Lysozyme Improves the Inhibitory Effects of Panax notoginseng Saponins on Phenotype Transformation of Vascular Smooth Muscle Cells by Binding to Ginsenoside Re.

Yun Huang,Huishan Guo,Hongchao Yang,Rong Wang,Ning Chen,Weiye Shi,Xiaoruo Gan,Lijian Cui,Yan Zhang,Yu Wu,Pin Lv

doi:10.3389/fnut.2021.795888

Abstract

Panax notoginseng saponins (PNS) have been used to treat cardiovascular diseases for hundreds of years in China. Lysozyme can bind to exogenous compounds and promote their activity. Nevertheless, knowledge of whether there is a synergistic role between lysozyme and PNS is far from sufficient. In this study, we show that the mixture of PNS and lysozyme synergistically inhibited platelet derived growth factor BB (PDGF-BB)-induced vascular smooth muscle cell (VSMC) viability, and in the five main components of PNS, GS-Re, but not GS-Rb1, NG-R1, GS-Rg1, or GS-Rd, reduced VSMC viability by combined application with lysozyme. Next, the supramolecular complexes formed by GS-Re and lysozyme were detected by mass spectrometry, and the binding ability increased with the concentration ratio of GS-Re to lysozyme from 4:1 to 12:1. In the supramolecular complexes, the relative contents of α-helix of lysozyme were increased, which was beneficial for stabilizing the structure of lysozyme. The 12:1 mixture of GS-Re and lysozyme (12.8 μmol/L GS-Re+1.067 μmol/L lysozyme) repressed PDGF-BB-induced VSMC viability, proliferation, and migration, which were associated with the upregulated differentiated markers and downregulated dedifferentiated markers. Finally, in CaCl2-induced rodent abdominal aortic aneurysm (AAA) models, we found that the 12:1 mixture of GS-Re and lysozyme slowed down AAA progression and reversed phenotype transformation of VSMCs. Thus, Gs-Re combined with a small amount of lysozyme may provide a novel therapeutic strategy for vascular remodeling-associated cardiovascular diseases.

Highlights

Vascular smooth muscle cells (VSMCs), a kind of highly differentiated cells in normal vessels, are quiescent and possess a contractile phenotype
To screen the main ingredients of Panax notoginseng saponins (PNS) that inhibit VSMC viability through joint application with lysozyme, we determined the effect of the synergism of lysozyme and Ginsenoside Rb1 (GS-Rb1), Notoginsenoside R1 (NG-R1), Ginsenoside Rg1 (GS-Rg1), Ginsenoside Re (GS-Re), or Ginsenoside Rd (GS-Rd), the five main ingredients of PNS, on VSMC viability
Compared with the dimethyl sulfoxide (DMSO) control group, the viability of VSMCs was not changed with the addition of 30 μg/ml lysozyme, GS-Rb1, NG-R1, GS-Rg1, GS-Re, or GS-Rd, and the viability was not affected by the joint application of 15 μg/mL lysozyme and 15 μg/ml GS-Rb1, NGR1, GS-Rg1, or GS-Rd

Summary

Introduction

Vascular smooth muscle cells (VSMCs), a kind of highly differentiated cells in normal vessels, are quiescent and possess a contractile phenotype. As the main component of media, VSMCs provide structural support for vessels, and play a key role in conferring vascular homeostasis. VSMCs are not the terminally differentiated cells, and they can shift from the contractile or differentiated to the synthetic or dedifferentiated phenotype in response to various environmental cues, such as growth factor and mechanical stimulation, manifesting themselves as re-entering the cell cycle and migrating from media into the intima [1, 2], which are the crucial pathological basis of vascular diseases such as atherosclerosis, hypertension, vascular restenosis, arterial aneurysm and diabetic vascular complications [2,3,4]. Agents that can effectively inhibit VSMC proliferation and migration may have a crucial role in the prevention and treatment of vascular remodeling diseases. PNS has numerous pharmacological effects, such as blood dynamics invigoration, cerebral vasodilation, hemostasis, anti-inflammation, antiapoptosis, anti-edema, anti-thromboembolism, anti-coagulation, anti-hyperglycemia, and anti-hyperlipidemia [7, 8]

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Results

Conclusion