The inhibitory effect and mechanism of the JZ-VF3 fragment from the venom toxin JZTX-V of Chilobrachys jingzhao on macrophage inflammatory response
Objective To investigate the inhibitory effects of the JZTX-V peptide toxin fragments on inflammatory responses in macro-phages and explore the underlying mechanisms.Methods Western blot was used to examine the effects of three JZTX-V peptide frag-ments on lipopolysaccharide(LPS)-induced inducible nitric oxide synthase(iNOS)expression in RAW 264.7 cells.The nitric oxide(NO)concentration in the cell culture supernatant was measured using the Griess reaction,and the production of superoxide was de-tected by chemiluminescence assay using luminol.Real-time quantitative PCR was employed to measure the expression levels of pro-in-flammatory cytokine mRNA in the cells.Western blot was also used to investigate the effects of JZ-VF3 on the mitogen-activated protein kinase(MAPK)and nuclear factor kappa-B(NF-κB)signaling pathways,and immunofluorescence was used to assess the impact of JZ-VF3 on the nuclear translocation of p65.Results Compared to the LPS-stimulated group,treatment with JZ-VF3 significantly inhibi-ted iNOS activation and the production of NO and superoxide in RAW 264.7 cells.It also suppressed the production of pro-inflammato-ry cytokines in the cells.Western blot and immunofluorescence experiments revealed that JZ-VF3 could inhibit the activation of the MAPK and NF-κB signaling pathways,as well as the nuclear translocation of p65.Conclusion The JZ-VF3 peptide fragment in JZTX-V inhibits inflammatory responses in LPS-induced RAW 264.7 cells by suppressing the activation of the MAPK and NF-κB signaling pathways,demonstrating potential as a peptide anti-inflammatory agent.
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