Background Necrotic enteritis(NE)in broiler chickens leads to significant economic losses in poultry production.This study examined the inhibitory effects of usnic acid and tannic acid on coccidia,sporozoite,and Clost...Background Necrotic enteritis(NE)in broiler chickens leads to significant economic losses in poultry production.This study examined the inhibitory effects of usnic acid and tannic acid on coccidia,sporozoite,and Clostridium perfringens and assessed their influence on growth performance and intestinal health in NE-challenged broilers through in vitro and in vivo experiments.Methods The in vitro experiment included 5 treatment groups:the negative control(NC),2μmol/L diclazuril(DZ),30μmol/L usnic acid(UA),90μmol/L tannic acid(TA),and 15μmol/L usnic acid^(+)45μmol/L tannic acid(UTA)groups.The in vivo experiment involved 320 broilers divided into four groups:PC(NE-challenged),SA(500 mg/kg salinomycin premix^(+)NE-challenged),UA(300 mg/kg usnic acid^(+)NE-challenged),and UTA(300 mg/kg usnic acid^(+)500 mg/kg tannic acid^(+)NE-challenged)groups.Results In the in vitro study,the UA,TA,and UTA treatments significantly increased apoptosis in coccidian oocysts and sporozoites,lowered the mitochondrial membrane potential(P<0.05),and disrupted the oocyst structure compared with those in the NC group.UA and TA had inhibitory effects on C.perfringens,with the strongest inhibition observed in the UTA group.The in vivo results demonstrated that the SA group presented significantly improved growth performance on d 13,21,and 28(P<0.05),whereas the UA and UTA groups presented improvements on d 13 and 21(P<0.05).The SA,UA,and UTA treatments reduced the intestinal lesion scores by d 28 and the fecal coccidian oocyst counts from d 19 to 21(P<0.05).Compared with the PC group,the UA and UTA groups presented lower intestinal sIgA levels and CD8^(+)cell percentages(P<0.05),with a trend toward a reduced CD3^(+)cell percentage(P=0.069).The SA,UA,and UTA treatments significantly reduced the serum diamine oxidase activity,crypt depth,and plateletderived growth factor levels in the intestinal mucosa while increasing the villus height to crypt depth ratio and number of goblet cells(P<0.05).The UTA treatment also significantly increased the acetate and butyrate concentrations in the cecum(P<0.05).With respect to the gut microbiota,significant changes inβdiversity in the ileum and cecum were observed in the SA,UA,and UTA groups,indicating that the microbial community compositions differed among the groups.Romboutsia dominated the SA group,Bacillales dominated the UA group,and Lactobacillales and Lachnospirales dominated the UTA group in the ileal microbiota.In the cecal microbiota,Lactobacillus,Butyricicoccus,and Blautia abundances were significantly elevated in the UTA group(P<0.05).Conclusion Usnic acid and tannic acid induce apoptosis in coccidia and sporozoites by lowering the mitochondrial membrane potential.Both usnic acid alone and in combination with tannic acid alleviate NE-induced adverse effects in broilers by modulating intestinal immunity,altering the microbial composition,and improving intestinal barrier function.Compared with usnic acid alone,the combination of usnic acid and tannic acid had superior effects,providing a promising basis for the development of effective feed additive combinations.展开更多
The anti-inflammatory effect and mechanism of Usnic acid （UA） were explored on lipopoly-saccharide （LPS）-stimulated RAW264.7 cell line.The effects of UA on pro-inflammatory cytokines including tumor necrosis facto...The anti-inflammatory effect and mechanism of Usnic acid （UA） were explored on lipopoly-saccharide （LPS）-stimulated RAW264.7 cell line.The effects of UA on pro-inflammatory cytokines including tumor necrosis factor-alfa （TNF-α）,interleukin-6 （IL-6） and interleukin-1 beta （IL-1β）,pro-inflammatory mediators such as nitric oxide （NO）,inducible nitric oxide synthase （iNOS） and cyclooxygenase-2 （COX-2） were studied by sandwich ELISA,real-time PCR and western blot analyses.Similarly,the effect of UA on anti-inflammatory cytokine interleukin-10 （IL-10） and anti-inflammatory mediator heme oxygenase-1 （HO-1） were also studied following the same methods.Furthermore,nuclear factor-κB （NF-κB） was assayed by immunocytochemistry.The results showed that UA has anti-inflammatory effect by down-regulatinng iNOS,COX-2,IL-1β,IL-6 and TNF-α,COX-2 gene expression through the suppression of NF-κB activation and increasing anti-inflammatory cytokine IL-10 and anti-inflammatory mediator HO-1 production.展开更多
Objective To investigate the molecular mechanisms that are responsible for anti-inflammatory effect of usnic acid (UA), the effects of UA from usnea longissm on tumor necrosis factor-α(TNF-α) and nitric oxide (NO) p...Objective To investigate the molecular mechanisms that are responsible for anti-inflammatory effect of usnic acid (UA), the effects of UA from usnea longissm on tumor necrosis factor-α(TNF-α) and nitric oxide (NO) production in peritoneal macrophages has been examined. Methods The different concentrations of UA were added to peritoneal macrophages. The TNF-α and NO production in peritoneal macrophages were examined with mouse TNF-α ELISA kit and NO content by measuring the amount of nitrite (NO-_2μmol/L) formed in the medium using Griess reaction. The activity of inducible nitric oxide synthase (i-NOS) was determined using i-NOS detection kit and the TNF-α mRNA expression was tested by reverse transcriptase polymerase chain reaction (RT-PCR). Results UA decreased the TNF-α and NO level in LPS-stimulated peritoneal macrophages in dose-dependent manner, the IC_ 50 values were 12.8μmol/L and 5.7μmol/L respectively. RT-PCR analysis indicated that UA could inhibit TNF-α mRNA expression; the activity analysis of i-NOS indicated that UA could inhibit the activity of i-NOS. Conclusion UA could inhibit the TNF-α and NO production in peritoneal macrophages, it may be associated with the anti-inflammatory activity of UA.展开更多
