습실험、소관형성실험비교이자대HUVECs천이、침습급성관능력적영향；재체내채용기질효전실험비교이자대혈관생성적작용；면역조화검측이자대혈관내피세포생장인자（VEGF）급혈관내피세포점부분자（CD31）적영향。결과 TP화TP-LP균시간、제량의뢰성지억제 HUVECs적증식，TP-LP적억제작용교TP강（P＜0.05）；50 nmol/L적TP화TP-LP작용24 h후，대조조、T P조화 T P-L P조적세포천이거리분별위（246.33±15.82）、（177.00±14.73）、（111.67±17.56）μm ，TP화TP-LP균가억제HUVECs세포적천이（균 P＜0.05），TP-LP적작용교 TP강（P＜0.05）；동대조조비교，TP화TP-LP균가억제HUVECs적침습（균 P＜0.05），TP조화 TP-LP조침습세포수분별위（30.45±2.99）、（11.60±1.53），TP-LP적작용강우TP（P＜0.01）。대조조、TP조화TP-LP조소관수목분별위（54.59±3.75）、（34.51±3.62）、（13.93±2.53），TP화TP-LP가이억제HUVECs소관적생성（균 P＜0.05），TP-LP적작용강우TP（P＜0.05）。체내기질효전실험현시TP-LP억제혈관적형성능력경강（P＜0.01）；면역조화검측적결과현시 TP화TP-LP통과억제VEGF적표체이영향종류신생혈관적생성，CD31표체하강。결론 TP、TP-LP재체외균능명현억제 HUVECs적증식、천이、침습화소관형성，재체내균능통과억제VEGF적표체종이억제종류신생혈관적생성，TP-LP적작용교TP경강대，유망피개발위일충신적항종류혈관생성제제。
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 endothe-lial growth factor (VEGF) and CD31 were 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 50 nmol/L TP and TP-LP for 24 h or 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 su-perior to TP (P<0.05).The in-vivo Matrigel plug experiments showed that TP-LP had stronger inhibitory effect on blood ves-sel formation than TP(P<0.01).Immunohistochemistry showed that TP and TP-LP could suppress the angiogenesis by inhibi-ting the expression of VEGF ,and the expression of CD31 decreased.Conclusion Both TP and TP-LP can inhibit the prolifera-tion ,migration ,invasion and tubule formation of HUVECs in vitro.They can inhibit the angiogenesis by down-regulating the expression of VEGF in vivo.The effects of TP-LP are superior to those of TP ,and TP-LP is expected to be a new anti-angio-genic agent.