Baharara Javad, Amini Elaheh, Musavi Marziyeh
Department of Biology, Research Center For Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, 9183897194, Iran.
Department of Cellular & Molecular Biology, Faculty of Biology, Kharazmi University, Tehran, 14911-15719, Iran.
Iran J Biotechnol. 2017 Sep 27;15(3):179-185. doi: 10.15171/ijb.1208. eCollection 2017.
Bioactive compounds such as terpenoids, chondroitin sulfate, and polysaccharides with added value can be found in prestine marine creatures. These compounds often do have highly valuable therapeutic applications such as being antioxidant, antitumorogenic, anti-inflammatory and anti-angiogenic. For the latter, varieties of angiogenesis factors can suppress this issue within the bodily tissues. The anti-angiogenic and anti-metastatic capacity of a polysaccharide derived from brittle star was investigated. The anti-proliferative effect of derived polysaccharide on umbilical vein endothelial cells (HUVEC) was measured using MTT (dimethyl thiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. The anti-angiogenic effect of the isolated polysaccharide was examined by Chorioallantoic membrane (CAM) assay. The transcriptional expression of VEGF (Vascular Endothelial Growth Factor) was evaluated by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). The anti-metastatic activity was investigated via scratch-wound healing assay. The levels of Paxillin and Matrix Metalloproteinase-9 (MMP-9) expression were analyzed by RT-PCR. Statistical analysis and mean comparisons (< 0.05) were carried out by SPSS 16. Our results elucidated that the brittle star isolated polysaccharide exerted a dose dependent cytotoxic effect on the HUVEC endothelial cells. The CAM assay exhibited potent anti-angiogenic activity . The RT-PCR analysis showed that the extracted polysaccharide (40, 60 µg.mL) down-regulated the VEGF expression. Further, the diminished attachment of endothelial cells demonstrated that the anti-invasiveness of the derived polysaccharide (25, 50 µg.mL) was administrated via down-regulation of paxillin and MMP-9 mRNA expression. Taken together, these results indicated that the polysaccharide extracted from brittle star was able to decrease the viability of the HUVEC cells, to suppress angiogenesis, and possibly act as a natural anti-angiogenic and anti-metastatic marine organic compound against angiogenesis related pathologies.
在原始海洋生物中可以发现具有附加值的生物活性化合物,如萜类化合物、硫酸软骨素和多糖。这些化合物通常具有非常有价值的治疗应用,如抗氧化、抗肿瘤、抗炎和抗血管生成。对于后者,多种血管生成因子可以抑制身体组织内的这个问题。研究了一种来自海蛇尾的多糖的抗血管生成和抗转移能力。使用MTT(3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐)法测定衍生多糖对人脐静脉内皮细胞(HUVEC)的抗增殖作用。通过鸡胚绒毛尿囊膜(CAM)试验检测分离多糖的抗血管生成作用。通过逆转录聚合酶链反应(RT-PCR)评估血管内皮生长因子(VEGF)的转录表达。通过划痕愈合试验研究抗转移活性。通过RT-PCR分析桩蛋白和基质金属蛋白酶-9(MMP-9)的表达水平。使用SPSS 16进行统计分析和均值比较(<0.05)。我们的结果表明,从海蛇尾分离的多糖对HUVEC内皮细胞具有剂量依赖性细胞毒性作用。CAM试验显示出强大的抗血管生成活性。RT-PCR分析表明,提取的多糖(40、60μg/mL)下调了VEGF表达。此外,内皮细胞附着减少表明,衍生多糖(25、50μg/mL)通过下调桩蛋白和MMP-9 mRNA表达发挥抗侵袭作用。综上所述,这些结果表明,从海蛇尾提取的多糖能够降低HUVEC细胞的活力,抑制血管生成,并可能作为一种天然的抗血管生成和抗转移海洋有机化合物,用于对抗与血管生成相关的疾病。