To understand the progression of metaflammation,elucidating the age-dependent role of SOCS8 deficiency in socs8^(-/-)zebrafish is crucial,as this may serve as a disease model for metabolic disorders because of the gra...To understand the progression of metaflammation,elucidating the age-dependent role of SOCS8 deficiency in socs8^(-/-)zebrafish is crucial,as this may serve as a disease model for metabolic disorders because of the gradual dysregulation of immunity,metabolism,and the gut microbiota observed in this species.Thus,both juvenile(1-month-old)and adult(3-month-old)mutant socs8^(-/-)as well as wild-type zebrafish were tested for changes in growth,histopathology,transcriptomes,and microbiota composition.Histological analysis revealed a marked increase in oil droplets and a reduction in hepatocyte nuclear density for the comparison between socs8^(-/-)and wild type fish,indicative of metabolic dysfunction-associated fatty liver disease(MAFLD).The liver in 1-month-old(1 mo)socs8^(-/-)zebrafish had changes in“arginine and proline metabolism”,alongside an increased per-centage of M0 and M2 macrophages.This suggests a protective response from macrophages,which is consistent with the GO term of“negative regulation of leukocyte differentiation”.The enhanced fatty acid metabolism in 1 mo may serve as an indicator of free fatty acid-induced hepatotoxicity associated with simple steatosis but not with liver dysfunction.These findings aligned with the higher growth of 1 mo socs8^(-/-)fish.In 3-month-old(3 mo)socs8^(-/-)fish,the GO terms“steroid hormone biosynthesis”and“steroid metabolic process”indicated the presence of endoplasmic reticulum stress,which contributes to the development of steatohepatitis.DNA repair pathways and associated terms indicated hepatocyte death.The presence of increased dendritic cells,along with expanded activated dendritic cells in nonalcoholic steatohepatitis(NASH),indicated that the 3 mo socs8^(-/-)model may represent the late stage of MAFLD.In the gut,imaging results from larvae and single-cell sequencing of intestinal mucosa cells from 3 mo socs8^(-/-)fish revealed a consistent accumulation of lymphocytes,with the number of innate immune cells higher in larvae but lower in adult fish,compared to wide type fish.In the 1 mo socs8^(-/-)fish,enhanced mucin secretion,suggested by a greater number of goblet cells and upregulated“SNARE interactions in vesicular transport”,was in line with“mannose type O-glycan biosynthesis”.The same group had upregulated IL-13 and its receptors,aligning with the GO term“wound healing”and the idea that innate lym-phocytes producing IL-13 may function as regulators of immunity and tissue remodeling.The enriched pathways“arachidonic acid metabolism”and“intestinal immune network for IgA production”at 1 month,indicating enhanced proliferation of intestinal epithelial cells and improved host‒microbe interactions,respectively.Comparing 1 mo socs8^(-/-)with wide type fish,higher levels of Prevotella indicate enhanced glucose metabolism due to their role in polysaccharide breakdown.Meanwhile,lower levels of Veillonella indicate oxidative stress,potentially associated with a reduced lumen at one month.Intestinal bacterial overgrowth in 3 mo socs8^(-/-)fish(compared to 1 mo socs8^(-/-))contributes to the exacerbation of liver dysfunction.Compared 3 mo socs8^(-/-)with wide type fish,despite an increase in T/NK cells,elevated fatty acid metabolism may impair the function of intestinal lymphocytes.Therefore,the symptoms observed in the gut‒liver axis suggest that socs8^(-/-)zebrafish may serve as a model for MAFLD.展开更多
