莪术醇抑制VEGF诱导的新生血管生成的实验研究

Experimental study of curcumol inhibiting vascular endothelial growth factor-induced neovascularization

摘要:

目的：研究莪术醇对血管内皮生长因子(VEGF)诱导的新生血管生成的作用及机制。方法：体外培养人脐静脉血管内皮细胞，用50ng/mL VEGF和不同浓度莪术醇进行分组处理，采用CCK-8和EdU实验检测细胞增殖，Transwell实验分析细胞迁移能力，管腔形成实验分析内皮细胞血管生成能力，Western blot检测Akt/mTORC1通路变化。结果：CCK-8实验结果显示，400、800μmol/L莪术醇+VEGF组细胞OD450值明显低于VEGF组(均P<0.01)。EdU结果显示，400μmol/L莪术醇+VEGF组细胞增殖率明显低于VEGF组(P<0.001)。Transwell实验和管腔形成实验结果显示，与VEGF组比较，400μmol/L莪术醇+VEGF组迁移细胞数减少，管腔形成的分支数量和分支长度下降(均P<0.001)。Western blot结果显示，莪术醇可显著减少细胞中Akt/mTORC1下游靶点p-Akt和p-S6的表达。结论：莪术醇可抑制VEGF诱导的血管内皮细胞的增殖、迁移和管腔形成，具有强的抑制血管生成的作用，可进一步用于眼底新生血管的治疗研究。

Abstract:

AIM: To study the role and mechanism of curcumol in neovascularization induced by vascular endothelial growth factor(VEGF).METHODS: Human umbilical vein endothelial cells were cultured in vitro and treated with 50ng/mL VEGF and curcumol at different concentrations. Cell proliferation was detected by CCK-8 and EdU assay, the migration ability of cells was analyzed by Transwell assay, the angiogenesis ability of endothelial cells was analyzed by tube formation assay, and the change of Akt/mTORC1 signal pathway was detected by Western blot.RESULTS: CCK-8 results showed that the OD450 value of cells in 400 and 800 μmol/L curcumol+VEGF group was significantly lower than that in VEGF group(all P<0.01). EdU results showed that the rate of cell proliferation in 400 μmol/L curcumol+VEGF group was significantly lower than that in VEGF group(P<0.001). Transwell assay and the formation assay results showed that the number of migratory cells in 400 μmol/L curcumol+VEGF group was decreased, and the number and length of tube branches were also reduced compared with VEGF group(all P<0.001). Western blot results showed that curcumol significantly inhibited the expression of p-Akt and p-S6, which were downstream targets of Akt/mTORC1 pathway in cells.CONCLUSION: Curcumol can inhibit VEGF-induced cell proliferation, migration and tube formation of vein endothelial cells, and has a strong inhibitory effect on angiogenesis, which can be further studied in the treatment of ocular fundus neovascularization.