Background Rheumatoid arthritis （RA） is an autoimmune disease characterized by chronic inflammation at the synovial membrane. Although great progress has been made recently in exploring the etiology and pathogenesis...Background Rheumatoid arthritis （RA） is an autoimmune disease characterized by chronic inflammation at the synovial membrane. Although great progress has been made recently in exploring the etiology and pathogenesis of RA, its molecular pathological mechanism remains to be further defined and it is still a great challenge in determining the diagnosis and in choosing the appropriate therapy in early patients. This study was performed to screen candidate RA-associated serum proteins by comparative proteomics to provide research clues to early diagnosis and treatment of RA. Methods Sera isolated from 6 RA patients and 6 healthy volunteers were pooled respectively and high-abundance proteins were depleted by Plasma 7 Multiple Affinity Removal System. The protein expression profiles between the two groups were then compared by two-dimensional gel electrophoresis （2-DE） and the proteins over/under-expressed by more than 3-fold were identified by mass spectrometry analysis. To validate the differential expression levels of the identified proteins between the two groups, ELISA was performed in two of the identified proteins in individual sera from 32 RA patients and 32 volunteers. Results Eight proteins which over/under-expressed in sera of RA patients were identified. Among them, chain A of transthyretin （TTR） was under-expressed, while serum amyloid A protein, apolipoprotein A （ApoA）-IV, ApoA-IV precursor, haptoglobin 2, ceruloplasmin （Cp）, immunoglobulin superfamily 22 and HT016 were over-expressed. ELISA test confirmed that Cp expressed remarkably higher while TTR obviously lower in RA group compared with volunteer group. Conclusion There were 8 identified proteins differentially expressed between RA group and volunteer group, which might be candidate RA-associated proteins and might be promising diagnostic indicators or therapeutic targets for RA.展开更多
Background Ankylosing spondylitis （AS） is a common inflammatory rheumatic disease which lacks satisfactory treatment so far.Sinomenine （SIN） is an alkaloid and has recently been utilized in treating multiple rheum...Background Ankylosing spondylitis （AS） is a common inflammatory rheumatic disease which lacks satisfactory treatment so far.Sinomenine （SIN） is an alkaloid and has recently been utilized in treating multiple rheumatic diseases including AS in China,but its exact mechanism remains to be explored.This study investigated the alteration of proteome in peripheral blood mononuclear cells （PBMCs） from AS patients.Methods Thirty AS patients were enrolled in this study.PBMCs from each AS patient were cultured in medium with or without SIN respectively.Then PBMCs proteins from both groups were separated by two-dimensional electrophoresis （2-DE） and analyzed by mass spectrometry （MS）.Two differentially expressed proteins were then chosen to be verified using Western blotting.Results Seven proteins,including α-synuclein （SNCA）,calmodulin （CALM）,acidic leucine-rich nuclear phosphoprotein 32 family member A （ANP32A）,chloride intracellular channel protein 1 （CLIC1）,guanine nucleotide-binding protein G（I）/G（S）/ G（T） subunit beta-1 （GNB1）,gelsolin （GSN） and histone H2B type 1-M （HISTH2BM）were over-expressed,while coronin1A （CORO1A） was under-expressed in the SIN-treated PBMCs.Further bioinformatics search indicated that the changes of SNCA,ANP32A and CLIC1 pertained to apoptosis,while changes of GSN and CORO1A were associated with both apoptosis and inhibition of immunological function.Subsequently GSN and CORO1A were selected to validate by Western blotting and the results were consistent with those of 2-DE.Conclusion There were 8 differentially expressed proteins in the SIN-treated PBMCs,which might shed some light on the mechanism of SIN in the treatment of AS.展开更多
Background Azithromycin can reduce neutrophil accumulation in neutrophilic pulmonary diseases.However,the precise mechanism behind this action remains unknown.Our experiment assessed whether azithromycin inhibits neut...Background Azithromycin can reduce neutrophil accumulation in neutrophilic pulmonary diseases.However,the precise mechanism behind this action remains unknown.Our experiment assessed whether azithromycin inhibits neutrophil accumulation in the airways by affecting interleukin-17 （IL-17） downstream signals.Methods Mice were pretreated with azithromycin before murine IL-17A （mlL-17） stimulation.After the mlL-17 stimulation,the levels of six neutrophil-mobilizing cytokines were determined by enzyme-linked immunosorbent assay （ELISA） tests in bronchoalveolar lavage （BAL） fluid; IL-6,CXC chemokine ligand-1 （CXCL-1）,CXCL-5,macrophage inflammatory protein-2 （MIP-2）,granulocyte colony-stimulating factor （G-CSF）,and granulocyte macrophage colony-stimulating factor （GM-CSF）.The number of neutrophils in BAL fluid were evaluated by cytospin preparations.Results （1） Azithromycin pretreatment significantly inhibited both the release of three neutrophil-mobilizing cytokines （MIP-2,CXCL-5 and GM-CSF） and the accumulation of neutrophils in airways caused by mlL-17 stimulation.（2） The levels of three neutrophil-mobilizing cytokines （IL-6,MIP-2 and GM-CSF） were positively correlated with the numbers of neutrophil in BAL fluid.Conclusions Azithromycin can inhibit neutrophil accumulation in the airways by affecting IL-17 downstream signals.This finding suggests that macrolide antibiotic application might be useful in prevention of neutrophilic pulmonary diseases characterized by high levels of IL-17.展开更多
基金This study was partly supported by grants from National Natural Science Foundation of China （No. 30325019 and 30471611）, Natural Science Foundation of Guangdong Province （No. 05001703） and Science and Technology Project of Guangdong Province （No. 2008B030301039）.Acknowledgments： We thank the patients and healthy volunteers who participated in this study, and also thank Shanghai Applied Protein Technology Co. Ltd for performing the spectrometry analysis and providing technical support for the 2-DE experiment.
