目的探讨全反式维甲酸(ATRA)调控Kruppel因子5(Klf5)在预防移植静脉术后狭窄的作用及机制。方法建立新西兰大白兔(海军军医大学动物实验中心提供)静脉动脉化动物模型,按完全随机分为造模组(A、B、C组分别为饲养2、4、8周)和ATRA组(D、E...目的探讨全反式维甲酸(ATRA)调控Kruppel因子5(Klf5)在预防移植静脉术后狭窄的作用及机制。方法建立新西兰大白兔(海军军医大学动物实验中心提供)静脉动脉化动物模型,按完全随机分为造模组(A、B、C组分别为饲养2、4、8周)和ATRA组(D、E、F组分别为饲养2、4、8周,鼻饲ATRA10 mg/d)、空白对照组(G组),各6只。分别获取移植静脉标本,行苏木精-伊红染色及免疫组化检验。建立人脐静脉平滑肌细胞(SMC)KLF5过表达细胞模型,以ATRA干预,划痕实验、细胞增殖实验(CCK-8)明确SMC增殖及迁移情况;免疫共沉淀明确ATRA对Klf5-RARa结合的阻断作用。两组间计量资料采用两独立样本t检验,多组间计量资料采用单因素方差分析,组间两两比较采用LSD-t检验方法。结果各组管径:A组(107.58±18.11)μm,B组(144.65±26.10)μm,C组(160.28±28.06)μm,D组(78.42±9.00)μm,E组(102.75±16.47)μm,F组(117.47±38.06)μm,G组(35.73±6.04)μm。组间比较,建模组各组(A、B、C各组)明显高于空白对照组(G组)(t=5.81、8.81、10.08,P<0.01),同期建模组各组明显高于ATRA组(t=2.06、2.96、3.03,P<0.05)。以染色指数法计算免疫组化结果,建模各组细胞核增殖抗原(Ki-67,2周3.07±0.64,4周3.67±0.81,8周1.93±0.41)表达明显高于对照组(t=6.93、9.37、4.16,P<0.01),也高于同期ATRA组(2周2.87±0.52,4周3.60±1.21,8周2.10±0.24,t=4.91、6.66、2.14,P<0.05)。实验组Klf5表达(2周4.43±0.70,4周5.67±1.18,8周3.03±0.98)明显高于对照组(t=7.27、9.09、3.83,P<0.01),建模组与ATRA组间KLF5(2周4.43±0.70,4周5.67±1.18,8周3.03±0.98)表达差异无统计学意义(t=0.49、0.17、0.42,P>0.05)。CCK-8实验:72 h Klf5组吸光度(A)值(1.54±0.20)高于其余3组(t=5.62、5.84、8.31,P<0.01),Klf5+ATRA组(1.02±0.14)高于ARTA组(0.80±0.07,t=2.47,P<0.05),对照组(1.04±0.13)高于ATRA组(t=2.70,P<0.05)。划痕实验:以迁移前后划痕距离的比值,klf5组0.098±0.006,对照组0.404±0.009,ATRA组0.597±0.014,Klf5展开更多
【目的】研究Kruppel样转录因子5(Kruppel like factors 5,KLF5)在胰腺癌组织中的表达情况和它的表达与疾病临床病理特征的关系,评价其对胰腺癌患者预后的影响,并分析KLF5在胰腺癌发生、发展中可能的病理机制。【方法】收集整理GEO与TCG...【目的】研究Kruppel样转录因子5(Kruppel like factors 5,KLF5)在胰腺癌组织中的表达情况和它的表达与疾病临床病理特征的关系,评价其对胰腺癌患者预后的影响,并分析KLF5在胰腺癌发生、发展中可能的病理机制。【方法】收集整理GEO与TCGA数据库资料;分析KLF5在胰腺癌与正常胰腺组织表达情况的差异,利用t检验统计KLF5的表达与胰腺癌临床病理特征之间的关系;采用基因富集分析法(gene set enrichment analysis,GSEA)预测KLF5相关的基因通路。【结果】在GSE28735数据集中,KLF5在胰腺癌中的表达明显高于正常的胰腺组织,KLF5的表达与肿瘤的分级、TNM分期存在相关性(P<0.05);在TCGA数据集中,高表达KLF5的胰腺癌患者总体生存率明显低于低表达组(P=0.002);GESA结果显示,KLF5可能富集到E2F通路、G2/M检查点、MYC-V1通路、MYC-V2通路、MTORC1通路、有丝分裂通路、DNA修复通路、糖酵解通路相关的基因集。【结论】胰腺癌组织中KLF5的表达高于正常胰腺组织,KLF5的表达与胰腺癌的分级、TNM分期有关,高表达KLF5的胰腺癌患者临床预后不良,有可能作为胰腺癌患者肿瘤进展的标志物和治疗胰腺癌的新的分子靶标。展开更多
Chronic inflammation plays a pivotal role in insulin resistance and type 2 diabetes,yet the mechanisms are not completely understood.Here,we demonstrated that serum LPS levels were significantly higher in newly diagno...Chronic inflammation plays a pivotal role in insulin resistance and type 2 diabetes,yet the mechanisms are not completely understood.Here,we demonstrated that serum LPS levels were significantly higher in newly diagnosed diabetic patients than in normal control.miR-145 level in peripheral blood mononuclear cells