Sheikhbahaei Fatemeh, Khazaei Mozafar, Nematollahi-Mahani Seyed Noureddin
Department of Anatomy, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
Fertility and Infertility Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Adv Pharm Bull. 2018 Mar;8(1):131-139. doi: 10.15171/apb.2018.016. Epub 2018 Mar 18.
Angiogenesis plays an important role in numerous pathophysiological events like cancer. As a result of this, tranilast as an anti-fibrotic drug induces the promising antitumor activities through the inhibition of angiogenesis. Further, Teucrium polium (TP) is a herbal medicine (family Lamaceae) with antitumor properties. This study was conducted to investigate the combination effects of tranilast and T. polium on human umbilical vein endothelial cells (HUVECs) viability and apoptotic genes expression. The HUVECs line was treated using different doses of tranilast and T. polium alone or their combination. The cell cytotoxicity was evaluated using MTT and LDH assays; apoptosis was examined using acridine orange/ethidium bromide staining, nitric oxide (NO) production was evaluated using Griess reaction and the expression of BAX and BCL-2 genes were detected using real-time RT-PCR. One-way analysis of variance (ANOVA) test was used to compare the data in different groups. The survival rate of HUVECs was significantly reduced (p<0.05) in a dose dependent manner by tranilast and T. polium. However, T. polium and tranilast combination significantly (p<0.001) reduced cell viability and increased apoptotic cells as compared to each drug alone. Also, HUVECs treated with Tranilast / T. polium combination showed a reduced level of NO as regards to cells exposed only to Tranilast or T. polium (p<0.05). Furthermore, a significant increase in BAX and a decrease in BCL-2 mRNA expression were observed in combination group (p<0.001). T. polium synergistically increased the antiangiogenic effect of tranilast on in vitro angiogenic model of HUVECs.
血管生成在众多病理生理过程（如癌症）中发挥着重要作用。因此，曲尼司特作为一种抗纤维化药物，通过抑制血管生成诱导出有前景的抗肿瘤活性。此外，皱叶毛建草是一种具有抗肿瘤特性的草药（唇形科）。本研究旨在探讨曲尼司特和皱叶毛建草联合使用对人脐静脉内皮细胞（HUVECs）活力及凋亡相关基因表达的影响。使用不同剂量的曲尼司特和皱叶毛建草单独或联合处理HUVECs细胞系。通过MTT和LDH检测评估细胞毒性；使用吖啶橙/溴化乙锭染色检测细胞凋亡；使用格里斯反应评估一氧化氮（NO）的产生；使用实时RT-PCR检测BAX和BCL-2基因的表达。采用单因素方差分析（ANOVA）检验比较不同组的数据。曲尼司特和皱叶毛建草均以剂量依赖的方式显著降低了HUVECs的存活率（p<0.05）。然而，与单独使用每种药物相比，皱叶毛建草和曲尼司特联合使用显著（p<0.001）降低了细胞活力并增加了凋亡细胞。此外，与仅暴露于曲尼司特或皱叶毛建草的细胞相比，用曲尼司特/皱叶毛建草联合处理的HUVECs中NO水平降低（p<0.05）。此外，联合组中观察到BAX显著增加，BCL-2 mRNA表达显著降低（p<0.001）。皱叶毛建草协同增强了曲尼司特对HUVECs体外血管生成模型的抗血管生成作用。
