雷公藤化合物TW3抗血管生成作用的分子调控机制研究

Molecular regulation mechanism of antiangiogenic effect of Tripterygium wilfordii compound TW3

目的:观察不同剂量雷公藤化合物TW3对人脐静脉内皮细胞(HUVEC)血管内皮生长因子(VEGF)和环氧化酶-2(COX-2)mRNA表达的影响。方法:采用新生儿脐带分离培养出原代HUVEC细胞株,传代培养后制成单细胞悬液,接种培养细胞,将贴壁细胞随机分为模型组(加入佛波酯),TW3低、中、高剂量组(分别加入0.01、0.1、1μg/m L的TW3),阳性对照组(加入佛波酯后及5-Fu注射液),空白组(不作处理),各组置于37℃、5%CO_2培养箱中培养24h,抽提细胞RNA。用RTPCR检测VEGF和COX-2 mRNA的变化,并通过Smart view图像处理系统软件对电泳条带的光密度定量。结果:TW3各组均可明显抑制人脐静脉内皮细胞VEGF和COX-2 mRNA的表达(P<0.05),且存在剂量效应关系。结论:TW3抗血管生成的分子调控机制可能与下调VEGF和COX-2的mRNA表达有关,且随着TW3剂量的增加,下调作用也增强。

Objective： To investigate the effect of various doses of TW3,a compound from Tripterygium wilfordii,on the mRNA expression of vascular endothelial growth factor（ VEGF） and cyclooxygenase-2（ COX-2） in human umbilical vein endothelial cells（ HUVECs）. Methods： The neonatal umbilical cord was used to isolate and culture the primary HUVEC cell line,single cell suspension was prepared after subculture,and the cultured cells were inoculated. The adherent cells were randomly divided into model group（ phorbol ester was added）,low-,medium-,and high-dose TW3 groups（ 0. 01,0. 1,and 1μg/ml of TW3 was added,respectively）,positive control group（ phorbol ester and 5-fluorouracil injection were added）,and blank group（ no treatment）. All the groups were incubated in the incubator with 5% CO_2 at a temperature of 37°C for 24 hours,and then RNA was extracted. Reverse transcription-polymerase chain reaction was used to measure the changes in the mRNA expression of VEGF and COX-2,and Smart View image processing software was applied for the quantification of optical density of electrophoretic band. Results： In all TW3 groups,TW3 significantly inhibited the mRNA expression of VEGF and COX-2（ P〈0. 05）,and there was a dose-effect relationship. Conclusion： The molecular regulation mechanism of antiangiogenic effect of TW3 may be related to the down-regulated mRNA expression of VEGF and COX-2,and down-regulation is enhanced with increasing dose of TW3.