作者：周霞1 ，张文倩 2 ，刘炬 3 ，李丽 1 ，刘付红 3 ，刘炳男 2

单位：

1.250001 山东省济南市，山东中医药大学第二附属医院；2.250014 山东省济南市，山东中医药大学中医学院；3.250014 山东省济南市，山东省千佛山医院；通信作者：周霞，E-mail：lusy2000@126.com

Units：

1.The Second Affiliated Hospital of Shandong Traditional Chinese Medicine University，Jinan 250001，China；2.College of Traditional Chinese Medicine，Shandong Traditional Chinese Medicine University，Jinan 250014，China；3.Shandong Provincial Qianfoshan Hospital，Jinan 250014，China；Corresponding author：ZHOU Xia，E-mail：lusy2000@126.com

关键词：

地黄；梓醇；血管新生；血管内皮生长因子类；药理作用分子作用机制

Keywords：

Rehmannia glutinosa；Catalpinoside；Angiogenesis；Vascular endothelial growth factors；Molecularmechanisms of pharmacological action

CLC：

R 977.9

DOI：

DOI：10.3969/j.issn.1008-5971.2020.01.y04

Funds：

山东省自然科学基金面上项目（ZR2019MH128）；山东省名老中医药专家传承工作室建设项目（鲁中医药函〔2018〕1 号）

摘要：

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。

Abstract：

Background　Neovascularization is one of new research fields for brain protection after ischemia atpresent，catalpol（as the main component of Rehmannia）has some promotion effect of　neovascularization，however itsspecific mechanism is not yet very clear. Objective　To explore the neovascularization promotion effect and molecular mechanismof Rehmannia catalpol in terms of regulation of VEGF and its receptor. Methods　This experiment was conducted from Aprilto August 2019. HUVECs treated with 0.9% sodium chloride solution were selected as control group，while HUVECs treatedwith 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 detectthe mRNA expressions and protein expressions of VEGF，VEGFR-1 and VEGFR-2. Results　（1）OD value of HUVECsin 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，numberof 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 thetwo groups（P<0.05）.（3）There was no statistically significant difference in relative mRNA or protein expression quantity ofVEGFR-1 between the two groups（P>0.05），while relative mRNA and protein expression quantity of VEGF and VEGFR-2in test group was statistically significantly higher than that in control group，respectively（P<0.05）. Conclusion　Rehmanniacatalpol can promote the proliferation and migration of HUVECs by up-regulating the expression of VEGF and VEGFR-2，andthen play the role of neovascularization promotion.

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