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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 observe the effects of different doses of tripterygium wilfordii TW3 on the expression of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) mRNA in human umbilical vein endothelial cells (HUVECs). Methods: Primary HUVEC cell lines were isolated and cultured with newborn umbilical cord. After subculture, the cells were made into single cell suspension and inoculated into cultured cells. The adherent cells were randomly divided into model group (adding phorbol ester), TW3 low, High dose group (0.01, 0.1, 1μg / ml respectively), positive control group (after adding phorbol ester and 5-Fu injection) and blank group (without treatment) % CO_2 incubator cultured 24h, extracted cell RNA. The changes of VEGF and COX-2 mRNA were detected by RTPCR and the optical density of electrophoresis bands was quantified by Smart view image processing system software. Results: TW3 groups could significantly inhibit the expression of VEGF and COX-2 mRNA in human umbilical vein endothelial cells (P <0.05), and there was a dose-response relationship. Conclusion: The molecular mechanism of TW3 anti-angiogenesis may be related to the down-regulation of mRNA expression of VEGF and COX-2. With the increase of TW3 dose, the down-regulation effect may be enhanced.