Mammalian macrophage migration inhibitory factor （MIF） plays an important role as an indispensable mediator in the pathogenesis of inflammatory disease like septicemia, but little is known about the role of MIF homo...Mammalian macrophage migration inhibitory factor （MIF） plays an important role as an indispensable mediator in the pathogenesis of inflammatory disease like septicemia, but little is known about the role of MIF homologue in fish septicemia. The authors have cloned the MIF homologue in large yellow croaker Pseudosciaena crocea （LycMIF） using RACE approach. The full-length cDNA of LycMIF was 634 bases and contained an ORF of 345 bases encoding a protein of 115 amino acid residues. As demonstrated by RT-PCR and QRT-PCR assay, MIF mRNAs were constitutively expressed in 11 selected tissues and were abundant in brain and liver. Moreover, the LycMIF transcripts in the liver and head kidney were responsive to bacteria infection and could be significantly up-regulated. Our results provide the first direct evidence that fish MIF was implicated in pathogenesis of fish vibrosis and play an important role in response to bacteria infection.展开更多
Aims T-type Ca<sup>2+</sup> current(I<sub>CaT</sub>)plays an important role in the pathogenesis of atrial fibrillation（AF）.The present study sought to investigate the role of Macrophage migra...Aims T-type Ca<sup>2+</sup> current(I<sub>CaT</sub>)plays an important role in the pathogenesis of atrial fibrillation（AF）.The present study sought to investigate the role of Macrophage migration inhibitory factor（MIF）,a pleiotropic cytokine,in the regulation of T-type Ca<sup>2+</sup> channel in atrium myocytes.Methods We used whole-cell voltage-clamp technique and biochemical assays to study the regulation and expression of I<sub>Ca</sub>,T in mouse atrium myocytes（HL-1 cells）.Results Serum MIF concentrations was slightly increased in patients with AF compared to sinus rhythm（SR） controls.In cultured HL-1 cells, significant amounts of MIF were produced in response to hydrogen peroxide（H<sub>2</sub>O<sub>2</sub>）,but not AngiotensinⅡ（AngⅡ）. Mouse recombinant MIF（rMIF）（20 or 40 nM,24 h） suppressed peak ICa,T by-38%and-60%in a concentration-dependent manner,impaired the voltage-dependent activation of I<sub>Ca</sub>,T,and down-regulated of TCC alG mRNA.Src inhibitors genistein and PPl significantly enhanced ICaT.The depression of ICa,T induced by rMIF could be reversed by genistein and PP1.Conclusions MIFis involved in the pathogenesis of AF,probably by decreasing ICa,T through impairment of the channel function and activation of c-Src kinases in atrium myocytes.展开更多
To investigate the effects of antisense oligonucleotides on the expression of macrophage migration inhibitory factor (MIF) on macrophages, the mouse phosphorothioate oligonucleotides were designed and synthesized with...To investigate the effects of antisense oligonucleotides on the expression of macrophage migration inhibitory factor (MIF) on macrophages, the mouse phosphorothioate oligonucleotides were designed and synthesized with the sequences of antisense, 5′-TACGGATACAAGTAGCAC-3′; Sense, 5′-ATGCCTATGTTCATCGTG-3′; Missense, 5′-CTCTCAGACTCGATCTGT-3′. These phosphorothioate oligonucleotides were then transfected into cultured macrophages ( RAW264.7 ) by luciferase vector, and the transfected macrophages were incubated with Lipopolysaccharide (LPS) (1?ng/ml) for various periods of times and collected afterwards. The content of MIF protein in the cultural supernatants was determined by ELISA, cellular RNA extracted and the expression of MIF mRNA was examined by RT-PCR analysis. The experimental results showed that LPS could induce a time-dependent specific expression of MIF on macrophages, in which the MIF mRNA in cells and the MIF protein in cultural supernatants appeared after 3 h and reached their highest concentration at 9-12?h after LPS stimulation. The levels of mRNA and proteins in the macrophages treated with antisense olignucleotides were decreased significantly after stimulation with LPS in comparison with that of stimulation with LPS alone or with that with LPS plus sense or missense oligonucleotides. There were no differences among those without LPS stimulation. It is concluded that macrophages stimulated with LPS express MIF, and the antisense olignucleotides of MIF inhibit the expression of MIF mRNA as well as the secretion of MIF proteins in macrophages.展开更多
Background:Previous studies have shown that macrophage migration inhibitory factor(MIF)is involved in the pathogenesis of asthma.This study aimed to investigate whether serum MIF reflects a therapeutic response in all...Background:Previous studies have shown that macrophage migration inhibitory factor(MIF)is involved in the pathogenesis of asthma.This study aimed to investigate whether serum MIF reflects a therapeutic response in allergic asthma.Methods:We enrolled 30 asthmatic patients with mild-to-moderate exacerbations and 20 healthy controls,analyzing the parameter levels of serum MIF,serum total immunoglobulin E(tIgE),peripheral blood eosinophil percentage(EOS%),and fractional exhaled nitric oxide(FeNO).Lung function indices were used to identify disease severity and therapeutic response.Results:Our study showed that all measured parameters in patients were at higher levels than those of controls.After one week of treatment,most parameter levels decreased significantly except for serum tIgE.Furthermore,we found that serum MIF positively correlated with EOS%as well as FeNO,but negatively correlated with lung function indices.Receiver operator characteristic(ROC)curve analysis indicated that among the parameters,serum MIF exhibited a higher capacity to evaluate therapeutic response.The area under the curve(AUC)of MIF was 0.931,with a sensitivity of 0.967 and a specificity of 0.800.Conclusions:Our results suggested that serum MIF may serve as a potential biomarker for evaluating therapeutic response in allergic asthma with mild-to-moderate exacerbations.展开更多
