Interferon regulatory factor 7 plays a crucial role in the innate immune response.However,whether interferon regulatory factor 7-mediated signaling contributes to Parkinson's disease remains unknown.Here we report...Interferon regulatory factor 7 plays a crucial role in the innate immune response.However,whether interferon regulatory factor 7-mediated signaling contributes to Parkinson's disease remains unknown.Here we report that interferon regulatory factor 7 is markedly up-regulated in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced mouse model of Parkinson's disease and co-localizes with microglial cells.Both the selective cyclic guanosine monophosphate adenosine monophosphate synthase inhibitor RU.521 and the stimulator of interferon genes inhibitor H151 effectively suppressed interferon regulatory factor 7 activation in BV2 microglia exposed to 1-methyl-4-phenylpyridinium and inhibited transformation of mouse BV2 microglia into the neurotoxic M1 phenotype.In addition,si RNA-mediated knockdown of interferon regulatory factor 7 expression in BV2 microglia reduced the expression of inducible nitric oxide synthase,tumor necrosis factorα,CD16,CD32,and CD86 and increased the expression of the anti-inflammatory markers ARG1 and YM1.Taken together,our findings indicate that the cyclic guanosine monophosphate adenosine monophosphate synthase-stimulator of interferon genes-interferon regulatory factor 7 pathway plays a crucial role in the pathogenesis of Parkinson's disease.展开更多
Interferon Regulatory Factor-2 (IRF-2) belongs to IRF family, was identified as a mammalian transcription factor involved in Interferon beta (IFNβ) gene regulation. Besides that IRF-2 is involved in immunomodulation,...Interferon Regulatory Factor-2 (IRF-2) belongs to IRF family, was identified as a mammalian transcription factor involved in Interferon beta (IFNβ) gene regulation. Besides that IRF-2 is involved in immunomodulation, hematopoietic differentiation, cell cycle regulation and oncogenesis. We have done molecular sub-cloning and expression of recombinant murine IRF-2 as GST (Glutathione-S-Transferase)- IRF-2 fusion protein in E. coli/XL-1blue cells. Recombinant IRF-2 with GST moiety at N-terminus expressed as GST-IRF-2 (~66 kd) in E. coli along with different low molecular mass degradation products revealed approximately 30, 42, 60 and 62 kd by SDS-PAGE and Western blot, respectively. We further confirm that degradation takes place at C-terminus of the fusion protein not at N-terminus as anti-GST antibody was detecting all bands in the immunoblot. The recombinant IRF-2 was biologically active along with their degradation products in terms of their DNA binding activity as assessed by Electrophoretically Mobility Shift Assay (EMSA). We observed three different molecular mass DNA/protein complexes (1 - 3) with Virus Response Element (VRE) derived from human Interferon IFNβ gene and five different molecular mass complexes (1 - 5) with IRF-E motif (GAAAGT)4 in EMSA gel. GST only expressed from empty vector did not bind to these DNA elements. To confirm that the binding is specific, all complexes were competed out completely when challenged with 100-X fold molar excess of IRF-E oligo under cold competition. It means degradation products along with full-length protein are able to interact with VREβ as well as IRF-E motif. This means degradation products may regulate the target gene (s) activation/repression via interacting with VRE/IRF-E.展开更多
