Gentiopicroside (GENT) protects against sepsis induced by lipopolysaccharide (LPS) through the NF-κB signaling pathway

Abstract

Background: Sepsis is a high-mortality disease without effective therapeutic options. The hyperactivation of the monocyte-macrophage system, especially M1 macrophages, triggers the onset of septic shock. Gentiopicroside (GENT), the main active component in the traditional Chinese medicinal herb Radix Gentianae, has been shown to have anti-inflammatory properties. Nevertheless, this anti-inflammatory effect has not been fully elucidated.Methods: In vitro, we stimulated primary bone marrow-derived macrophages (BMMs) or peritoneal elucidated macrophages (PEMs) by lipopolysaccharide (LPS) and interferon (IFN)-γ and pre-treated with GENT and we tested the cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF) α production by enzyme linked immunosorbent assay (ELISA) or real-time quantitative PCR (qPCR). Further, we determined the NF-κB-mediated inflammatory pathway such as IKKα/β and p65 phosphorylation by Western blot. Then we detected the p65 nuclear localization by immunofluorescent staining. Moreover, NF-κB inhibitor and p65-targeted siRNAs were further used to validate the anti-inflammatory mechanism of GENT. In vivo, GENT (50 mg/kg) was administered intragastrically before and after LPS (40 mg/kg) injection. The death time were recorded and the serum levels of IL-1β, IL-6 and TNFα were tested by ELISA, and the IL-1β, IL-6 and TNFα mRNA expression in the lung were test by qPCR and the M1 infiltration in the lung were determined by F4/80 and INOS immunofluorescent staining Results: In vitro, we observed that GENT reduced the inflammatory cytokine production of BMMs stimulated by (LPS)/IFN-γ and ameliorated the phosphorylation of IKKα/β and p65, the degradation of IκBα, and the translocation of p65 into the nucleus. We did not find GENT has any effect on MAPK signaling under LPS/IFN-γ stimulation. NF-κB inhibitor and p65 siRNAs eliminated the inhibition effect of GENT. In vivo, we observed GENT prevented mice from dying in the LPS-induced shock model and decreased the serum levels of IL-1β and IL-6, the mRNA expression of IL-1β, IL-6 and TNFα in lung tissue, and the amount of M1 macrophage infiltration in the lung.Conclusions: GENT prevented LPS/IFN-γ-induced inflammatory cytokine production by macrophages through the NF-κB signaling pathway in vitro and protected against the endotoxin shock induced by LPS in vivo.

NLRC4 gene silencing-dependent blockade of NOD-like receptor pathway inhibits inflammation, reduces proliferation and increases apoptosis of dendritic cells in mice with septic shock

“…A previous study elucidated that NLRP3 and NLRP4 work to regulate production and pyroptosis of IL-18 and IL-1β, thus testifying that NLCR4 silencing causes the inhibition of IL-1β, which is in accordance with our findings [ 31 ]. A prior study also noted that gentiopicroside conferred protection against shock and decreased inflammatory cytokine production of IL-1β, IL-6 and TNF-α in lung tissues [ 32 ]. Given the above key findings, it is reasonable to regard silenced NLRC4 as an important gene when inhibiting inflammatory reactions.…”

Remifentanil repairs cartilage damage and reduces the degradation of cartilage matrix in post-traumatic osteoarthritis, and inhibits IL-1β-induced apoptosis of articular chondrocytes via inhibition of PI3K/AKT/NF-κB phosphorylation

“…Several studies have reported that the expression of MMPs could be changed via the PI3K/AKT pathway (42). NF-κB is a downstream protein of the PI3K/AKT pathway and can regulate the secretion of MMP (20), with inhibition of NF-κB reducing the secretion of MMP (43). The p-PI3K and p-AKT are phosphorylated activated forms of PI3K and AKT (44).…”

“…Activation of the PI3K/AKT pathway increases production of MMPs through a variety of downstream target proteins. Nuclear factor (NF)-κB is one of the target proteins of the PI3K/AKT pathway and is also a key regulator of MMP production as a transcriptional factor (20). The PI3K/AKT pathway is also associated with tumor necrosis factor (TNF)-α-induced activation of OA fibroblast-like synoviocytes, which may be involved in the pathogenesis of OA (21).…”

The Traditional Uses, Phytochemistry, Pharmacology, Toxicology, and Clinical Uses of Metagentiana Rhodantha (Franch.) T.N.Ho and S.W.Liu, an Ethnomedicine in Southwest China

“…Secoiridiods are characteristic substances of MR, while their contents are very low. Studies have indicated that secoiridiod glycosides possess wide biological activities such as anti-inflammatory and immunoregulatory ( Saravanan et al, 2014a ; Saravanan et al, 2014b ; Wang et al, 2019a ), analgesic ( Liu et al, 2016 ; Garrido-Suárez et al, 2020 ), anti-tumor ( Tang et al, 2019 ; Xiao et al, 2020 ) hepatoprotective ( Shao et al, 2017 ; Wu et al, 2017 ), hypoglycemic ( Vaidya et al, 2013a ; Dhanavathy, 2015 ; Vaidya et al, 2012 ; Vaidya et al, 2009 ), neuroprotective ( Wang et al, 2019b ; Wu et al, 2019 ) effects.…”