Objective:This research was performed to explore the effect of macrophage migration inhibitory factor(MIF)on the apoptosis of bone marrow mesenchymal stem cells(BMSCs)in ischemia and hypoxia environments.Methods:The c...Objective:This research was performed to explore the effect of macrophage migration inhibitory factor(MIF)on the apoptosis of bone marrow mesenchymal stem cells(BMSCs)in ischemia and hypoxia environments.Methods:The cell viability of BMSCs incubated under hypoxia/ischemia(H/I)conditions with or without pretreatment with MIF or triglycidyl isocyanurate(TGIC)was detected using cell counting kit-8(CCK-8)analysis.Plasmids containing long noncoding RNA(lncRNA)maternally expressed gene 3(MEG3)orβ-catenin small interfering RNA(siRNA)were used to overexpress or downregulate the corresponding gene,and the p53 signaling pathway was activated by pretreatment with TGIC.The influences of MIF,overexpression of lncRNA MEG3,activation of the p53 signaling pathway,and silencing ofβ-catenin on H/I-induced apoptosis of BMSCs were revealed by western blotting,flow cytometry,and terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)staining.Results:From the results of CCK-8 assay,western blotting,and flow cytometry,pretreatment with MIF significantly decreased the H/I-induced apoptosis of BMSCs.This effect was inhibited when lncRNA MEG3 was overexpressed by plasmids containing MEG3.The p53 signaling pathway was activated by TGIC,andβ-catenin was silenced by siRNA.From western blot results,the expression levels ofβ-catenin in the nucleus and phosphorylated p53(p-p53)were downregulated and upregulated,respectively,when the lncRNA MEG3 was overexpressed.Through flow cytometry,MIF was also shown to significantly alleviate the increased reactive oxygen species(ROS)level of BMSCs caused by H/I.Conclusions:In summary,we conclude that MIF protected BMSCs from H/I-induced apoptosis by downregulating the lncRNA MEG3/p53 signaling pathway,activating the Wnt/β-catenin signaling pathway,and decreasing ROS levels.展开更多
基金The National Natural Science Foundation of China under contract No. 40976096the National Department Public Benefit Research Foundation of China under contract No. 200903029the National High Technology Research and Development Program of China (863 Program) under contract No. 2006AA10A405
文摘Mammalian macrophage migration inhibitory factor （MIF） plays an important role as an indispensable mediator in the pathogenesis of inflammatory disease like septicemia, but little is known about the role of MIF homologue in fish septicemia. The authors have cloned the MIF homologue in large yellow croaker Pseudosciaena crocea （LycMIF） using RACE approach. The full-length cDNA of LycMIF was 634 bases and contained an ORF of 345 bases encoding a protein of 115 amino acid residues. As demonstrated by RT-PCR and QRT-PCR assay, MIF mRNAs were constitutively expressed in 11 selected tissues and were abundant in brain and liver. Moreover, the LycMIF transcripts in the liver and head kidney were responsive to bacteria infection and could be significantly up-regulated. Our results provide the first direct evidence that fish MIF was implicated in pathogenesis of fish vibrosis and play an important role in response to bacteria infection.
文摘Aims T-type Ca<sup>2+</sup> current(I<sub>CaT</sub>)plays an important role in the pathogenesis of atrial fibrillation（AF）.The present study sought to investigate the role of Macrophage migration inhibitory factor（MIF）,a pleiotropic cytokine,in the regulation of T-type Ca<sup>2+</sup> channel in atrium myocytes.Methods We used whole-cell voltage-clamp technique and biochemical assays to study the regulation and expression of I<sub>Ca</sub>,T in mouse atrium myocytes（HL-1 cells）.Results Serum MIF concentrations was slightly increased in patients with AF compared to sinus rhythm（SR） controls.In cultured HL-1 cells, significant amounts of MIF were produced in response to hydrogen peroxide（H<sub>2</sub>O<sub>2</sub>）,but not AngiotensinⅡ（AngⅡ）. Mouse recombinant MIF（rMIF）（20 or 40 nM,24 h） suppressed peak ICa,T by-38%and-60%in a concentration-dependent manner,impaired the voltage-dependent activation of I<sub>Ca</sub>,T,and down-regulated of TCC alG mRNA.Src inhibitors genistein and PPl significantly enhanced ICaT.The depression of ICa,T induced by rMIF could be reversed by genistein and PP1.Conclusions MIFis involved in the pathogenesis of AF,probably by decreasing ICa,T through impairment of the channel function and activation of c-Src kinases in atrium myocytes.
