Objective:The effect of casein glycomacropeptide(CGMP)on the expression of NF-B subunit p65 in human colorectal cancer HT-29 cells induced by lipopolysaccharide(LPS)was investigated to explore the therapeutic efficacy...Objective:The effect of casein glycomacropeptide(CGMP)on the expression of NF-B subunit p65 in human colorectal cancer HT-29 cells induced by lipopolysaccharide(LPS)was investigated to explore the therapeutic efficacy of CGMP for human colorectal cancer.Methods:HT-29 cells cultured in 96-well plates were stimulated with LPS for 30 min at the concentrations of 0,0.001,0.01,0.1,1 and 10g/mL.After stimulation,the expression of p65 in HT-29 cells was evaluated by immunofluorescence method.Similarly,HT-29 cells were incubated with 0,0.001,0.01,0.1,1,10 and 100g/mL CGMP for 24 h at the optimal LPS concentration.After treatment with CGMP under the incubation with optimal LPS concentration for 30 min,the protein expression of p65 was analyzed by Western blotting.The optimum culture time was evaluated by incubating HT-29 cells with the optimal CGMP concentration for 6,12,24,48 and 72 h.Results:NF-B-p65 revealed that the highest protein expression was achieved with 1g/mL LPS treatment.Meanwhile,CGMP could inhibit the protein expression of NF-B-p65 in LPS-stimulated HT-29 cells,and the optimal inhibitory effect was observed at a CGMP concentration of 0.01g/mL with 48 h incubation.Conclusion:CGMP can regulate NF-B signaling pathway through inhibiting the expression of its subunit p65,which is beneficial for the further improvement of human colorectal cancer treatment.展开更多
Powdery mildew(PM)is considered a major cause of yield losses and reduced quality in cucumber worldwide,but the molecular basis of PM resistance remains poorly understood.A segment substitution line,namely,SSL508-28,w...Powdery mildew(PM)is considered a major cause of yield losses and reduced quality in cucumber worldwide,but the molecular basis of PM resistance remains poorly understood.A segment substitution line,namely,SSL508-28,was developed with dominant PM resistance in the genetic background of PM-susceptible cucumber inbred line D8.The substituted segment contains 860 genes.An iTRAQ-based comparative proteomic technology was used to map the proteomes of PM-inoculated and untreated(control)D8 and SSL508-28.The number of differentially regulated proteins(DRPs)in SSL508-28 was almost three times higher than that in D8.Fourteen DRPs were located in the substituted segment interval.Comparative gene expression analysis revealed that nodulin-related protein 1(NRP1)may be a good candidate for PM resistance.Gene Ontology enrichment analysis showed that DRPs functioning in tetrapyrrole biosynthetic process,sulfur metabolic process and cell redox homeostasis were specifically enriched in the resistant line SSL508-28.DRPs categorized in the KEGG term photosynthesis increased in both lines upon PM infection,suggesting that the strategies used by cucumber may be different from those used by other crops to react to PM attacks at the initial stage.The measurement of hydrogen peroxide and superoxide anion production and net photosynthetic rate were consistent with the changes in protein abundance,suggesting that the proteomic results were reliable.There was a poor correlation between DRPs measured by iTRAQ and the corresponding gene expression changes measured by RNA-seq with the same experimental design.Taken together,these findings improve the understanding of the molecular mechanisms underlying the response of cucumber to PM infection.展开更多
Diarrhea is considered to be associated with microbial dysbiosis caused by infection of pathogens but poorly understood.We herein characterized the colonic microbiota of diarrheal early-weaning piglets infected with p...Diarrhea is considered to be associated with microbial dysbiosis caused by infection of pathogens but poorly understood.We herein characterized the colonic microbiota of diarrheal early-weaning piglets infected with porcine circovirus type 2(PCV2)and Campylobacter.Campylobacter infection significantly decreased species richness and Shannon diversity index of colonic microbiota together with a significant increase in the proportion of Campylobacter and Enterobacteriaceae,whereas no significant difference on the above indexes was observed in piglets infected with PCV2 compared with healthy piglets,PCV2 and Campylobacter infection could disturb the homeostasis of colonic microbiota through deterioration of ecological network within microbial community,and specially Campylobacter performed as a module hub in ecological networks.The microbial dysbiosis caused metabolic dysfunction and led to a remarkable reduction in production of short chain fatty acids,following by a higher pH level in colon cavity.Campylobacter infection disturbed the function of colonic tract barrier observed in terms of significant lower relative expression of claudin-1,occluding,and zonula occludens protein-1 genes,and PCV2 infection induced intestinal inflammation together with a higher permeability of colon.Generally,these results suggested that PCV2 and Campylobacter infection could induce microbial dysbiosis and metabolic dysfunction,and cause intestinal disorder,all of which finally were associated to contribute to the diarrhea of early-weaning piglets.展开更多