decreased in type 2 diabetics.LPS repressed the transcription of miR-143/145 cluster and decreased miR-145 levels.Attenuation of miR-145 activity by anti-miR-145 triggered liver inflammation and increased serum chemokines in C57BL/6 J mice.Conversely,lentivirus-mediated miR-145 overexpression inhibited macrophage infiltration,reduced body weight,and improved glucose metabolism in db/db mice.And miR-145 overexpression markedly reduced plaque size in the aorta in ApoE−/−mice.Both OPG and KLF5 were targets of miR-145.miR-145 repressed cell proliferation and induced apoptosis partially by targeting OPG and KLF5.miR-145 also suppressed NF-κB activation by targeting OPG and KLF5.Our findings provide an association of the environment with the progress of metabolic disorders.Increasing miR-145 may be a new potential therapeutic strategy in preventing and treating metabolic diseases such as type 2 diabetes and atherosclerosis.展开更多
Background:Accumulated studies have demonstrated that Kruppel‑like factor 5(KLF5),a transcription factor,plays an important role in regulating cell proliferation and tissue remodeling through the expression of its dow...Background:Accumulated studies have demonstrated that Kruppel‑like factor 5(KLF5),a transcription factor,plays an important role in regulating cell proliferation and tissue remodeling through the expression of its downstream genes.KLF5‑related factors are expected to be involved in the healing process after myocardial injury or myocardial ischemic changes,especially for the forensic diagnosis of myocardial ischemic physiopathology.Aim and Objectives:This study aimed to explore the discrimination ability and applicability of KLF5-related factors in SCD caused by MI compared with other causes of death to provide further insights into the forensic diagnosis of myocardial ischemic pathology.Materials and Methods:The relative quantification of F‑Box and WD Repeat Domain Containing 7(FBW7),KLF5,factor‑binding protein(FGFBP)1,and FGFBP2 messenger RNAs(mRNAs)in myocardial tissue samples was performed using real‑time fluorescence quantitative reverse transcription polymerase chain reaction.KLF5 and FGFBP1/2 protein levels were examined using immunohistochemistry(IHC).The forensic autopsy cases(27 in total,autopsy within 72 h postmortem)included seven cases of acute myocardial infarction and 10 cases of acute myocardial ischemia.There were 10 cases in the control group,including four cases of traffic injury one case of injury by fall from height,one case