文摘Background Rheumatoid arthritis （RA） is an autoimmune disease characterized by chronic inflammation at the synovial membrane. Although great progress has been made recently in exploring the etiology and pathogenesis of RA, its molecular pathological mechanism remains to be further defined and it is still a great challenge in determining the diagnosis and in choosing the appropriate therapy in early patients. This study was performed to screen candidate RA-associated serum proteins by comparative proteomics to provide research clues to early diagnosis and treatment of RA. Methods Sera isolated from 6 RA patients and 6 healthy volunteers were pooled respectively and high-abundance proteins were depleted by Plasma 7 Multiple Affinity Removal System. The protein expression profiles between the two groups were then compared by two-dimensional gel electrophoresis （2-DE） and the proteins over/under-expressed by more than 3-fold were identified by mass spectrometry analysis. To validate the differential expression levels of the identified proteins between the two groups, ELISA was performed in two of the identified proteins in individual sera from 32 RA patients and 32 volunteers. Results Eight proteins which over/under-expressed in sera of RA patients were identified. Among them, chain A of transthyretin （TTR） was under-expressed, while serum amyloid A protein, apolipoprotein A （ApoA）-IV, ApoA-IV precursor, haptoglobin 2, ceruloplasmin （Cp）, immunoglobulin superfamily 22 and HT016 were over-expressed. ELISA test confirmed that Cp expressed remarkably higher while TTR obviously lower in RA group compared with volunteer group. Conclusion There were 8 identified proteins differentially expressed between RA group and volunteer group, which might be candidate RA-associated proteins and might be promising diagnostic indicators or therapeutic targets for RA.
基金This work was supported by grants from Guangdong Natural Science Foundation （No. 10151008901000010 and No. $2012010011137） and Guangdong Science and Technology Project （No. 2009B080701034）. Conflict of interest： The authors have no conflicts of interest.
文摘Background Ankylosing spondylitis （AS） is a common inflammatory rheumatic disease which lacks satisfactory treatment so far.Sinomenine （SIN） is an alkaloid and has recently been utilized in treating multiple rheumatic diseases including AS in China,but its exact mechanism remains to be explored.This study investigated the alteration of proteome in peripheral blood mononuclear cells （PBMCs） from AS patients.Methods Thirty AS patients were enrolled in this study.PBMCs from each AS patient were cultured in medium with or without SIN respectively.Then PBMCs proteins from both groups were separated by two-dimensional electrophoresis （2-DE） and analyzed by mass spectrometry （MS）.Two differentially expressed proteins were then chosen to be verified using Western blotting.Results Seven proteins,including α-synuclein （SNCA）,calmodulin （CALM）,acidic leucine-rich nuclear phosphoprotein 32 family member A （ANP32A）,chloride intracellular channel protein 1 （CLIC1）,guanine nucleotide-binding protein G（I）/G（S）/ G（T） subunit beta-1 （GNB1）,gelsolin （GSN） and histone H2B type 1-M （HISTH2BM）were over-expressed,while coronin1A （CORO1A） was under-expressed in the SIN-treated PBMCs.Further bioinformatics search indicated that the changes of SNCA,ANP32A and CLIC1 pertained to apoptosis,while changes of GSN and CORO1A were associated with both apoptosis and inhibition of immunological function.Subsequently GSN and CORO1A were selected to validate by Western blotting and the results were consistent with those of 2-DE.Conclusion There were 8 differentially expressed proteins in the SIN-treated PBMCs,which might shed some light on the mechanism of SIN in the treatment of AS.
基金This work was partly supported by a grant from National Science Foundation of China （No.30871122）.
文摘Background Azithromycin can reduce neutrophil accumulation in neutrophilic pulmonary diseases.However,the precise mechanism behind this action remains unknown.Our experiment assessed whether azithromycin inhibits neutrophil accumulation in the airways by affecting interleukin-17 （IL-17） downstream signals.Methods Mice were pretreated with azithromycin before murine IL-17A （mlL-17） stimulation.After the mlL-17 stimulation,the levels of six neutrophil-mobilizing cytokines were determined by enzyme-linked immunosorbent assay （ELISA） tests in bronchoalveolar lavage （BAL） fluid; IL-6,CXC chemokine ligand-1 （CXCL-1）,CXCL-5,macrophage inflammatory protein-2 （MIP-2）,granulocyte colony-stimulating factor （G-CSF）,and granulocyte macrophage colony-stimulating factor （GM-CSF）.The number of neutrophils in BAL fluid were evaluated by cytospin preparations.Results （1） Azithromycin pretreatment significantly inhibited both the release of three neutrophil-mobilizing cytokines （MIP-2,CXCL-5 and GM-CSF） and the accumulation of neutrophils in airways caused by mlL-17 stimulation.（2） The levels of three neutrophil-mobilizing cytokines （IL-6,MIP-2 and GM-CSF） were positively correlated with the numbers of neutrophil in BAL fluid.Conclusions Azithromycin can inhibit neutrophil accumulation in the airways by affecting IL-17 downstream signals.This finding suggests that macrolide antibiotic application might be useful in prevention of neutrophilic pulmonary diseases characterized by high levels of IL-17.
