Berberine Regulated miR150-5p to Inhibit P2X7 Receptor, EMMPRIN and MMP-9 Expression in oxLDL Induced Macrophages.

Lin Lu,Xia Xue,Ting Wang,Jianjian Huang,Jianmin Li,Zhouqing Huang

doi:10.3389/fphar.2021.639558

Abstract

Elevated extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase-9 (MMP-9) in oxidized low density lipoprotein (oxLDL)-induced macrophages leads to the progression of vulnerable plaques by degradation of the extracellular matrix. Our previous report showed that berberine regulates the expression of both EMMPRIN and MMP-9. In addition, P2X7 receptor (P2X7R) upregulation plays a crucial role in the development of atherosclerosis. However, it is unclear whether berberine regulated P2X7R level to inhibit both EMMPRIN and MMP-9 expession in macrophages. In the present study, we investigated the impact of berberine on P2X7R expression and the regulation of P2X7R in the expression of EMMPRIN and MMP-9 in oxLDL-induced macrophages. We found that P2X7R expression was increased, miR150-5p was reduced in oxLDL-induced macrophages, relatively. And A-438079 (a P2X7R inhibitor) or miR150-5p mimic treatment greatly reversed the upregulation of EMMPRIN and MMP-9 expression. Moreover, A-438079 significantly reduced oxLDL-induced AMP-activated protein kinase-α (AMPK-α) phosphorylation and reversed the activation of mitogen-activated protein kinase (MAPK), which in turn decreased the expression of EMMPRIN and MMP-9. These findings illustrate that P2X7R suppresses EMMPRIN and MMP-9 expression by inhibiting the AMPK-α/MAPK pathway in oxLDL-induced macrophages. Accordingly, exposure to berberine markedly upregulated miR150-5p, decreased P2X7R expression and downregulated MMP-9 and EMMPRIN levels in oxLDL-induced macrophages, resulting in AMPK-α/MAPK (JNK, p38, and ERK) inactivation. Overall, these results indicate that berberine increased miR150-5p level, subsequently inhibits P2X7R-mediated EMMPRIN and MMP-9 expression by suppressing AMPK-α and MAPK signaling in oxLDL-induced macrophages.

Highlights

Atherosclerosis is a chronic and progressive inflammatory disease and is closely correlated with oxidized low density lipoprotein (oxLDL)-activated macrophages, which release many inflammatory factors that affect atherosclerotic plaque stability (Galis et al, 1995; Di Pietro et al, 2016; Tabas and Bornfeldt, 2016)
We showed that extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase-9 (MMP-9) expression obviously increased after stimulation with oxLDL, and suppression of P2X7 receptor (P2X7R) expression by A-438079 significantly inhibited the ox-LDL-induced upregulation of MMP-9 and EMMPRIN (Figures 1C,H), and their relative mRNA level represented the same tendency (Figures 1F,G)
MiR-150-5p overexpression directly inhibited all of P2X7R, EMMPRIN, and MMP-9, suggesting that berberine inhibited P2X7R by upregulating miR-150-5p to reduce EMMPRIN and MMP9 expression in oxLDL-stimulated macrophages

Summary

Introduction

Atherosclerosis is a chronic and progressive inflammatory disease and is closely correlated with oxLDL-activated macrophages, which release many inflammatory factors that affect atherosclerotic plaque stability (Galis et al, 1995; Di Pietro et al, 2016; Tabas and Bornfeldt, 2016). Mounting evidence has suggested that elevated EMMPRIN and MMP-9 expression in atherosclerotic plaques (Schmidt et al, 2006) or in atherosclerosis-related cells (such as oxLDLstimulated monocytes/macrophages and coronary smooth muscle cells) (Major et al, 2002) contributes to extracellular matrix degradation and promotes plaque rupture (Yoon et al, 2005; Joghetaei et al, 2013), which indicates that EMMPRIN and MMP-9 play important regulatory roles in the progression of atherosclerosis. P2X7R is expressed in endothelial cells and macrophages, which infiltrate the atherosclerotic plaques in human carotid arteries (Burnstock and Knight, 2004). Whether P2X7R regulates EMMPRIN and MMP-9 expression in oxLDL-stimulated macrophages remains unclear

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Conclusion