The aim of the present study was to explore the effects of annexin A1(ANXA1) mimetic peptide AC2-26 on sepsis-induced cardiomyocyte apoptosis in vivo and in vitro and the underlying mechanisms.In the in vivo study,a r...The aim of the present study was to explore the effects of annexin A1(ANXA1) mimetic peptide AC2-26 on sepsis-induced cardiomyocyte apoptosis in vivo and in vitro and the underlying mechanisms.In the in vivo study,a rat septic model was established by the cecal ligation and puncture (CLP).The rats were divided into control group,sepsis group and AC2-26 group.The rats in the AC2-26 group were intraperitoneally injected with AC2-26(1mg/kg)2h before CLP,and those in the control group and sepsis group were injected with the same volume of normal saline.The myocardial tissue was examined by hematoxylin and eosin (HE)staining and transmission electron microscopy (TEM).Furthermore,myocardial apoptosis was measured by terminal dUTP nick end-labeling (TUNEL)assay.In the in vitro study,H9C2cells were cultured and divided into three groups:control group,in which cells were only given the basic culture medium;LPS group,in which cells were treated with 10μg/mL LPS;AC2-26 group,in which cells were treated with 0.5μmol/L AC2-262h before 10μg/mL LPS was given.The apoptosis of H9C2 cells was detected by flow cytometry.The levels of lipoxin A4 receptor (LXA4),phosphoinositide3-kinase (PI3K)and protein kinase B (PKB or AKT)protein were measured by Western blotting, the activity of NF-KB and the level of TNF-α by ELISA and the activities of caspase-3/8by using the caspase activity kits.The in vivo study showed that the myocardial pathological damage and myocardial ultrastructural damage were significantly alleviated and the myocardial apoptosis significantly decreased in the AC2-26 group as compared with the sepsis group (P<0.05 for all). The in vivo study revealed that the apoptosis of H9C2 cells was profoundly ameliorated in the AC2-26 group relative to the sepsis group (P<0.05).The protein expression levels of LXA4 were significantly up-regulated,and those of PI3K and AKT prominently down-regulated in the AC2-26 group when compared with those in the LPS group (P<0.05 for all).The activity of NF-κB was greatly inhibited and the level of TNF-α markedly decreased in the AC2-26 group as compared with those in the LPS group (P<0.05 for all).AC2-26 treatment also significantly suppressed the activities of caspase-3/8 in H9C2 cells.In conclusion,these findings suggest that AC2-26 may alleviate the sepsis-induced cardiomyocyte apoptosis in vivo and in vivo through the LXA4/PI3K/ AKT signaling pathway.展开更多
Rheumatoid arthritis(RA)is a common autoimmune disease characterized by joint inflammation and immune dysfunction.Although various therapeutic approaches have been utilized for the treatment of RA in clinical applicat...Rheumatoid arthritis(RA)is a common autoimmune disease characterized by joint inflammation and immune dysfunction.Although various therapeutic approaches have been utilized for the treatment of RA in clinical applications,the low responsiveness of RA patients and undesired systemic toxicity are still unresolved problems.Targeting the resolution pathway of inflammation with pro-resolving mediators would evoke the protective actions of patient for combating the inflammation.Ac2–26,a 25-amino acid peptide derived from Annexin A(a pro-resolving mediator),has shown good efficacy in the treatment of inflammatory disorders.However,the low bioavailability of Ac2–26 peptides hinders their efficacy in vivo.In this paper,we formed PEGylated lipid nanoparticles(LDNPs)by the co-assembly of l-ascorbyl palmitate(L-AP)and N-(carbonyl methoxypolyethylene glycol-2000)-1,2-distearoyl-sn–glycero-3-phosphoethanolamine(DSPE-PEG 2 k)to encapsulate and deliver Ac2–26 peptides to the arthritic rats.They showed good stability and biocompatibility.After being intravenously administrated,Ac2–26 peptide-loaded PEGylated lipid nanoparticles(ADNPs)showed the prolonged in vivo circulation time and enhanced accumulation in inflamed sites.In vivo therapeutic evaluations revealed that ADNPs could attenuate synovial inflammation and improve joint pathology.Therefore,the pro-resolving therapeutic strategy using ADNPs is effective in RA treatment.展开更多
Tomato yellow leaf curl China virus Y10 isolate (TYLCCNV-Y10) alone could systemically infect host plants such as Nicotiana benthamiana without symptoms. In con- trast, Tobacco curly shoot virus Y35 isolate (TbCSV-Y35...Tomato yellow leaf curl China virus Y10 isolate (TYLCCNV-Y10) alone could systemically infect host plants such as Nicotiana benthamiana without symptoms. In con- trast, Tobacco curly shoot virus Y35 isolate (TbCSV-Y35) alone induces leaf curl symptoms in N. benthamiana. When inoculated into transgenic N. benthamiana plants expressing GFP gene (line 16c), TYLCCNV-Y10 neither reverses the established GFP silencing nor blocks the onset of GFP si- lencing. In contrast, TbCSV-Y35 can partially reverse the established GFP silencing and block the onset of GFP silenc- ing in new leaves. In the patch co-infiltration assays, the AC2 and AC4 proteins of TYLCCNV-Y10 and TbCSV-Y35 could suppress local GFP silencing and delay systemic GFP silenc- ing, suggesting that they are suppressors of RNA silencing. Comparison of the accumulation levels of GFP mRNA in the co-infiltration patches showed that Y10 AC2 and Y35 AC2 proteins had similar efficiency for suppression of RNA si- lencing. However, Y35 AC4 protein functioned as a stronger suppressor of RNA silencing than Y10 AC4 protein. There- fore, the pathogenicity difference between TbCSV-Y35 and TYLCCNV-Y10 may be related to the functional difference in their AC4 proteins.展开更多
基金the Hubei Provincial Natural Science Foundation(No.2018CFC847)the Hubei Cancer Hospital Foundation(No.2015C11).