This study examined the effects of Picria fel-terrae Lour.(PFL)on soybean meal-induced enteritis in zebrafish(Danio rerio)and evaluated whether its increased resistance to Aeromonas hydrophila using both in vivo and i...This study examined the effects of Picria fel-terrae Lour.(PFL)on soybean meal-induced enteritis in zebrafish(Danio rerio)and evaluated whether its increased resistance to Aeromonas hydrophila using both in vivo and in vitro approaches.Compared to plain soybean meal,the addition of 0.05%,0.1%,or 0.2%PFL to zebrafish larval diet reduced intestinal neutrophil(0.1%and 0.2%PFL),macrophage(0.05%and 0.1%PFL),and T cell aggregation(0.01%,0.05%and 0.1%PFL).Following a six-week feeding trial with 0.1%PFL,the hindgut of 3-month-old zebrafish showed a higher length/width ratio of intestinal ridges and the regulation of macrophages and CD4+cells compared to fish fed soybean meal.Immune cell infiltration analysis also demonstrated immune modulatory effects of PFL,such as decreased activated memory CD4 T cells and increased M2 macrophages,and we observed a reduction in nuclear density in the liver,accompanied by an increase in PAS-stained glycogen and p-STAT3 signals.Enrichment analyses revealed that PFL may mitigate intestinal inflammation by upregulating genes involved in DNA replication and downregulating inflammatory processes,such as leukocyte chemotaxis,lysosomes,and phagosomes.The enriched hepatic terms and pathways indicated that PFL facilitated the metabolism of fatty acids and monocarboxylic acids while inhibiting the alteration of tight junctions and the generation of reactive oxygen species by NADPH oxidases.Hexanorcucurbitacin F and its target genes were revealed by network pharmacological analysis and molecular docking,and its target proteins Src and Ptk2ab were related to macrophage function.PFL increased Bifidobacterium longum and species of Faecalitalea,which may contribute to gut homeostasis.During the Aeromonas hydrophila challenge,PFL resulted in a higher survival rate and reduced swelling of the intrabronchial lymphoid tissue compared to soybean meal.PFL water extract also inhibited the growth of A.hydrophila in vitro.In conclusion,PFL dietary inclusion restored intestinal im-mune homeostasis by modulating the function and aggregation of immune cells,stimulating cell regeneration,rebalancing gut flora,and controlling hepatic metaflammation.These findings demonstrate that PFL enhances immune responses across the gut-liver axis and improves mucosal antibacterial defenses in fish,and is a potential functional feed additive.展开更多
基金funded by the Strategic Priority Research Program of Chinese Academy of Sciences(XDA24010206)as well as the Supporting Project for Developing Industrial Science and Technology,Jiangxi Innovation and Incubation Center of Industrial Technologies,Chinese Academy of Sciences(ZKYJXZXCY-202210).
文摘To understand the progression of metaflammation,elucidating the age-dependent role of SOCS8 deficiency in socs8^(-/-)zebrafish is crucial,as this may serve as a disease model for metabolic disorders because of the gradual dysregulation of immunity,metabolism,and the gut microbiota observed in this species.Thus,both juvenile(1-month-old)and adult(3-month-old)mutant socs8^(-/-)as well as wild-type zebrafish were tested for changes in growth,histopathology,transcriptomes,and microbiota composition.Histological analysis revealed a marked increase in oil droplets and a reduction in hepatocyte nuclear density for the comparison between socs8^(-/-)and wild type fish,indicative of metabolic dysfunction-associated fatty liver disease(MAFLD).The liver in 1-month-old(1 mo)socs8^(-/-)zebrafish had changes in“arginine and proline metabolism”,alongside an increased per-centage of M0 and M2 macrophages.This suggests a protective response from macrophages,which is consistent with the GO term of“negative regulation of leukocyte differentiation”.The enhanced fatty acid metabolism in 1 mo may serve as an indicator of free fatty acid-induced hepatotoxicity associated with simple steatosis but not with liver dysfunction.These findings aligned with the higher growth of 1 mo socs8^(-/-)fish.In 3-month-old(3 mo)socs8^(-/-)fish,the GO terms“steroid hormone biosynthesis”and“steroid metabolic process”indicated the presence of endoplasmic reticulum stress,which contributes to the development of steatohepatitis.DNA repair pathways and associated terms indicated hepatocyte death.The presence of increased dendritic cells,along with expanded activated dendritic cells in nonalcoholic steatohepatitis(NASH),indicated that the 3 mo socs8^(-/-)model may represent the late stage of MAFLD.In the gut,imaging results from larvae and single-cell sequencing of intestinal mucosa