ABSTRACT

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.

Abbreviations

DMSO: dimethyl sulphoxide; EBP: elastin-binding protein; EDPs: elastin-derived peptides; FBS: foetal bovine serum; Glb1: 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; H₂DCFDA: 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate; DMEM: Dulbecco’s Modified Eagle’s Medium; VVGPGA: Val-Val-Gly-Pro-Gly-Ala.

中文翻译：

---

弹性蛋白衍生肽 VGVAPG 可减少小鼠胚胎成纤维细胞 (3T3-L1) 向脂肪细胞的分化。

摘要

弹性蛋白是存在于结缔组织中的一种高弹性蛋白质。由于蛋白酶活性，弹性蛋白发生水解，在此过程中，弹性蛋白衍生肽 (EDP) 被释放。六肽 VGVAPG 是组成型重复的弹性蛋白和 EDP 构建序列之一。因此，我们研究的目的是确定 VGVAPG 肽对小鼠 3T3-L1 细胞系脂肪生成的影响。3T3-L1 细胞根据先前描述的方案进行分化，并暴露于浓度不断增加的 VGVAPG 或 VVGPGA 肽。获得的结果表明，VGVAPG 肽不会刺激活性氧 (ROS) 产生、c​​aspase-1 激活和 3T3-L1 细胞增殖。在实验的第二部分中，通过油红O染色测量，证明VGVAPG肽减少脂质积累，这通过未分化前脂肪细胞的表达标记物增加的谱图得到证实。在我们的实验中，添加10 nM VGVAPG用于分化为脂肪细胞，与罗格列酮（PPARγ激动剂）治疗组相比，增加了Pref-1、丝氨酸蛋白酶抑制剂E1和脂联素的表达，同时与罗格列酮治疗组相比，降低了VEGF和抵抗素的表达-治疗组。获得的结果表明VGVAPG肽维持3T3细胞处于未分化状态。

缩写

DMSO：二甲基亚砜；EBP：弹性蛋白结合蛋白；EDP​​：弹性蛋白衍生肽；FBS：胎牛血清；Glb1：β-半乳糖苷酶基因；LDL：低密度脂蛋白；PAI-1（丝氨酸蛋白酶抑制剂E1）：纤溶酶原激活剂抑制剂-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: Val-Gly-Val-Ala-Pro-Gly；β-Gal：β-半乳糖苷酶；ORO：油红O；IBMX：3-异丁基-1-甲基黄嘌呤；H ₂ DCFDA：2ʹ,7ʹ-二氯二氢荧光素二乙酸酯；DMEM：Dulbecco 改良 Eagle 培养基；VVGPGA：Val-Val-Gly-Pro-Gly-Ala。