文摘The aim of the present study was to explore the effects of annexin A1(ANXA1) mimetic peptide AC2-26 on sepsis-induced cardiomyocyte apoptosis in vivo and in vitro and the underlying mechanisms.In the in vivo study,a rat septic model was established by the cecal ligation and puncture (CLP).The rats were divided into control group,sepsis group and AC2-26 group.The rats in the AC2-26 group were intraperitoneally injected with AC2-26(1mg/kg)2h before CLP,and those in the control group and sepsis group were injected with the same volume of normal saline.The myocardial tissue was examined by hematoxylin and eosin (HE)staining and transmission electron microscopy (TEM).Furthermore,myocardial apoptosis was measured by terminal dUTP nick end-labeling (TUNEL)assay.In the in vitro study,H9C2cells were cultured and divided into three groups:control group,in which cells were only given the basic culture medium;LPS group,in which cells were treated with 10μg/mL LPS;AC2-26 group,in which cells were treated with 0.5μmol/L AC2-262h before 10μg/mL LPS was given.The apoptosis of H9C2 cells was detected by flow cytometry.The levels of lipoxin A4 receptor (LXA4),phosphoinositide3-kinase (PI3K)and protein kinase B (PKB or AKT)protein were measured by Western blotting, the activity of NF-KB and the level of TNF-α by ELISA and the activities of caspase-3/8by using the caspase activity kits.The in vivo study showed that the myocardial pathological damage and myocardial ultrastructural damage were significantly alleviated and the myocardial apoptosis significantly decreased in the AC2-26 group as compared with the sepsis group (P<0.05 for all). The in vivo study revealed that the apoptosis of H9C2 cells was profoundly ameliorated in the AC2-26 group relative to the sepsis group (P<0.05).The protein expression levels of LXA4 were significantly up-regulated,and those of PI3K and AKT prominently down-regulated in the AC2-26 group when compared with those in the LPS group (P<0.05 for all).The activity of NF-κB was greatly inhibited and the level of TNF-α markedly decreased in the AC2-26 group as compared with those in the LPS group (P<0.05 for all).AC2-26 treatment also significantly suppressed the activities of caspase-3/8 in H9C2 cells.In conclusion,these findings suggest that AC2-26 may alleviate the sepsis-induced cardiomyocyte apoptosis in vivo and in vivo through the LXA4/PI3K/ AKT signaling pathway.
基金supported by the National Natural Science Foundation of China(No.82003661)。
文摘Rheumatoid arthritis(RA)is a common autoimmune disease characterized by joint inflammation and immune dysfunction.Although various therapeutic approaches have been utilized for the treatment of RA in clinical applications,the low responsiveness of RA patients and undesired systemic toxicity are still unresolved problems.Targeting the resolution pathway of inflammation with pro-resolving mediators would evoke the protective actions of patient for combating the inflammation.Ac2–26,a 25-amino acid peptide derived from Annexin A(a pro-resolving mediator),has shown good efficacy in the treatment of inflammatory disorders.However,the low bioavailability of Ac2–26 peptides hinders their efficacy in vivo.In this paper,we formed PEGylated lipid nanoparticles(LDNPs)by the co-assembly of l-ascorbyl palmitate(L-AP)and N-(carbonyl methoxypolyethylene glycol-2000)-1,2-distearoyl-sn–glycero-3-phosphoethanolamine(DSPE-PEG 2 k)to encapsulate and deliver Ac2–26 peptides to the arthritic rats.They showed good stability and biocompatibility.After being intravenously administrated,Ac2–26 peptide-loaded PEGylated lipid nanoparticles(ADNPs)showed the prolonged in vivo circulation time and enhanced accumulation in inflamed sites.In vivo therapeutic evaluations revealed that ADNPs could attenuate synovial inflammation and improve joint pathology.Therefore,the pro-resolving therapeutic strategy using ADNPs is effective in RA treatment.
文摘Tomato yellow leaf curl China virus Y10 isolate (TYLCCNV-Y10) alone could systemically infect host plants such as Nicotiana benthamiana without symptoms. In con- trast, Tobacco curly shoot virus Y35 isolate (TbCSV-Y35) alone induces leaf curl symptoms in N. benthamiana. When inoculated into transgenic N. benthamiana plants expressing GFP gene (line 16c), TYLCCNV-Y10 neither reverses the established GFP silencing nor blocks the onset of GFP si- lencing. In contrast, TbCSV-Y35 can partially reverse the established GFP silencing and block the onset of GFP silenc- ing in new leaves. In the patch co-infiltration assays, the AC2 and AC4 proteins of TYLCCNV-Y10 and TbCSV-Y35 could suppress local GFP silencing and delay systemic GFP silenc- ing, suggesting that they are suppressors of RNA silencing. Comparison of the accumulation levels of GFP mRNA in the co-infiltration patches showed that Y10 AC2 and Y35 AC2 proteins had similar efficiency for suppression of RNA si- lencing. However, Y35 AC4 protein functioned as a stronger suppressor of RNA silencing than Y10 AC4 protein. There- fore, the pathogenicity difference between TbCSV-Y35 and TYLCCNV-Y10 may be related to the functional difference in their AC4 proteins.
