一种肽构建体对静脉曲张患者分化巨噬细胞MMP-2和MMP-9水平的影响。

Effect of a Peptide Construct on Differentiated Macrophage MMP-2 and MMP-9 Levels of Varicose Patients.

作者信息

Esfandiari Golnaz, Biranvand Azim S, Poursaleh Adeleh, Kakavandi Naser, Fattahi Fahimeh, Ghasempour Ghasem, Bahraini Elham, Khosravi Mohsen, Najafi Mohammad

机构信息

Biochemistry Department, Faculty of Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.

Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.

出版信息

Curr Pharm Des. 2019;25(40):4303-4309. doi: 10.2174/1381612825666191029105203.

DOI:10.2174/1381612825666191029105203

PMID:31663473

Abstract

BACKGROUND

The Matrix Metalloproteinase (MMPs) secreted from macrophages can affect the extracellular matrix remodeling process and improve varicose veins.

AIM

The aim of this study was to investigate the MMP-2 and MMP-9 gene expression and activity levels in the differentiated macrophages M2 of subjects with varicose veins, and to evaluate a peptide construct on their catalytic functions.

METHODS

The macrophages were differentiated from the monocytes using M-CSF. The MMP-2 and MMP-9 gene expression and activity levels were measured by RT-qPCR and Zymography techniques, respectively. A peptide construct (ESLCG) was predicted with bioinformatics tools, and was prepared for the study of enzyme functions as compared to Batimastat. Furthermore, the docking studies were obtained for the evaluation of interactions between peptide construct, Batimastat and enzyme 3D structures.

RESULTS

The results showed significant increases in MMP2 and MMP9 gene expression levels (P<0.001 and P<0.004, respectively) and gelatinolytic activities (P<0.001 and P<0.0001, respectively) in the macrophages. In agreement with the inhibitory effects of Batimastat, the peptide construct inhibited the MMP-2 and MMP-9 gelatinolytic activities up to 6.8 and 6.5 folds in the concentration of 150 µM. The docking analyses showed that the Lys187, Arg98, Leu49, Gly189, Leu190, Met97, Tyr53 and Phe57 residues of MMP-2 and the Leu187, His190, Glu402, His401, His405 and His411 residues of MMP-9 are interacted with the atoms of Batimastat and ESLCG peptide.

CONCLUSION

The ESLCG peptide may be applied as an inhibitor of MMP-2 and MMP-9 enzymes in the subjects with varicose veins.

摘要

背景

巨噬细胞分泌的基质金属蛋白酶（MMPs）可影响细胞外基质重塑过程并改善静脉曲张。

目的

本研究旨在调查静脉曲张患者分化的M2巨噬细胞中MMP-2和MMP-9基因表达及活性水平，并评估一种肽构建体对其催化功能的影响。

方法

使用M-CSF将单核细胞分化为巨噬细胞。分别通过RT-qPCR和酶谱技术测量MMP-2和MMP-9基因表达及活性水平。利用生物信息学工具预测一种肽构建体（ESLCG），并制备用于研究其与batimastat相比的酶功能。此外，进行对接研究以评估肽构建体、batimastat与酶三维结构之间的相互作用。

结果

结果显示巨噬细胞中MMP2和MMP9基因表达水平（分别为P<0.001和P<0.004）及明胶酶解活性（分别为P<0.001和P<0.0001）显著增加。与batimastat的抑制作用一致，在150μM浓度下，肽构建体将MMP-2和MMP-9明胶酶解活性分别抑制高达6.8倍和6.5倍。对接分析表明，MMP-2的Lys187、Arg98、Leu49、Gly189、Leu190、Met97、Tyr53和Phe57残基以及MMP-9的Leu187、His190、Glu402、His401、His405和His411残基与batimastat和ESLCG肽的原子相互作用。