从血管内皮生长因子及其受体调控角度探讨地黄梓醇促血管新生作用及分子机制研究

Neovascularization Promotion Effect and Molecular Mechanism of Rehmannia Catalpol in Terms of Regulation of VEGF and Its Receptors

背景血管新生是目前缺血后脑保护研究的新方向之一,地黄主要有效成分梓醇具有促血管新生作用,但其具体作用机制尚未完全明确。目的从血管内皮生长因子(VEGF)及其受体调控角度探讨地黄梓醇促血管新生作用及分子机制。方法实验时间为2019年4—8月,将采用0.9%氯化钠溶液处理的人脐静脉内皮细胞(HUVECs)作为对照组,将采用终浓度为1 mg/ml的梓醇溶液处理的HUVECs作为实验组。采用MTT法检测细胞增殖能力,采用细胞迁移实验检测细胞迁移能力,另分别采用实时荧光定量聚合酶链反应(qRT-PCR)、免疫印迹法检测HUVECs的VEGF、血管内皮生长因子受体1(VEGFR-1)、血管内皮生长因子受体2(VEGFR-2)mRNA及蛋白表达情况。结果(1)对照组HUVECs的OD值低于实验组(P<0.05);与对照组相比,实验组HUVECs相对增殖率为(1.871±0.152)%。(2)以培养0 h为对照,实验组培养12、24 h迁移细胞数量多于对照组(P<0.05);两组HUVECs培养24 h迁移细胞数量均多于培养12 h(P<0.05)。(3)两组HUVECs的VEGFR-1 mRNA及蛋白相对表达量比较,差异无统计学意义(P>0.05);实验组HUVECs的VEGF、VEGFR-2 mRNA及蛋白相对表达量均高于对照组(P<0.05)。结论地黄梓醇通过上调VEGF、VEGFR-2表达而促进HUVECs增殖、迁移,进而发挥促血管新生作用。

Background Neovascularization is one of new research fields for brain protection after ischemia at present,catalpol(as the main component of Rehmannia)has some promotion effect of neovascularization,however its specific mechanism is not yet very clear.Objective To explore the neovascularization promotion effect and molecular mechanism of Rehmannia catalpol in terms of regulation of VEGF and its receptor.Methods This experiment was conducted from April to August 2019.HUVECs treated with 0.9%sodium chloride solution were selected as control group,while HUVECs treated with 1 mg/ml Catalpol solution were selected as test group.MTT method was used to detect the cell proliferative potential,cell migration test was used to detect the cell migration capacity,qRT-PCR and Western blotting method were used to detect the mRNA expressions and protein expressions of VEGF,VEGFR-1 and VEGFR-2.Results(1)OD value of HUVECs in control group was statistically significantly lower than that in test group(P<0.05);compared with that in control group,relative proliferation rate of HUVECs in test group was(1.871±0.152)%.(2)Taking 0 hour after culture as control,number of migrating cells in test group was statistically significantly more than that in control group 12,24 hours after culture(P<0.05);number of migrating cells 24 hours after culture were statistically significantly more than those of 12 hours after culture in the two groups(P<0.05).(3)There was no statistically significant difference in relative mRNA or protein expression quantity of VEGFR-1 between the two groups(P>0.05),while relative mRNA and protein expression quantity of VEGF and VEGFR-2 in test group was statistically significantly higher than that in control group,respectively(P<0.05).Conclusion Rehmannia catalpol can promote the proliferation and migration of HUVECs by up-regulating the expression of VEGF and VEGFR-2,and then play the role of neovascularization promotion.