of electric death,and four cases of blunt force injury.Results:Characteristic results were found in myocardial samples from three groups of deaths:KLF5 and FGFBP1 mRNA levels were significantly elevated in the infarction and ischemia groups,while FBW7 mRNA levels were significantly decreased.FBW7 is an important ubiquitin ligase that can mediate the degradation of KLF5 protein.In addition,FBW7 and FGFBP2 mRNA levels were decreased in the infarction group compared with the ischemia group.The IHC results were consistent with the observed mRNA expression patterns.Conclusions:Quantitative detection of FBW7,KLF5,FGFBP1,and FGFBP2 mRNA transcripts in myocardial tissues展开更多
文摘目的探讨全反式维甲酸(ATRA)调控Kruppel因子5(Klf5)在预防移植静脉术后狭窄的作用及机制。方法建立新西兰大白兔(海军军医大学动物实验中心提供)静脉动脉化动物模型,按完全随机分为造模组(A、B、C组分别为饲养2、4、8周)和ATRA组(D、E、F组分别为饲养2、4、8周,鼻饲ATRA10 mg/d)、空白对照组(G组),各6只。分别获取移植静脉标本,行苏木精-伊红染色及免疫组化检验。建立人脐静脉平滑肌细胞(SMC)KLF5过表达细胞模型,以ATRA干预,划痕实验、细胞增殖实验(CCK-8)明确SMC增殖及迁移情况;免疫共沉淀明确ATRA对Klf5-RARa结合的阻断作用。两组间计量资料采用两独立样本t检验,多组间计量资料采用单因素方差分析,组间两两比较采用LSD-t检验方法。结果各组管径:A组(107.58±18.11)μm,B组(144.65±26.10)μm,C组(160.28±28.06)μm,D组(78.42±9.00)μm,E组(102.75±16.47)μm,F组(117.47±38.06)μm,G组(35.73±6.04)μm。组间比较,建模组各组(A、B、C各组)明显高于空白对照组(G组)(t=5.81、8.81、10.08,P<0.01),同期建模组各组明显高于ATRA组(t=2.06、2.96、3.03,P<0.05)。以染色指数法计算免疫组化结果,建模各组细胞核增殖抗原(Ki-67,2周3.07±0.64,4周3.67±0.81,8周1.93±0.41)表达明显高于对照组(t=6.93、9.37、4.16,P<0.01),也高于同期ATRA组(2周2.87±0.52,4周3.60±1.21,8周2.10±0.24,t=4.91、6.66、2.14,P<0.05)。实验组Klf5表达(2周4.43±0.70,4周5.67±1.18,8周3.03±0.98)明显高于对照组(t=7.27、9.09、3.83,P<0.01),建模组与ATRA组间KLF5(2周4.43±0.70,4周5.67±1.18,8周3.03±0.98)表达差异无统计学意义(t=0.49、0.17、0.42,P>0.05)。CCK-8实验:72 h Klf5组吸光度(A)值(1.54±0.20)高于其余3组(t=5.62、5.84、8.31,P<0.01),Klf5+ATRA组(1.02±0.14)高于ARTA组(0.80±0.07,t=2.47,P<0.05),对照组(1.04±0.13)高于ATRA组(t=2.70,P<0.05)。划痕实验:以迁移前后划痕距离的比值,klf5组0.098±0.006,对照组0.404±0.009,ATRA组0.597±0.014,Klf5
文摘【目的】研究Kruppel样转录因子5(Kruppel like factors 5,KLF5)在胰腺癌组织中的表达情况和它的表达与疾病临床病理特征的关系,评价其对胰腺癌患者预后的影响,并分析KLF5在胰腺癌发生、发展中可能的病理机制。【方法】收集整理GEO与TCGA数据库资料;分析KLF5在胰腺癌与正常胰腺组织表达情况的差异,利用t检验统计KLF5的表达与胰腺癌临床病理特征之间的关系;采用基因富集分析法(gene set enrichment analysis,GSEA)预测KLF5相关的基因通路。【结果】在GSE28735数据集中,KLF5在胰腺癌中的表达明显高于正常的胰腺组织,KLF5的表达与肿瘤的分级、TNM分期存在相关性(P<0.05);在TCGA数据集中,高表达KLF5的胰腺癌患者总体生存率明显低于低表达组(P=0.002);GESA结果显示,KLF5可能富集到E2F通路、G2/M检查点、MYC-V1通路、MYC-V2通路、MTORC1通路、有丝分裂通路、DNA修复通路、糖酵解通路相关的基因集。【结论】胰腺癌组织中KLF5的表达高于正常胰腺组织,KLF5的表达与胰腺癌的分级、TNM分期有关,高表达KLF5的胰腺癌患者临床预后不良,有可能作为胰腺癌患者肿瘤进展的标志物和治疗胰腺癌的新的分子靶标。
基金This work was supported by grants from the National Natural Science Foundation of China(81270902,81381220308,and 30230380).