文摘To investigate the effects of antisense oligonucleotides on the expression of macrophage migration inhibitory factor (MIF) on macrophages, the mouse phosphorothioate oligonucleotides were designed and synthesized with the sequences of antisense, 5′-TACGGATACAAGTAGCAC-3′; Sense, 5′-ATGCCTATGTTCATCGTG-3′; Missense, 5′-CTCTCAGACTCGATCTGT-3′. These phosphorothioate oligonucleotides were then transfected into cultured macrophages ( RAW264.7 ) by luciferase vector, and the transfected macrophages were incubated with Lipopolysaccharide (LPS) (1?ng/ml) for various periods of times and collected afterwards. The content of MIF protein in the cultural supernatants was determined by ELISA, cellular RNA extracted and the expression of MIF mRNA was examined by RT-PCR analysis. The experimental results showed that LPS could induce a time-dependent specific expression of MIF on macrophages, in which the MIF mRNA in cells and the MIF protein in cultural supernatants appeared after 3 h and reached their highest concentration at 9-12?h after LPS stimulation. The levels of mRNA and proteins in the macrophages treated with antisense olignucleotides were decreased significantly after stimulation with LPS in comparison with that of stimulation with LPS alone or with that with LPS plus sense or missense oligonucleotides. There were no differences among those without LPS stimulation. It is concluded that macrophages stimulated with LPS express MIF, and the antisense olignucleotides of MIF inhibit the expression of MIF mRNA as well as the secretion of MIF proteins in macrophages.
基金the Henan Province Medical Science and Technology Research Project (No. 2018020737), China。
文摘Background:Previous studies have shown that macrophage migration inhibitory factor(MIF)is involved in the pathogenesis of asthma.This study aimed to investigate whether serum MIF reflects a therapeutic response in allergic asthma.Methods:We enrolled 30 asthmatic patients with mild-to-moderate exacerbations and 20 healthy controls,analyzing the parameter levels of serum MIF,serum total immunoglobulin E(tIgE),peripheral blood eosinophil percentage(EOS%),and fractional exhaled nitric oxide(FeNO).Lung function indices were used to identify disease severity and therapeutic response.Results:Our study showed that all measured parameters in patients were at higher levels than those of controls.After one week of treatment,most parameter levels decreased significantly except for serum tIgE.Furthermore,we found that serum MIF positively correlated with EOS%as well as FeNO,but negatively correlated with lung function indices.Receiver operator characteristic(ROC)curve analysis indicated that among the parameters,serum MIF exhibited a higher capacity to evaluate therapeutic response.The area under the curve(AUC)of MIF was 0.931,with a sensitivity of 0.967 and a specificity of 0.800.Conclusions:Our results suggested that serum MIF may serve as a potential biomarker for evaluating therapeutic response in allergic asthma with mild-to-moderate exacerbations.
基金supported by the National Natural Science Foundation of China(No.81702132)the Zhejiang Provincial Natural Science Foundation of China(No.LY21H060007)+1 种基金the Projects of Medical and Health Technology Program in Zhejiang Province(No.2021KY206)the Wenzhou Public Welfare Scienceand Technology Research Project(Nos.Y20190267 and Y20210436),China.
文摘Objective:This research was performed to explore the effect of macrophage migration inhibitory factor(MIF)on the apoptosis of bone marrow mesenchymal stem cells(BMSCs)in ischemia and hypoxia environments.Methods:The cell viability of BMSCs incubated under hypoxia/ischemia(H/I)conditions with or without pretreatment with MIF or triglycidyl isocyanurate(TGIC)was detected using cell counting kit-8(CCK-8)analysis.Plasmids containing long noncoding RNA(lncRNA)maternally expressed gene 3(MEG3)orβ-catenin small interfering RNA(siRNA)were used to overexpress or downregulate the corresponding gene,and the p53 signaling pathway was activated by pretreatment with TGIC.The influences of MIF,overexpression of lncRNA MEG3,activation of the p53 signaling pathway,and silencing ofβ-catenin on H/I-induced apoptosis of BMSCs were revealed by western blotting,flow cytometry,and terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)staining.Results:From the results of CCK-8 assay,western blotting,and flow cytometry,pretreatment with MIF significantly decreased the H/I-induced apoptosis of BMSCs.This effect was inhibited when lncRNA MEG3 was overexpressed by plasmids containing MEG3.The p53 signaling pathway was activated by TGIC,andβ-catenin was silenced by siRNA.From western blot results,the expression levels ofβ-catenin in the nucleus and phosphorylated p53(p-p53)were downregulated and upregulated,respectively,when the lncRNA MEG3 was overexpressed.Through flow cytometry,MIF was also shown to significantly alleviate the increased reactive oxygen species(ROS)level of BMSCs caused by H/I.Conclusions:In summary,we conclude that MIF protected BMSCs from H/I-induced apoptosis by downregulating the lncRNA MEG3/p53 signaling pathway,activating the Wnt/β-catenin signaling pathway,and decreasing ROS levels.