Heart diseases are the main cause of mortality in Mexico, being coronary </span><span style="font-family:Verdana;">heart disease the most frequent in the country. Its high prevalence makes i...Heart diseases are the main cause of mortality in Mexico, being coronary </span><span style="font-family:Verdana;">heart disease the most frequent in the country. Its high prevalence makes important </span><span style="font-family:Verdana;">the study of the pathophysiology and the search for prognostic </span><span style="font-family:Verdana;">factors. Different genes and polymorphisms promote atherogenesis and coronary artery disease, they affect inflammatory and vascular pathological processes. </span><span style="font-family:Verdana;">Interferon regulatory factor 5 (IRF5) is associated with coronary heart disease, it promotes chronic inflammation and cytokines release;it could trigger immune reactions and its activating receptors express in the vascular endothelium. Besides, polymorphisms in the renin-angiotensin-aldosterone system (RAAS) are implied with coronary disease, they are found in angiotensinogen (AGT), angiotensin II type 1 receptor (AT1R), angiotensin II type 2 receptor (AT2R), and angiotensin-converting enzyme (ACE) genes. These genetic polymorphisms are associated with a prothrombotic state, endothelial dysfunction, and immune activation. Multiple experimental studies showed that chronic activation of RAAS and chronic expression of IRF5 generates an environment prone to the development of atherosclerosis, and autoimmune and cardiovascular diseases. Studying these specific genes and their relationship with coronary heart disease will allow a better understanding of the pathological process and possibly the quest for new treatments.展开更多
目的探讨骨形成蛋白7(BMP7)治疗葡聚糖硫酸钠(DSS)诱导的大鼠结肠炎症的疗效及其机制。方法健康雄性大鼠60只,随机平均分为正常对照组、DSS组(DSS诱导结肠炎症组)及DSS+BMP7组(BMP7治疗DSS诱导的结肠炎症组),每组20只。正常对照组大鼠...目的探讨骨形成蛋白7(BMP7)治疗葡聚糖硫酸钠(DSS)诱导的大鼠结肠炎症的疗效及其机制。方法健康雄性大鼠60只,随机平均分为正常对照组、DSS组(DSS诱导结肠炎症组)及DSS+BMP7组(BMP7治疗DSS诱导的结肠炎症组),每组20只。正常对照组大鼠饮用灭菌水,DSS组及DSS+BMP7组大鼠每天饮用3.5%的DSS以建立肠炎模型,DSS+BMP7组隔日1次腹腔注射1 m L BMP7(0.1μg/m L)。于第0、7、14及28天各组分别处死5只大鼠,采用组织学评分方法判断炎症程度,并于第28天采用流式细胞术检测外周血中调节性T细胞(Treg)比例变化,切取结肠组织分别利用实时PCR及Western blot检测Foxp3m RNA及蛋白的表达,采用ELISA法检测血清中转化生长因子β1(TGF-β1)水平。结果第7天时炎症程度DSS+BMP7组和DSS组低于正常对照组(P<0.05),但DSS+BMP7组与DSS组相比无统计学差异(P>0.05);第14和28天时DSS+BMP7组炎症程度低于DSS组(P<0.05),第28天时DSS+BMP7组与正常对照组相比无统计学差异(P>0.05)。第28天时Treg细胞比例DSS组及DSS+BMP7组低于正常对照组,且DSS+BMP7组显著高于DSS组(P>0.05);与正常对照组相比,DSS组和DSS+BMP7组中Foxp3的表达降低,且DSS+BMP7组高于DSS组(P<0.05)。DSS组和DSS+BMP7组细胞因子TGF-β1水平低于正常对照组,且DSS+BMP7组高于DSS组(P<0.05)。结论 BMP7可能作用于Treg细胞,减轻DSS诱导的结肠炎性疾病。展开更多
基金supported by the National Natural Science Foundation of China,Nos.82171429,81771384a grant from Wuxi Municipal Health Commission,No.1286010241190480(all to YS)。
文摘Interferon regulatory factor 7 plays a crucial role in the innate immune response.However,whether interferon regulatory factor 7-mediated signaling contributes to Parkinson's disease remains unknown.Here we report that interferon regulatory factor 7 is markedly up-regulated in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced mouse model of Parkinson's disease and co-localizes with microglial cells.Both the selective cyclic guanosine monophosphate adenosine monophosphate synthase inhibitor RU.521 and the stimulator of interferon genes inhibitor H151 effectively suppressed interferon regulatory factor 7 activation in BV2 microglia exposed to 1-methyl-4-phenylpyridinium and inhibited transformation of mouse BV2 microglia into the neurotoxic M1 phenotype.In addition,si RNA-mediated knockdown of interferon regulatory factor 7 expression in BV2 microglia reduced the expression of inducible nitric oxide synthase,tumor necrosis factorα,CD16,CD32,and CD86 and increased the expression of the anti-inflammatory markers ARG1 and YM1.Taken together,our findings indicate that the cyclic guanosine monophosphate adenosine monophosphate synthase-stimulator of interferon genes-interferon regulatory factor 7 pathway plays a crucial role in the pathogenesis of Parkinson's disease.