文摘Chronic inflammation plays a pivotal role in insulin resistance and type 2 diabetes,yet the mechanisms are not completely understood.Here,we demonstrated that serum LPS levels were significantly higher in newly diagnosed diabetic patients than in normal control.miR-145 level in peripheral blood mononuclear cells decreased in type 2 diabetics.LPS repressed the transcription of miR-143/145 cluster and decreased miR-145 levels.Attenuation of miR-145 activity by anti-miR-145 triggered liver inflammation and increased serum chemokines in C57BL/6 J mice.Conversely,lentivirus-mediated miR-145 overexpression inhibited macrophage infiltration,reduced body weight,and improved glucose metabolism in db/db mice.And miR-145 overexpression markedly reduced plaque size in the aorta in ApoE−/−mice.Both OPG and KLF5 were targets of miR-145.miR-145 repressed cell proliferation and induced apoptosis partially by targeting OPG and KLF5.miR-145 also suppressed NF-κB activation by targeting OPG and KLF5.Our findings provide an association of the environment with the progress of metabolic disorders.Increasing miR-145 may be a new potential therapeutic strategy in preventing and treating metabolic diseases such as type 2 diabetes and atherosclerosis.
基金supported by the Beijing Natural Science Foundation(grant number 7192121,China)General Program of National Natural Science Foundation of China(grant number 81971796,China).
文摘Background:Accumulated studies have demonstrated that Kruppel‑like factor 5(KLF5),a transcription factor,plays an important role in regulating cell proliferation and tissue remodeling through the expression of its downstream genes.KLF5‑related factors are expected to be involved in the healing process after myocardial injury or myocardial ischemic changes,especially for the forensic diagnosis of myocardial ischemic physiopathology.Aim and Objectives:This study aimed to explore the discrimination ability and applicability of KLF5-related factors in SCD caused by MI compared with other causes of death to provide further insights into the forensic diagnosis of myocardial ischemic pathology.Materials and Methods:The relative quantification of F‑Box and WD Repeat Domain Containing 7(FBW7),KLF5,factor‑binding protein(FGFBP)1,and FGFBP2 messenger RNAs(mRNAs)in myocardial tissue samples was performed using real‑time fluorescence quantitative reverse transcription polymerase chain reaction.KLF5 and FGFBP1/2 protein levels were examined using immunohistochemistry(IHC).The forensic autopsy cases(27 in total,autopsy within 72 h postmortem)included seven cases of acute myocardial infarction and 10 cases of acute myocardial ischemia.There were 10 cases in the control group,including four cases of traffic injury one case of injury by fall from height,one case of electric death,and four cases of blunt force injury.Results:Characteristic results were found in myocardial samples from three groups of deaths:KLF5 and FGFBP1 mRNA levels were significantly elevated in the infarction and ischemia groups,while FBW7 mRNA levels were significantly decreased.FBW7 is an important ubiquitin ligase that can mediate the degradation of KLF5 protein.In addition,FBW7 and FGFBP2 mRNA levels were decreased in the infarction group compared with the ischemia group.The IHC results were consistent with the observed mRNA expression patterns.Conclusions:Quantitative detection of FBW7,KLF5,FGFBP1,and FGFBP2 mRNA transcripts in myocardial tissues