cells from 3 mo socs8^(-/-)fish revealed a consistent accumulation of lymphocytes,with the number of innate immune cells higher in larvae but lower in adult fish,compared to wide type fish.In the 1 mo socs8^(-/-)fish,enhanced mucin secretion,suggested by a greater number of goblet cells and upregulated“SNARE interactions in vesicular transport”,was in line with“mannose type O-glycan biosynthesis”.The same group had upregulated IL-13 and its receptors,aligning with the GO term“wound healing”and the idea that innate lym-phocytes producing IL-13 may function as regulators of immunity and tissue remodeling.The enriched pathways“arachidonic acid metabolism”and“intestinal immune network for IgA production”at 1 month,indicating enhanced proliferation of intestinal epithelial cells and improved host‒microbe interactions,respectively.Comparing 1 mo socs8^(-/-)with wide type fish,higher levels of Prevotella indicate enhanced glucose metabolism due to their role in polysaccharide breakdown.Meanwhile,lower levels of Veillonella indicate oxidative stress,potentially associated with a reduced lumen at one month.Intestinal bacterial overgrowth in 3 mo socs8^(-/-)fish(compared to 1 mo socs8^(-/-))contributes to the exacerbation of liver dysfunction.Compared 3 mo socs8^(-/-)with wide type fish,despite an increase in T/NK cells,elevated fatty acid metabolism may impair the function of intestinal lymphocytes.Therefore,the symptoms observed in the gut‒liver axis suggest that socs8^(-/-)zebrafish may serve as a model for MAFLD.
基金funded by the Supporting Project for the Development of Industrial Science and Technology,Jiangxi Innovation and Incubation Center of Industrial Technologies,Chinese Academy of Sciences(ZKYJXZXCY-202210)as well as the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA24010206).
文摘This study examined the effects of Picria fel-terrae Lour.(PFL)on soybean meal-induced enteritis in zebrafish(Danio rerio)and evaluated whether its increased resistance to Aeromonas hydrophila using both in vivo and in vitro approaches.Compared to plain soybean meal,the addition of 0.05%,0.1%,or 0.2%PFL to zebrafish larval diet reduced intestinal neutrophil(0.1%and 0.2%PFL),macrophage(0.05%and 0.1%PFL),and T cell aggregation(0.01%,0.05%and 0.1%PFL).Following a six-week feeding trial with 0.1%PFL,the hindgut of 3-month-old zebrafish showed a higher length/width ratio of intestinal ridges and the regulation of macrophages and CD4+cells compared to fish fed soybean meal.Immune cell infiltration analysis also demonstrated immune modulatory effects of PFL,such as decreased activated memory CD4 T cells and increased M2 macrophages,and we observed a reduction in nuclear density in the liver,accompanied by an increase in PAS-stained glycogen and p-STAT3 signals.Enrichment analyses revealed that PFL may mitigate intestinal inflammation by upregulating genes involved in DNA replication and downregulating inflammatory processes,such as leukocyte chemotaxis,lysosomes,and phagosomes.The enriched hepatic terms and pathways indicated that PFL facilitated the metabolism of fatty acids and monocarboxylic acids while inhibiting the alteration of tight junctions and the generation of reactive oxygen species by NADPH oxidases.Hexanorcucurbitacin F and its target genes were revealed by network pharmacological analysis and molecular docking,and its target proteins Src and Ptk2ab were related to macrophage function.PFL increased Bifidobacterium longum and species of Faecalitalea,which may contribute to gut homeostasis.During the Aeromonas hydrophila challenge,PFL resulted in a higher survival rate and reduced swelling of the intrabronchial lymphoid tissue compared to soybean meal.PFL water extract also inhibited the growth of A.hydrophila in vitro.In conclusion,PFL dietary inclusion restored intestinal im-mune homeostasis by modulating the function and aggregation of immune cells,stimulating cell regeneration,rebalancing gut flora,and controlling hepatic metaflammation.These findings demonstrate that PFL enhances immune responses across the gut-liver axis and improves mucosal antibacterial defenses in fish,and is a potential functional feed additive.
