Nonpolysaccharide fraction of Lonicerae japonicae Flos attenuates cyclophosphamide-induced immunosuppression associated with modulation of the Keap1/Nrf2/HO-1/GPX4 signalling pathway
In vitro, LJFE dose-dependently activated the Keap1/Nrf2/HO-1 signalling axis and suppressed excessive NO release in LPS-challenged macrophages, and support the potential of LJFE for further development as an immune-enhancing functional food ingredient.
Abstract
Background Cyclophosphamide (CTX)-induced immunosuppression is closely associated with oxidative stress and ferroptosis, but effective therapeutic interventions remain limited. This study investigated the protective effects of the non-polysaccharide fraction of Lonicerae japonicae Flos (LJFE) against CTX-induced immunosuppression and explored its underlying molecular mechanism. Methods LJFE was prepared by water extraction followed by ethanol precipitation, and its chemical profile was characterized using UPLC-MS/MS. Mice with CTX-induced immunosuppression were administered LJFE at doses of 100, 200, and 400 mg/kg for 10 consecutive days. Immune organ indices, serum IgE and IgM levels, oxidative stress markers (SOD, GSH, MDA), and the TBX21/GATA3 ratio were measured. The Keap1/Nrf2/HO-1/GPX4 signalling pathway was investigated via Western blotting and immunofluorescence. Serum metabolomics and molecular docking were applied to screen key differential metabolites and predict binding interactions between characteristic components and target proteins. LPS-stimulated RAW264.7 macrophages were used to verify pathway activation. Results Seventeen chemical constituents were identified in LJFE. In vivo, LJFE markedly restored thymus and spleen indices, increased serum IgE and IgM levels by approximately 45% and 35%, respectively, elevated SOD and GSH activities, reduced MDA accumulation, and normalized the TBX21/GATA3 ratio. Mechanistically, LJFE downregulated Keap1 and upregulated Nrf2, HO-1, and GPX4 expression. Metabolomics analysis identified hydroferulic acid as a key differential biomarker involved in antioxidative regulation. Molecular docking suggested that characteristic components of LJFE exhibit favorable binding affinity toward Keap1 and Nrf2. In vitro, LJFE dose-dependently activated the Keap1/Nrf2/HO-1 signalling axis and suppressed excessive NO release in LPS-challenged macrophages. Conclusion LJFE ameliorates CTX-induced immunosuppression by modulating the Keap1/Nrf2/HO-1/GPX4 pathway, thereby alleviating oxidative stress and ferroptosis. These findings support the potential of LJFE for further development as an immune-enhancing functional food ingredient.
BXGG partially reversed CTX-associated reductions in body weight, thymus and spleen indices and partially restored serum interleukin-2, interleukin-4 (IL-4), and interferon-gamma (IFN-γ) levels, and white blood cell, lymphocyte, platelet, and reticulocyte counts.
This study aimed to evaluate the aggravating effect of overdosed Longdan Xiegan Decoction (LXD) on D-galactosamine-induced liver injury and to investigate the underlying mechanisms. Histopathological examination, serum biochemical analysis (ALT, AST), serum LPS levels, inflammatory cytokine expression, 16S rRNA gene sequencing of gut microbiota, and untargeted fecal metabolomics were performed to assess the effects of LXD at doses of 150, 300, and 600 mg/kg. LC-MS was employed to analyze gut bacterial biotransformation products of LXD. Network pharmacology and molecular docking were used to predict potential genipin-interacting proteins. An LPS-induced LX-2 cell model was established to evaluate the effects of excessive genipin on cell viability and ROS production. Flow cytometry, Western blot, and immunofluorescence were applied to investigate the activation of the TLR4/NF-κB pathway. 600 mg/kg LXD significantly aggravated liver injury and enhanced the gut biotransformation of geniposide to genipin by altering gut microbiota composition. TLR4 was identified as a potential mediator of genipin effects. Moreover, excessive genipin reduced cell viability, increased ROS levels, and elevated expression of TLR4, MyD88, and NF-κB p65, along with promoting nuclear translocation of NF-κB p65 in LX-2 cells. Overdosed LXD aggravates D-galactosamine-induced liver injury through gut microbiota-mediated biotransformation of geniposide to genipin, which subsequently activates the TLR4/NF-κB inflammatory pathway.
Ranran Zhang, Ying Ling, Jiaxin Sun et al.· Fitoterapia· 0 citations
BACKGROUND
This study is the first to elucidate the mechanistic actions of Atriplex portulacoides L. methanolic extract (APME) as a natural treatment of acetic acid-induced ulcerative colitis on rat model.
METHODS
Phytochemical metabolic profiling of APME was determined through the determination of total phenolic and flavonoid contents, alongside negative mode LC-ESI-MS/MS analysis. Furthermore, chromatographic and spectroscopic investigations were employed to fractionate and structurally characterize the metabolites present within APME fractions. The therapeutic potential of APME in UC was evaluated by integrating histological and immunohistochemical analyses. Quantification of oxidative stress, inflammation-related, and lncRNAs (FENDRR and Neat1) gene expression was established.
