Bupleurum chinense Polysaccharide Improves LPS-Induced Senescence of RAW264.7 Cells by Regulating the NF-κB Signaling Pathway.

Mengran Xu,Tan Li,Ye Shen,Junhong Ge,Shengyang Sun,Xin Sun,Ademar A Da Silva Filho

doi:10.1155/2020/7060812

Abstract

Macrophages are important inflammatory cells that play a vital role in inflamm-aging. Bupleurum chinense polysaccharide (BCP), an effective component of the Bupleurum chinense herb, exerts multiple beneficial pharmacological effects, such as improving immunity and antioxidant activity. However, the effects of BCP on macrophage-aging and inflamm-aging are yet to be established. In this study, we examined the effects of BCP on proliferation, inflammatory cytokines, β-galactosidase (SA-β-gal), senescence-associated heterochromatin foci (SAHF), reactive oxygen species (ROS), mitochondrial membrane potential, p53, p16, and p65/NF-κB signaling proteins in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. BCP significantly inhibited production of interleukin-1α (IL-1α), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), reduced the expression of SA-β-gal and formation of SAHF, as well as ROS level, and stabilized the mitochondrial membrane potential in RAW264.7 cells stimulated with LPS. Furthermore, BCP inhibited the expression of aging-related genes, p53 and p16, suppressed phosphorylation of p65 protein, and enhanced the expression of I-κBα protein through the NF-κB signaling pathway in LPS-stimulated RAW264.7 cells. Accordingly, we conclude that BCP effectively suppresses inflamm-aging by reducing inflammatory cytokine levels and oxidative stress production following activation of the NF-κB signaling pathway in RAW264.7 cells stimulated with LPS. Our collective findings support the utility of BCP as a novel pharmaceutical agent with potential anti-inflamm-aging effects.

Highlights

Macrophages are important inflammatory cells that play a vital role in inflamm-aging
We examined the effects of Bupleurum chinense polysaccharide (BCP) on proliferation, inflammatory cytokines, β-galactosidase (SA-β-gal), senescenceassociated heterochromatin foci (SAHF), reactive oxygen species (ROS), mitochondrial membrane potential, p53, p16, and p65/ NF-κB signaling proteins in lipopolysaccharide (LPS)-stimulated RAW264.7 cells
Missouri, USA), RPMI1640 medium and fetal bovine serum from Gibco Company (California, USA), enzyme linked immunosorbent assay (ELISA) kits were purchased from ABclonal Company (Wuhan, China), and MTT, NP40 protein lysis buffer, penicillin, streptomycin, BCA protein assay, cell senescence β-galactosidase staining, reactive oxygen species assay, and mitochondrial membrane potential assay kits were acquired from Biyuntian Company (Shanghai, China)

Summary

Introduction

Macrophages are important inflammatory cells that play a vital role in inflamm-aging. We examined the effects of BCP on proliferation, inflammatory cytokines, β-galactosidase (SA-β-gal), senescenceassociated heterochromatin foci (SAHF), reactive oxygen species (ROS), mitochondrial membrane potential, p53, p16, and p65/ NF-κB signaling proteins in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. We conclude that BCP effectively suppresses inflamm-aging by reducing inflammatory cytokine levels and oxidative stress production following activation of the NF-κB signaling pathway in RAW264.7 cells stimulated with LPS. Polysaccharides are natural biological macromolecular compounds composed of monosaccharides Due to their lack of side-effects on normal cells in vivo, polysaccharides are known as “a biological reaction regulators” and represent an important active component of traditional Chinese medicines (TCMs) [19,20,21]. Our findings provide a theoretical basis for further research on the anti-inflamm-aging effect of BCP

Methods

Results

Conclusion