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利用日本鹌鹑尿囊膜模型通过局部应用瘦素及高、低分子肝素对血管生成进行调节

Modulation of angiogenesis by topical application of leptin and high and low molecular heparin using the Japanese quail chorioallantoic membrane model.

作者信息

Macajova M, Cavarga I, Sykorova M, Valachovic M, Novotna V, Bilcik B

机构信息

Institute of Animal Biochemistry and Genetics, CBs SAS, Bratislava, Slovakia.

St Elizabeth Cancer Institute, Bratislava, Slovakia.

出版信息

Saudi J Biol Sci. 2020 Jun;27(6):1488-1493. doi: 10.1016/j.sjbs.2020.04.013. Epub 2020 Apr 20.

Abstract

Pathological angiogenesis characterized by uncontrollable vessel growth is an accompanying feature of many diseases. The avian embryo chorioallantoic membrane (CAM) is an excellent model for angiogenesis research. In our study we used a less common Japanese quail CAM model for the testing of angiogenic potential of leptin, high-molecular (heparin sodium) andlow-molecular (nadroparin calcium) heparins. Heparins play a significant role in vascular endothelial cell function, and they are able to modulate the activities of angiogenic growth factors. On embryonic day 7 leptin (5 μg per CAM), heparin sodium (75 IU per CAM) and nadroparin calcium (47.5 IU per CAM) in 500 μl PBS were applied on the CAM surface. After 24 h the fractal dimension (Df) of the vasculature was evaluated. Samples from each group were histologically analyzed and and expression were detected by qPCR. Df was significantly increased in the leptin group. A moderate stimulatory effect of heparin sodium and an inhibitory effect of nadroparin calcium were observed. Both leptin and heparin sodium caused a noticeable increase in the CAM thickness compared to the control and nadroparin calcium groups. We observed an increased number of blood vessels and accumulation of fibroblasts. There was no significant impact on gene expression of and 24 h after treatment, however, trends similar to the changes in Df and CAM thickness were present. The resulting effect of nadroparin administration on levels was exactly the opposite to that of leptin ( < 0.05).

摘要

以血管不可控生长为特征的病理性血管生成是许多疾病的伴随特征。鸡胚绒毛尿囊膜(CAM)是血管生成研究的优秀模型。在我们的研究中,我们使用了不太常见的日本鹌鹑CAM模型来测试瘦素、高分子量(肝素钠)和低分子量(那屈肝素钙)肝素的血管生成潜力。肝素在血管内皮细胞功能中发挥重要作用,并且它们能够调节血管生成生长因子的活性。在胚胎第7天,将500μl PBS中含有的瘦素(每CAM 5μg)、肝素钠(每CAM 75IU)和那屈肝素钙(每CAM 47.5IU)应用于CAM表面。24小时后评估脉管系统的分形维数(Df)。对每组样本进行组织学分析,并通过qPCR检测[具体基因名称1]和[具体基因名称2]的表达。瘦素组的Df显著增加。观察到肝素钠有适度的刺激作用,而那屈肝素钙有抑制作用。与对照组和那屈肝素钙组相比,瘦素和肝素钠均导致CAM厚度明显增加。我们观察到血管数量增加和成纤维细胞积聚。治疗24小时后对[具体基因名称1]和[具体基因名称2]的基因表达没有显著影响,然而,存在与Df和CAM厚度变化相似的趋势。那屈肝素给药对[具体指标名称]水平的最终影响与瘦素的影响正好相反(P<0.05)。

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