Department of Lifestyle Disorders and Regenerative Medicine, University of Information Technology and Management in Rzeszow , Rzeszow, Poland.
Adipocyte. 2020 Dec;9(1):234-245. doi: 10.1080/21623945.2020.1770525.
Elastin is a highly elastic protein present in connective tissue. As a result of protease activity, elastin hydrolysis occurs, and during this process, elastin-derived peptides (EDPs) are released. One of the constitutively repeating elastin and EDP building sequences is the hexapeptide VGVAPG. Therefore, the aim of our research was to define the effect of VGVAPG peptide on adipogenesis in a mouse 3T3-L1 cell line. 3T3-L1 cells were differentiated according to a previously described protocol and exposed to increasing concentrations of VGVAPG or VVGPGA peptide. The obtained results showed that VGVAPG peptide does not stimulate reactive oxygen species (ROS) production, caspase-1 activation, and 3T3-L1 cell proliferation. In the second part of the experiments, it was proved that VGVAPG peptide decreased lipid accumulation as measured by oil red O staining, which was confirmed by the profile of increased expression markers of undifferentiated preadipocytes. In our experiments, 10 nM VGVAPG added for differentiating to adipocytes increased the expression of Pref-1, serpin E1, and adiponectin as compared to rosiglitazone (PPARγ agonist)-treated group and simultaneously decreased the expression of VEGF and resistin as compared to the rosiglitazone-treated group. The obtained results show that VGVAPG peptide sustains 3T3 cells in undifferentiated state.
DMSO: dimethyl sulphoxide; EBP: elastin-binding protein; EDPs: elastin-derived peptides; FBS: foetal bovine serum; : gene for beta-galactosidase; LDL: low-density-lipoprotein; PAI-1 (Serpin E1): plasminogen activator inhibitor-1; PBS: phosphate-buffered saline; PPARγ: peroxisome proliferator-activated receptor gamma; Pref-1: preadipocyte factor 1; ROS: reactive oxygen species; VEGF-A: vascular endothelial growth factor-A; VGVAPG: Val-Gly-Val-Ala-Pro-Gly; β-Gal: beta-galactosidase; ORO: oil red O; IBMX: 3-isobutyl-1-methylxanthine; HDCFDA: 2',7'-dichlorodihydrofluorescein diacetate; DMEM: Dulbecco's Modified Eagle's Medium; VVGPGA: Val-Val-Gly-Pro-Gly-Ala.
弹性蛋白是一种存在于结缔组织中的高弹性蛋白。由于蛋白酶的活性，弹性蛋白发生水解，在此过程中，释放出弹性蛋白衍生肽（EDP）。弹性蛋白和 EDP 的一个组成重复序列是六肽 VGVAPG。因此，我们的研究目的是确定 VGVAPG 肽对小鼠 3T3-L1 细胞系脂肪生成的影响。3T3-L1 细胞按照先前描述的方案分化，并暴露于递增浓度的 VGVAPG 或 VVGPGA 肽。获得的结果表明，VGVAPG 肽不会刺激活性氧（ROS）的产生、半胱天冬酶-1 的激活和 3T3-L1 细胞的增殖。在实验的第二部分，证明 VGVAPG 肽减少了脂滴的积累，如油红 O 染色所示，这通过未分化前脂肪细胞标记物表达谱的增加得到证实。在我们的实验中，与罗格列酮（PPARγ 激动剂）处理组相比，在分化为脂肪细胞时添加 10 nM 的 VGVAPG 增加了 Pref-1、丝氨酸蛋白酶抑制剂 E1 和脂联素的表达，同时与罗格列酮处理组相比，降低了 VEGF 和抵抗素的表达。所得结果表明，VGVAPG 肽使 3T3 细胞保持未分化状态。
DMSO：二甲亚砜；EBP：弹性蛋白结合蛋白；EDPs：弹性蛋白衍生肽；FBS：胎牛血清；：β-半乳糖苷酶基因；LDL：低密度脂蛋白；PAI-1（Serpin E1）：纤溶酶原激活物抑制剂-1；PBS：磷酸盐缓冲盐水；PPARγ：过氧化物酶体增殖物激活受体γ；Pref-1：前脂肪细胞因子 1；ROS：活性氧；VEGF-A：血管内皮生长因子 A；VGVAPG：缬氨酰-甘氨酰-缬氨酰-丙氨酰-脯氨酸；β-Gal：β-半乳糖苷酶；ORO：油红 O；IBMX：3-异丁基-1-甲基黄嘌呤；HDCFDA：2'，7'-二氯二氢荧光素二乙酸酯；DMEM：杜尔贝科改良伊格尔培养基；VVGPGA：缬氨酸-缬氨酸-甘氨酸-脯氨酸-甘氨酸-丙氨酸。