The normal fine structures of toxoplasma gondii tachyzoite and the erfect of usnic acid on the ultrastructure of the tachyzoites were observed by transmission electron microscope (TEM).The studies indicate that the pa...The normal fine structures of toxoplasma gondii tachyzoite and the erfect of usnic acid on the ultrastructure of the tachyzoites were observed by transmission electron microscope (TEM).The studies indicate that the pathological changes or the ultrastructure of the parasite took place under the effect of the 10 mg/L usnic acid for 30 min.These changes can be illustrated as folio'vs; 1)The posterior portion of the rhopties were destroyed.2)The membrane of the daughter cell fractured. 3)The membranate organellae or the organisms were demaged.展开更多
This paper reports the ultrastructural changes of trichomonas vaginaiis (T. vag. ) under the action of s-(-)usnic acid sodium in vitro. These changes can be shown by the following results:At first, the degranulation o...This paper reports the ultrastructural changes of trichomonas vaginaiis (T. vag. ) under the action of s-(-)usnic acid sodium in vitro. These changes can be shown by the following results:At first, the degranulation of the rough endoplasmic reticulum (RER) took place, the polyribosomes disaggregated. And then, the Golgi complex and the endoplasmic reticulum dilated. The cytoplasmic matrix presented and inhomogeneous apperance. Finally, the biomembrane loosed and fractured.The cell nuclei presented the karyorrhexis.展开更多
基金supported by China Agriculture Research System Program(Project No.CARS-41-G04)。
文摘Background Necrotic enteritis(NE)in broiler chickens leads to significant economic losses in poultry production.This study examined the inhibitory effects of usnic acid and tannic acid on coccidia,sporozoite,and Clostridium perfringens and assessed their influence on growth performance and intestinal health in NE-challenged broilers through in vitro and in vivo experiments.Methods The in vitro experiment included 5 treatment groups:the negative control(NC),2μmol/L diclazuril(DZ),30μmol/L usnic acid(UA),90μmol/L tannic acid(TA),and 15μmol/L usnic acid^(+)45μmol/L tannic acid(UTA)groups.The in vivo experiment involved 320 broilers divided into four groups:PC(NE-challenged),SA(500 mg/kg salinomycin premix^(+)NE-challenged),UA(300 mg/kg usnic acid^(+)NE-challenged),and UTA(300 mg/kg usnic acid^(+)500 mg/kg tannic acid^(+)NE-challenged)groups.Results In the in vitro study,the UA,TA,and UTA treatments significantly increased apoptosis in coccidian oocysts and sporozoites,lowered the mitochondrial membrane potential(P<0.05),and disrupted the oocyst structure compared with those in the NC group.UA and TA had inhibitory effects on C.perfringens,with the strongest inhibition observed in the UTA group.The in vivo results demonstrated that the SA group presented significantly improved growth performance on d 13,21,and 28(P<0.05),whereas the UA and UTA groups presented improvements on d 13 and 21(P<0.05).The SA,UA,and UTA treatments reduced the intestinal lesion scores by d 28 and the fecal coccidian oocyst counts from d 19 to 21(P<0.05).Compared with the PC group,the UA and UTA groups presented lower intestinal sIgA levels and CD8^(+)cell percentages(P<0.05),with a trend toward a reduced CD3^(+)cell percentage(P=0.069).The SA,UA,and UTA treatments significantly reduced the serum diamine oxidase activity,crypt depth,and plateletderived growth factor levels in the intestinal mucosa while increasing the villus height to crypt depth ratio and number of goblet cells(P<0.05).The UTA treatment also significantly increased the acetate and butyrate concentrations in the cecum(P<0.05).With respect to the gut microbiota,significant changes inβdiversity in the ileum and cecum were observed in the SA,UA,and UTA groups,indicating that the microbial community compositions differed among the groups.Romboutsia dominated the SA group,Bacillales dominated the UA group,and Lactobacillales and Lachnospirales dominated the UTA group in the ileal microbiota.In the cecal microbiota,Lactobacillus,Butyricicoccus,and Blautia abundances were significantly elevated in the UTA group(P<0.05).Conclusion Usnic acid and tannic acid induce apoptosis in coccidia and sporozoites by lowering the mitochondrial membrane potential.Both usnic acid alone and in combination with tannic acid alleviate NE-induced adverse effects in broilers by modulating intestinal immunity,altering the microbial composition,and improving intestinal barrier function.Compared with usnic acid alone,the combination of usnic acid and tannic acid had superior effects,providing a promising basis for the development of effective feed additive combinations.
