查看更多>>摘要：It has been recognized that peptidoglycan recognition proteins (PGRPs), structurally conserved molecules, play crucial roles in the innate immunity of invertebrate. However, few studies have been taken to explore their potential functions. In this study, a novel PGRP from the razor clam Sinonovacula constrict designated as ScPGRPS6 was identified and characterized. The open reading frame (ORF) of ScPGRP-S6 was 666 bp in length, encoding a protein of 221 amino acid with a signal peptide (1-30) and a typical PGRP domain (39-187). The sequence alignment combined with phylogenetic analysis collectively confirmed that ScPGRP-S6 was a novel member belonging to PGRP-S family. The mRNA transcript of ScPGRP-S6 in the hepatopancreases was significantly upregulated after peptidoglycan (PGN) stimulation, while it was moderately up-regulated after lipopolysaccharide (LPS) stimulation. The result of immunofluorescence detection demonstrated that the positive signal enhanced obviously after Vibrio parahaemolyticus challenge. Notably, the recombinant protein of ScPGRP-S6 (designed as rScPGRP-S6) exhibited high agglutination activity towards V. parahaemolyticus but weak to Staphylococcus aureus. Furthermore, rScPGRP-S6 showed strong amidase and antibacterial activity in the presence of Zn2+. Collectively, our results manifested that ScPGRP-S6 could act as a scavenger in the innate immune response of S. constricta.
查看更多>>摘要：Acute hypoxic stress can lead to immune response in fish, but the molecular mechanism of muscle immunity in fish under acute hypoxia are still unclear. In this study, we carried out the effect of signal transducer and activator of transcription3(STAT3) and vascular endothelial growth factor A(VEGFA) on muscle immune responses of Japanese flounder (Paralichthys olivaceus) during acute hypoxic stimulation (1.65 +/- 0.28mg/L O-2; 3h, 6h, 12h, 24h) and reoxygenation (7.30 +/- 0.40mg/L O-2; R12h, R24h, R48h). In situ hybridization (ISH) showed that STAT3 and VEGFA RNA were co-located in the skeletal muscle of Japanese flounder. Japanese flounder was seriously affected by hypoxia for 3h and 6h. The expression of STAT3 and VEGFA increased significantly. The methylation levels of STAT3 5'UTR region and VEGFA promoter region were significantly lower than those in normoxia group, which was negatively correlated with the expression levels of STAT3 and VEGFA. The enzyme activities (LDH, ALT, AST, ALP) changed significantly. In addition, enzyme-linked immunosorbent assay (ELISA) detected a positive correlation between serum VEGFA concentration and muscle VEGFA mRNA. The current study have shown that Japanese flounder responded to acute hypoxic stress at multiple metabolic levels by changing DNA methylation status and activating transcription factors such as HIF-1 alpha, Nrf2 and STAT3. It is significant for the scientific development of aquaculture through analyzing the effects of hypoxia on biological immunity.
查看更多>>摘要：Matrix metalloproteinases (MMPs) are highly expressed in leukocytes and macrophages, which play a role in the innate immune response. Here, the cDNA sequence of MMP25 from Japanese sea bass (Lateolabrax japonicus) (LjMMP25) was identified. Phylogenetic analysis revealed that LjMMP25 was most closely related to large yellow croaker MMP25. Multiple sequence alignment of LjMMP25 with MMP25 sequences from other teleosts revealed that regions of known functional importance were highly conserved. Expression analysis revealed that LjMMP25 was highly expressed in the head kidney and widely expressed in other tissues including gill, spleen, and liver. LjMMP25 was found to regulate inflammatory cytokine production and promote phagocytosis and bacterial killing in monocytes/macrophages (MO/M?). Furthermore, LjMMP25 regulated the inflammatory response by modulating NF-kappa B signaling. These findings reveal new information about the role of LjMMP25 in regulating pro inflammatory responses in this species.
NSTL
Elsevier