A Potential Mechanism for the Anti-Apoptotic Property of Koumine Involving Mitochondrial Pathway in LPS-Mediated RAW 264.7 Macrophages

Abstract

Abstract:Koumine is a kind of alkaloid extracted from Gelsemium elegans (G. elegans). Benth, which has shown promise as an anti-tumor, anxiolytic, and analgesic agent. In our present study, the effect of koumine on lipopolysaccharide (LPS)-mediated RAW 264.7 cell apoptosis was evaluated. MTT assays showed that koumine obviously increased cell viability in LPS-mediated RAW 264.7 macrophages. Preincubation with koumine ameliorated LPS-medicated apoptosis by decreasing reactive oxygen species (ROS) production, which resulted in a significant decrease in the levels of nitric oxide (NO) and inducible nitric oxide synthase (iNOS). In addition, koumine-pretreated RAW 264.7 macrophages exhibited reduction of LPS-induced levels of TNF-α, IL-1β, and IL-6 mRNA. Furthermore, pretreatment with koumine suppressed LPS-mediated p53 activation, loss of mitochondrial membrane potential, caspase-3 activation, decrease of Bcl-2 expression, and elevation of Bax and caspase-3 expressions, suggesting that koumine might act directly on RAW 264.7 cells to inhibit LPS-induced apoptosis. It seems as though the mechanism that koumine possesses is the anti-apoptotic effect mediated by suppressing production of ROS, activation of p53, and mitochondrial apoptotic pathways in RAW 264 cells. Koumine could potentially serve as a protective effect against LPS-induced apoptosis.

Immunomodulatory effects of polysaccharides from purple sweet potato on lipopolysaccharide treated RAW 264.7 macrophages

“…Although apoptosis exists in normal macrophages, it is obvious that LPS treatment significantly augmented apoptotic rate (Figures and ). This result was in accordance with the previous work (Yuan et al, ). According to previous findings, LPS induced apoptosis in macrophages through caspase‐dependent mitochondrial death pathway that decreased mitochondrial membrane potential and promoted activations of caspase‐3 and caspase‐9 (Langford, McGee, Ta, Redens, & Texada, ; Yuan et al, ).…”

“…Zhuang et al () suggested that polysaccharide from Angelica sinensis protected chondrocytes from H 2 O 2 ‐induced apoptosis through its antioxidant effects (Zhuang et al, ). Moreover, RAW264.7 macrophages received koumine treatment exhibited lower LPS‐stimulated apoptosis through suppressing mitochondrial apoptotic pathway (Yuan et al, ). In this study, high apoptosis of macrophages in LPS treatment group was possibly due to the high concentrations of pro‐inflammatory mediators and oxidative damage, while lower apoptosis exsited in polysaccharide treatment groups with lower levels of pro‐inflammatory mediators and higher level of anti‐inflammatory cytokine.…”

Activation of protein kinase C‑α/heme oxygenase‑1 signaling pathway improves mitochondrial dynamics in lipopolysaccharide‑activated NR8383 cells

“…Intracellular ROS levels were detected by using the dichloro-dihydro-fluorescein diacetate (DCFH-DA) probe (Beyotime Institute of Biotechnology, Nanjing, China). In brief, after treatment with LPS for 24 h, macrophages were incubated with 10 µM DCFH-DA at 37˚C for 20 min and then washed twice in PBS (26). DCF fluorescence was monitored with excitation and emission wavelengths of 485 and 535 nm.…”

Ultra-Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS)-Based Pharmacokinetics and Tissue Distribution Study of Koumine and the Detoxification Mechanism of Glycyrrhiza uralensis Fisch on Gelsemium elegans Benth.

“…It was widely used as a classic Chinese herbal medicine to treat malignant skin problems [ 1 ]. Recent years, its anti-tumor [ 2 ], anti-inflammatory [ 3 ], and anxiolytic [ 4 ] activities have also been well studied. On the other side, G. elegans was well-known for its toxicity.…”