文摘The anti-inflammatory effect and mechanism of Usnic acid （UA） were explored on lipopoly-saccharide （LPS）-stimulated RAW264.7 cell line.The effects of UA on pro-inflammatory cytokines including tumor necrosis factor-alfa （TNF-α）,interleukin-6 （IL-6） and interleukin-1 beta （IL-1β）,pro-inflammatory mediators such as nitric oxide （NO）,inducible nitric oxide synthase （iNOS） and cyclooxygenase-2 （COX-2） were studied by sandwich ELISA,real-time PCR and western blot analyses.Similarly,the effect of UA on anti-inflammatory cytokine interleukin-10 （IL-10） and anti-inflammatory mediator heme oxygenase-1 （HO-1） were also studied following the same methods.Furthermore,nuclear factor-κB （NF-κB） was assayed by immunocytochemistry.The results showed that UA has anti-inflammatory effect by down-regulatinng iNOS,COX-2,IL-1β,IL-6 and TNF-α,COX-2 gene expression through the suppression of NF-κB activation and increasing anti-inflammatory cytokine IL-10 and anti-inflammatory mediator HO-1 production.
基金This work was supported by the Natural Science Foundation of Shaanxi Province ( No.2004G240) .
文摘Objective To investigate the molecular mechanisms that are responsible for anti-inflammatory effect of usnic acid (UA), the effects of UA from usnea longissm on tumor necrosis factor-α(TNF-α) and nitric oxide (NO) production in peritoneal macrophages has been examined. Methods The different concentrations of UA were added to peritoneal macrophages. The TNF-α and NO production in peritoneal macrophages were examined with mouse TNF-α ELISA kit and NO content by measuring the amount of nitrite (NO-_2μmol/L) formed in the medium using Griess reaction. The activity of inducible nitric oxide synthase (i-NOS) was determined using i-NOS detection kit and the TNF-α mRNA expression was tested by reverse transcriptase polymerase chain reaction (RT-PCR). Results UA decreased the TNF-α and NO level in LPS-stimulated peritoneal macrophages in dose-dependent manner, the IC_ 50 values were 12.8μmol/L and 5.7μmol/L respectively. RT-PCR analysis indicated that UA could inhibit TNF-α mRNA expression; the activity analysis of i-NOS indicated that UA could inhibit the activity of i-NOS. Conclusion UA could inhibit the TNF-α and NO production in peritoneal macrophages, it may be associated with the anti-inflammatory activity of UA.
文摘The normal fine structures of toxoplasma gondii tachyzoite and the erfect of usnic acid on the ultrastructure of the tachyzoites were observed by transmission electron microscope (TEM).The studies indicate that the pathological changes or the ultrastructure of the parasite took place under the effect of the 10 mg/L usnic acid for 30 min.These changes can be illustrated as folio'vs; 1)The posterior portion of the rhopties were destroyed.2)The membrane of the daughter cell fractured. 3)The membranate organellae or the organisms were demaged.
文摘This paper reports the ultrastructural changes of trichomonas vaginaiis (T. vag. ) under the action of s-(-)usnic acid sodium in vitro. These changes can be shown by the following results:At first, the degranulation of the rough endoplasmic reticulum (RER) took place, the polyribosomes disaggregated. And then, the Golgi complex and the endoplasmic reticulum dilated. The cytoplasmic matrix presented and inhomogeneous apperance. Finally, the biomembrane loosed and fractured.The cell nuclei presented the karyorrhexis.
