Objective:Immune dysfunction plays an important role in the pathogenesis of endometriosis.C5a,a by-product of complement activation,binds to its receptor(C5a receptor 1,C5aR1)in various cells,including macrophages.The...Objective:Immune dysfunction plays an important role in the pathogenesis of endometriosis.C5a,a by-product of complement activation,binds to its receptor(C5a receptor 1,C5aR1)in various cells,including macrophages.The combination of C5a and C5aR1 in macrophages induces M2 polarization.There is no consistent conclusion regarding macrophage polarization status and its participation in the pathogenesis of endometriosis.Methods:We analyzed the expression of C5aR1 and the immunophenotype of macrophages in samples from patients with endometriosis using immunohistochemistry and flow cytometry.We tested the equivalent item in a mouse model and the THP-1 cell line in vivo and in vitro.Single-cell RNA sequencing(scRNA-seq)from the Gene Expression Omnibus(GEO,GSE213216)was used to investigate the phenotype and function of C5aR1^(+)macrophages.Results:The infiltration of C5aR1^(+)macrophages was associated with the severity of endometriosis.C5aR1^(+)macrophages polarized more toward the M2 phenotype and transformed into an immunosuppressive state.Inhibiting C5aR1 using PMX-53 decreased the expression of C5aR1^(+)macrophages and reduced their polarization to the M2 phenotype,consequently shrinking lesion size in a mouse model.Conclusion:Our study emphasizes the importance of C5aR1 and the macrophage phenotype in the pathogenesis of endometriosis.We propose PMX-53 as a potential therapeutic agent for endometriosis.展开更多
In the search for a therapeutic schedule for spinal cord injury,it is necessary to understand key genes and their corresponding regulatory networks involved in the spinal cord injury process.However,ad hoc selection a...In the search for a therapeutic schedule for spinal cord injury,it is necessary to understand key genes and their corresponding regulatory networks involved in the spinal cord injury process.However,ad hoc selection and analysis of one or two genes cannot fully reveal the complex molecular biological mechanisms of spinal cord injury.The emergence of second-generation sequencing technology(RNA sequencing)has provided a better method.In this study,RNA sequencing technology was used to analyze differentially expressed genes at different time points after spinal cord injury in rat models established by contusion of the eighth thoracic segment.The numbers of genes that changed significantly were 944,1362 and 1421 at 1,4 and 7 days after spinal cord injury respectively.After gene ontology analysis and temporal expression analysis of the differentially expressed genes,C5ar1,Socs3 and CCL6 genes were then selected and identified by real-time polymerase chain reaction and western blot assay.The mRNA expression trends of C5ar1,Socs3 and CCL6 genes were consistent with the RNA sequencing results.Further verification and analysis of C5ar1 indicate that the level of protein expression of C5ar1 was consistent with its nucleic acid level after spinal cord injury.C5ar1 was mainly expressed in neurons and astrocytes.Finally,the gene Itgb2,which may be related to C5ar1,was found by Chilibot database and literature search.Immunofluorescence histochemical results showed that the expression of Itgb2 was highly consistent with that of C5ar1.Itgb2 was expressed in astrocytes.RNA sequencing technology can screen differentially expressed genes at different time points after spinal cord injury.Through analysis and verification,genes strongly associated with spinal cord injury can be screened.This can provide experimental data for further determining the molecular mechanism of spinal cord injury,and also provide possible targets for the treatment of spinal cord injury.This study was approved ethically by the Laboratory Animal Ethics Committee of Jiangsu Province,China(approval No.2018-0306-001)on March 6,2018.展开更多
基金supported by the Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases,the Obstetrics and Gynecology Hospital,Fudan University。
文摘Objective:Immune dysfunction plays an important role in the pathogenesis of endometriosis.C5a,a by-product of complement activation,binds to its receptor(C5a receptor 1,C5aR1)in various cells,including macrophages.The combination of C5a and C5aR1 in macrophages induces M2 polarization.There is no consistent conclusion regarding macrophage polarization status and its participation in the pathogenesis of endometriosis.Methods:We analyzed the expression of C5aR1 and the immunophenotype of macrophages in samples from patients with endometriosis using immunohistochemistry and flow cytometry.We tested the equivalent item in a mouse model and the THP-1 cell line in vivo and in vitro.Single-cell RNA sequencing(scRNA-seq)from the Gene Expression Omnibus(GEO,GSE213216)was used to investigate the phenotype and function of C5aR1^(+)macrophages.Results:The infiltration of C5aR1^(+)macrophages was associated with the severity of endometriosis.C5aR1^(+)macrophages polarized more toward the M2 phenotype and transformed into an immunosuppressive state.Inhibiting C5aR1 using PMX-53 decreased the expression of C5aR1^(+)macrophages and reduced their polarization to the M2 phenotype,consequently shrinking lesion size in a mouse model.Conclusion:Our study emphasizes the importance of C5aR1 and the macrophage phenotype in the pathogenesis of endometriosis.We propose PMX-53 as a potential therapeutic agent for endometriosis.
基金supported by the National Natural Science Foundation of China,No.31570983(to XDW)the Priority Academic Program Development of Jiangsu Higher Education Institutes of China
文摘In the search for a therapeutic schedule for spinal cord injury,it is necessary to understand key genes and their corresponding regulatory networks involved in the spinal cord injury process.However,ad hoc selection and analysis of one or two genes cannot fully reveal the complex molecular biological mechanisms of spinal cord injury.The emergence of second-generation sequencing technology(RNA sequencing)has provided a better method.In this study,RNA sequencing technology was used to analyze differentially expressed genes at different time points after spinal cord injury in rat models established by contusion of the eighth thoracic segment.The numbers of genes that changed significantly were 944,1362 and 1421 at 1,4 and 7 days after spinal cord injury respectively.After gene ontology analysis and temporal expression analysis of the differentially expressed genes,C5ar1,Socs3 and CCL6 genes were then selected and identified by real-time polymerase chain reaction and western blot assay.The mRNA expression trends of C5ar1,Socs3 and CCL6 genes were consistent with the RNA sequencing results.Further verification and analysis of C5ar1 indicate that the level of protein expression of C5ar1 was consistent with its nucleic acid level after spinal cord injury.C5ar1 was mainly expressed in neurons and astrocytes.Finally,the gene Itgb2,which may be related to C5ar1,was found by Chilibot database and literature search.Immunofluorescence histochemical results showed that the expression of Itgb2 was highly consistent with that of C5ar1.Itgb2 was expressed in astrocytes.RNA sequencing technology can screen differentially expressed genes at different time points after spinal cord injury.Through analysis and verification,genes strongly associated with spinal cord injury can be screened.This can provide experimental data for further determining the molecular mechanism of spinal cord injury,and also provide possible targets for the treatment of spinal cord injury.This study was approved ethically by the Laboratory Animal Ethics Committee of Jiangsu Province,China(approval No.2018-0306-001)on March 6,2018.