RESULTS
APME was evaluated for its total phenolic content and total flavonoid content. LC-ESI-MS/MS analysis in negative ionization mode identified 35 bioactive metabolites within APME. Stigmasterol (1) and 20-Hydroxyecdysone (2) were isolated by column chromatography from the n-hexane and ethyl acetate fractions, respectively, and characterized using spectroscopic analysis. APME dose-dependently and significantly (p < 0.05) reduced the pro-inflammatory cytokine TNF-α, mitigated malondialdehyde levels, and restored total antioxidant capacity in colonic tissue compared to the untreated ulcerative group. APME`s treatments markedly upregulated the mRNA expression levels of Nrf2 and HO-1, while concurrently suppressing NF-κB mRNA expression. Also, it showed a regulatory effect on FENDRR and Neat1. Histological, immunohistochemical, and morphometric assessments further supported these findings, demonstrating substantial improvements in colonic architecture and cellular integrity.
DISCUSSION
Notably, APME represents a potential antioxidant and anti-inflammatory therapeutic candidate for UC. It modulates FENDRR and Neat1, revealing promising targets for IBD therapy.
M. Hassan, Marwa M. Mona, Marwa M. Abd-Elsalam et al.· InflammoPharmacology· 0 citations
Background Guazuma ulmifolia is traditionally used for liver disorders, but its protective mechanisms against heavy metal toxicity are poorly defined. This study evaluated the phytochemical profile and hepatoprotective mechanisms of G. ulmifolia butanol extract (Gul-BuOH) against cadmium-induced liver injury. Methods and findings Gul-BuOH was chemically profiled by UPLC-PDA-ESI–qTOF-MS/MS. Cadmium hepatotoxicity was induced in rats, followed by Gul-BuOH treatment (100 and 200 mg/kg). Liver injury, oxidative stress, inflammation, gene expression (Let-7a, HOTAIR), histopathology, HO-1/Sirt-1 immunoreactivity, and serum metabolomic changes were assessed. Chemical profiling led to the annotation of 42 compounds, including mainly flavonoids and phenolic acids, highlighting the rich phytochemical composition of G. ulmifolia. CdCl2 exposure increased hepatic Cd accumulation and elevated ALT, AST, and ALP, reduced TAC, and increased NO and MDA. Gul-BuOH significantly reduced hepatic Cd levels by 2.6- and 3.2-fold, restored TAC by 38.3% and 83.2%, and decreased NO (56.3% and 63.4%) and MDA (45.4% and 54.6%) at 100 and 200 mg/kg, respectively. Inflammatory markers NF-κB-p and TNF-α were markedly suppressed, while miRNA Let-7a was upregulated and lncRNA HOTAIR was downregulated. Histological and immunohistochemical analyses revealed near-complete restoration of hepatic architecture and normalization of HO-1 and Sirt-1 expression at the high dose. Serum metabolomics’ OPLS-DA model performance indicators demonstrated strong reliability, with an R2Y (explained variance) of 0.991 and a Q2 (predictive variance) of 0.988). The model identified 36 significantly altered metabolites that were largely normalized by Gul-BuOH, implicating linoleic acid metabolism, amino acid biosynthesis, and ascorbate-related pathways. Conclusion Gul-BuOH affords dose-dependent protection against Cd-induced liver injury by modulating oxidative stress, inflammation, metal detoxification, and metabolic pathways, supporting the traditional use of G. ulmifolia and its potential as a multi-target hepatoprotective agent.
O. A. Elsabagh, Abdelbaset M. Elgamal, Heba A. Hassan et al.· PLoS ONE· 0 citations
Results showed that SXD attenuated RA progression and reduced inflammatory cytokine levels in rat serum and cell culture supernatants, and support the partial involvement of Pol β-associated processes in the anti-arthritic effects of SXD.
Although microRNAs have been investigated, the reno-protective role of microRNA-205 (miR-205) in cyclophosphamide (CPA)-induced nephrotoxicity has not yet been explored. Therefore, this study evaluated the influence of buspirone on miR-205 and its downstream target EGLN2, focusing on oxidative and ER stress as key drivers of renal injury. This represents a significant gap in nephrotoxicology and epigenetic Toxicology research.
Animals were randomly allocated among four groups (n = 8), including the normal control group and the CPA group that received one intraperitoneal injection of CPA at 200 mg/kg on day 7. Buspirone at 5 or 10 mg/kg per day was administered by oral gavage for ten consecutive days, whereas CPA injected on the seventh day. At experimental completion, blood samples and renal tissues were subjected to analysis using biochemical, histopathological, immunohistochemical, Western blotting, and qRT-PCR techniques.
CPA induced marked renal dysfunction, reflected by elevated SCr, BUN, NGAL, and KIM-1, with evident histopathological damage. This was accompanied by miR-205 suppression, EGLN2 upregulation, impaired Nrf2 signaling, increased PC levels, and stimulation of PERK/eIF2α/ATF4/CHOP-linked ER stress signaling (
p
< 0.01). Concurrently, activation of JNK1, Fos, and NF-κB increased inflammatory cytokines, and initiated caspase-dependent apoptotic signaling (
p
< 0.01). Buspirone pretreatment mitigated these alterations, accompanied by increased miR-205 expression, reduced EGLN2 expression, enhanced Nrf2-associated antioxidant defenses, and attenuation of ER stress, inflammation, and apoptosis, thereby preserving renal function and tissue integrity.
These findings highlight the preventive renoprotective potential of buspirone against CPA-induced nephrotoxicity and underscore the relevance of miR-205/EGLN2 signaling in renal stress responses. Future toxicokinetic and toxicodynamic studies should evaluate buspirone’s safety in tumor treatment.
Ahmed M. El-Dessouki, A. Alrashidi, Mohammad I Jumaa et al.· Frontiers in Pharmacology· 0 citations