文摘Objective:The effect of casein glycomacropeptide(CGMP)on the expression of NF-B subunit p65 in human colorectal cancer HT-29 cells induced by lipopolysaccharide(LPS)was investigated to explore the therapeutic efficacy of CGMP for human colorectal cancer.Methods:HT-29 cells cultured in 96-well plates were stimulated with LPS for 30 min at the concentrations of 0,0.001,0.01,0.1,1 and 10g/mL.After stimulation,the expression of p65 in HT-29 cells was evaluated by immunofluorescence method.Similarly,HT-29 cells were incubated with 0,0.001,0.01,0.1,1,10 and 100g/mL CGMP for 24 h at the optimal LPS concentration.After treatment with CGMP under the incubation with optimal LPS concentration for 30 min,the protein expression of p65 was analyzed by Western blotting.The optimum culture time was evaluated by incubating HT-29 cells with the optimal CGMP concentration for 6,12,24,48 and 72 h.Results:NF-B-p65 revealed that the highest protein expression was achieved with 1g/mL LPS treatment.Meanwhile,CGMP could inhibit the protein expression of NF-B-p65 in LPS-stimulated HT-29 cells,and the optimal inhibitory effect was observed at a CGMP concentration of 0.01g/mL with 48 h incubation.Conclusion:CGMP can regulate NF-B signaling pathway through inhibiting the expression of its subunit p65,which is beneficial for the further improvement of human colorectal cancer treatment.
基金supported by the National Natural Science Foundation of China(grant nos.31672176 and 31171978)the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(18KJB210014).
文摘Powdery mildew(PM)is considered a major cause of yield losses and reduced quality in cucumber worldwide,but the molecular basis of PM resistance remains poorly understood.A segment substitution line,namely,SSL508-28,was developed with dominant PM resistance in the genetic background of PM-susceptible cucumber inbred line D8.The substituted segment contains 860 genes.An iTRAQ-based comparative proteomic technology was used to map the proteomes of PM-inoculated and untreated(control)D8 and SSL508-28.The number of differentially regulated proteins(DRPs)in SSL508-28 was almost three times higher than that in D8.Fourteen DRPs were located in the substituted segment interval.Comparative gene expression analysis revealed that nodulin-related protein 1(NRP1)may be a good candidate for PM resistance.Gene Ontology enrichment analysis showed that DRPs functioning in tetrapyrrole biosynthetic process,sulfur metabolic process and cell redox homeostasis were specifically enriched in the resistant line SSL508-28.DRPs categorized in the KEGG term photosynthesis increased in both lines upon PM infection,suggesting that the strategies used by cucumber may be different from those used by other crops to react to PM attacks at the initial stage.The measurement of hydrogen peroxide and superoxide anion production and net photosynthetic rate were consistent with the changes in protein abundance,suggesting that the proteomic results were reliable.There was a poor correlation between DRPs measured by iTRAQ and the corresponding gene expression changes measured by RNA-seq with the same experimental design.Taken together,these findings improve the understanding of the molecular mechanisms underlying the response of cucumber to PM infection.
基金supported by National Key R&D Program of China(2016YFD0501201)China Scholarship Council(201806825084)
文摘Diarrhea is considered to be associated with microbial dysbiosis caused by infection of pathogens but poorly understood.We herein characterized the colonic microbiota of diarrheal early-weaning piglets infected with porcine circovirus type 2(PCV2)and Campylobacter.Campylobacter infection significantly decreased species richness and Shannon diversity index of colonic microbiota together with a significant increase in the proportion of Campylobacter and Enterobacteriaceae,whereas no significant difference on the above indexes was observed in piglets infected with PCV2 compared with healthy piglets,PCV2 and Campylobacter infection could disturb the homeostasis of colonic microbiota through deterioration of ecological network within microbial community,and specially Campylobacter performed as a module hub in ecological networks.The microbial dysbiosis caused metabolic dysfunction and led to a remarkable reduction in production of short chain fatty acids,following by a higher pH level in colon cavity.Campylobacter infection disturbed the function of colonic tract barrier observed in terms of significant lower relative expression of claudin-1,occluding,and zonula occludens protein-1 genes,and PCV2 infection induced intestinal inflammation together with a higher permeability of colon.Generally,these results suggested that PCV2 and Campylobacter infection could induce microbial dysbiosis and metabolic dysfunction,and cause intestinal disorder,all of which finally were associated to contribute to the diarrhea of early-weaning piglets.
