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为了探讨肿瘤坏死因子(TNF-α)和雷公藤内酯醇作用Raji细胞VEGF(血管内皮细胞生长因子)的表达及VEGF与ECV血管形成的关系,采用MTT法检测雷公藤内酯醇抑制Raji细胞增殖的影响;用ELISA法对Raji细胞上清的VEGF定量;用基质胶上的内皮细胞网络形成检测ECV304(人类脐静脉内皮细胞起源的细胞系)血管生成;用RT-PCR检测VEGFmRNA含量。结果表明:雷公藤内酯醇对Raji细胞增殖抑制作用呈时间和剂量依赖方式,最适合作用时间为24小时,24小时半数抑制浓度IC50为25nmol/L;Raji细胞上清VEGF的含量在TNF-α组明显高于空白对照组,雷公藤内酯醇处理组则明显低于空白对照组,两者比较有极显著差异(P<0.01);Raji细胞内VEGFmRNA主要为VEGF165和VEGF121,其含量在TNF-α处理组与空白对照组比较有增加,而雷公藤内酯醇抑制VEGFmRNA表达,并呈剂量依赖方式;Raji细胞上清、VEGF(10ng/ml)和TNF-α(10ng/ml)处理的Raji细胞上清在基质胶中作用ECV304细胞后可促进血管形成,而雷公藤内酯醇(25nmol/L)处理的Raji细胞上清和1640培养基作为的对照组加入基质胶中ECV304细胞未见血管形成。结论:在TNF-α、雷公藤内酯醇作用Raji细胞过程中,TNF-α促进VEGF的表达而雷公藤内酯醇则抑制其表达;TNF-α处理的Raji细胞上清在基质胶中促进血管形成,而雷公藤内酯醇则抑制血管形成。
To investigate the expression of vascular endothelial growth factor (VEGF) and the relationship between vascular endothelial growth factor (VEGF) and angiogenesis in Raji cells induced by tumor necrosis factor (TNF-α) and triptolide, the inhibitory effect of triptolide on the proliferation of Raji cells was detected by MTT assay The VEGF level of Raji cell supernatant was quantified by ELISA. Angiogenesis of ECV304 (human umbilical vein endothelial cell origin cell line) was detected by endothelial cell network on Matrigel. VEGF mRNA level was detected by RT-PCR. The results showed that triptolide inhibited the proliferation of Raji cells in a time-and dose-dependent manner. The most suitable time was 24 hours and the IC50 was 25 nmol / L at 24 hours. Group was significantly higher than the control group, the triptolide treatment group was significantly lower than the blank control group, the two were significantly different (P <0.01); VEGF mRNA Raji mainly VEGF165 and VEGF121, the content of TNF- (P <0.05). Compared with the blank control group, triptolide inhibited the expression of VEGFmRNA in a dose-dependent manner. Raji cells supernatant, VEGF (10ng / ml) and TNF-α The ECV304 cells in the supernatant can promote the formation of blood vessels, and triptolide (25nmol / L) treatment Raji cell supernatant and 1640 medium as a control group added to the ECV304 matrigel no vascular formation. Conclusion: In the process of TNF-α and triptolide-induced Raji cells, TNF-α can promote the expression of VEGF and triptolide inhibited the expression of TNF-α. The supernatant of TNF-α-treated Raji cells promoted angiogenesis in Matrigel , While triptolide inhibits angiogenesis.