Department of Public Health, Dietetics and Lifestyle Disorders, Faculty of Medicine, University of Information Technology and Management in Rzeszow, Sucharskiego 2, 35-225, Rzeszow, Poland.
Department of Clinical Biochemistry and Laboratory Diagnostics, Institute of Medicine, University of Opole, Oleska 48, 45-052, Opole, Poland.
Neurochem Res. 2019 May;44(5):1127-1137. doi: 10.1007/s11064-019-02746-z. Epub 2019 Feb 13.
The products of elastin degradation, namely elastin-derived peptides (EDPs), are detectable in the cerebrospinal fluid of healthy individuals and in patients after ischemic stroke, and their number increases with age. Depending on their concentrations, both nitric oxide (NO) and reactive oxygen species (ROS) take part either in myocardial ischemia reperfusion injury or in neurovascular protection after ischemic stroke. The aim of our study was to determine the impact of VGVAPG peptide on ROS and NO production and expression of endothelial nitric oxide synthase (eNos), inducible nitric oxide synthase (iNos) and neuronal nitric oxide synthase (nNos) in mouse cortical astrocytes in vitro. Primary astrocytes were maintained in DMEM/F12 without phenol red supplemented with 10% fetal bovine serum. The cells were exposed to rising VGVAPG peptide concentrations, and ROS and NO production was measured. After 6 h (for mRNA) and 24 (for the protein) of exposure to 10 nM and 1 µM of the peptide, expression of nNos, iNos and eNos was measured. Moreover, the Glb1 siRNA gene knockdown method and Pioglitazone, a peroxisome proliferator-activated receptor gamma (Pparγ) agonist, were applied. Our study shows that the VGVAPG peptide decreased eNos, iNos and nNos mRNA and protein expression in mouse astrocytes in vitro. The VGVAPG peptide also decreased NO production while increasing ROS production in the cells. Furthermore, silencing of the Glb1 gene reversed all effects caused by the VGVAPG peptide. However, due to the lack of sufficient data explaining the molecular mechanism of action of the VGVAPG peptide in the nervous system, more studies in this area are necessary.
弹性蛋白降解产物，即弹性蛋白衍生肽（EDPs），可在健康个体的脑脊液以及缺血性中风患者的脑脊液中检测到，且其数量随年龄增长而增加。取决于其浓度，一氧化氮（NO）和活性氧（ROS）均可参与心肌缺血再灌注损伤，也可参与缺血性中风后的神经血管保护。本研究旨在确定 VGVAPG 肽对体外培养的小鼠皮质星形胶质细胞中 ROS 和 NO 产生以及内皮型一氧化氮合酶（eNOS）、诱导型一氧化氮合酶（iNOS）和神经元型一氧化氮合酶（nNOS）表达的影响。原代星形胶质细胞在不含酚红的 DMEM/F12 中培养，并用 10%胎牛血清进行补充。将细胞暴露于不断增加的 VGVAPG 肽浓度中，并测量 ROS 和 NO 的产生。在肽浓度为 10nM 和 1µM 时暴露于细胞 6 小时（用于 mRNA）和 24 小时（用于蛋白质）后，测量 nNOS、iNOS 和 eNOS 的表达。此外，还应用了 Glb1 siRNA 基因敲低方法和吡格列酮（过氧化物酶体增殖物激活受体γ（Pparγ）激动剂）。本研究表明，VGVAPG 肽可降低体外培养的小鼠星形胶质细胞中 eNOS、iNOS 和 nNOS 的 mRNA 和蛋白质表达。VGVAPG 肽还可降低细胞中 NO 的产生，同时增加 ROS 的产生。此外，Glb1 基因的沉默可逆转 VGVAPG 肽引起的所有作用。然而，由于缺乏充分的数据来解释 VGVAPG 肽在神经系统中的作用分子机制，因此在该领域还需要进行更多的研究。