文摘Interferon Regulatory Factor-2 (IRF-2) belongs to IRF family, was identified as a mammalian transcription factor involved in Interferon beta (IFNβ) gene regulation. Besides that IRF-2 is involved in immunomodulation, hematopoietic differentiation, cell cycle regulation and oncogenesis. We have done molecular sub-cloning and expression of recombinant murine IRF-2 as GST (Glutathione-S-Transferase)- IRF-2 fusion protein in E. coli/XL-1blue cells. Recombinant IRF-2 with GST moiety at N-terminus expressed as GST-IRF-2 (~66 kd) in E. coli along with different low molecular mass degradation products revealed approximately 30, 42, 60 and 62 kd by SDS-PAGE and Western blot, respectively. We further confirm that degradation takes place at C-terminus of the fusion protein not at N-terminus as anti-GST antibody was detecting all bands in the immunoblot. The recombinant IRF-2 was biologically active along with their degradation products in terms of their DNA binding activity as assessed by Electrophoretically Mobility Shift Assay (EMSA). We observed three different molecular mass DNA/protein complexes (1 - 3) with Virus Response Element (VRE) derived from human Interferon IFNβ gene and five different molecular mass complexes (1 - 5) with IRF-E motif (GAAAGT)4 in EMSA gel. GST only expressed from empty vector did not bind to these DNA elements. To confirm that the binding is specific, all complexes were competed out completely when challenged with 100-X fold molar excess of IRF-E oligo under cold competition. It means degradation products along with full-length protein are able to interact with VREβ as well as IRF-E motif. This means degradation products may regulate the target gene (s) activation/repression via interacting with VRE/IRF-E.
文摘Heart diseases are the main cause of mortality in Mexico, being coronary </span><span style="font-family:Verdana;">heart disease the most frequent in the country. Its high prevalence makes important </span><span style="font-family:Verdana;">the study of the pathophysiology and the search for prognostic </span><span style="font-family:Verdana;">factors. Different genes and polymorphisms promote atherogenesis and coronary artery disease, they affect inflammatory and vascular pathological processes. </span><span style="font-family:Verdana;">Interferon regulatory factor 5 (IRF5) is associated with coronary heart disease, it promotes chronic inflammation and cytokines release;it could trigger immune reactions and its activating receptors express in the vascular endothelium. Besides, polymorphisms in the renin-angiotensin-aldosterone system (RAAS) are implied with coronary disease, they are found in angiotensinogen (AGT), angiotensin II type 1 receptor (AT1R), angiotensin II type 2 receptor (AT2R), and angiotensin-converting enzyme (ACE) genes. These genetic polymorphisms are associated with a prothrombotic state, endothelial dysfunction, and immune activation. Multiple experimental studies showed that chronic activation of RAAS and chronic expression of IRF5 generates an environment prone to the development of atherosclerosis, and autoimmune and cardiovascular diseases. Studying these specific genes and their relationship with coronary heart disease will allow a better understanding of the pathological process and possibly the quest for new treatments.
文摘目的探讨骨形成蛋白7(BMP7)治疗葡聚糖硫酸钠(DSS)诱导的大鼠结肠炎症的疗效及其机制。方法健康雄性大鼠60只,随机平均分为正常对照组、DSS组(DSS诱导结肠炎症组)及DSS+BMP7组(BMP7治疗DSS诱导的结肠炎症组),每组20只。正常对照组大鼠饮用灭菌水,DSS组及DSS+BMP7组大鼠每天饮用3.5%的DSS以建立肠炎模型,DSS+BMP7组隔日1次腹腔注射1 m L BMP7(0.1μg/m L)。于第0、7、14及28天各组分别处死5只大鼠,采用组织学评分方法判断炎症程度,并于第28天采用流式细胞术检测外周血中调节性T细胞(Treg)比例变化,切取结肠组织分别利用实时PCR及Western blot检测Foxp3m RNA及蛋白的表达,采用ELISA法检测血清中转化生长因子β1(TGF-β1)水平。结果第7天时炎症程度DSS+BMP7组和DSS组低于正常对照组(P<0.05),但DSS+BMP7组与DSS组相比无统计学差异(P>0.05);第14和28天时DSS+BMP7组炎症程度低于DSS组(P<0.05),第28天时DSS+BMP7组与正常对照组相比无统计学差异(P>0.05)。第28天时Treg细胞比例DSS组及DSS+BMP7组低于正常对照组,且DSS+BMP7组显著高于DSS组(P>0.05);与正常对照组相比,DSS组和DSS+BMP7组中Foxp3的表达降低,且DSS+BMP7组高于DSS组(P<0.05)。DSS组和DSS+BMP7组细胞因子TGF-β1水平低于正常对照组,且DSS+BMP7组高于DSS组(P<0.05)。结论 BMP7可能作用于Treg细胞,减轻DSS诱导的结肠炎性疾病。
