目的:克隆并原核表达小鼠高迁移率族蛋白1(high mobility group box1,HMGB1)。方法:从小鼠脾脏细胞中提取总RNA,应用RT-PCR获得小鼠HMGB1cDNA,经PCR扩增小鼠HMGB1基因;通过T-A克隆将该基因构建到PMD-18T载体中,测序鉴定;将该基因插入pET...目的:克隆并原核表达小鼠高迁移率族蛋白1(high mobility group box1,HMGB1)。方法:从小鼠脾脏细胞中提取总RNA,应用RT-PCR获得小鼠HMGB1cDNA,经PCR扩增小鼠HMGB1基因;通过T-A克隆将该基因构建到PMD-18T载体中,测序鉴定;将该基因插入pET-28a(+)表达载体,IPTG诱导后,可表达相对分子质量约30 kDa的蛋白。蛋白质印迹鉴定表达的目的蛋白。结果:经RT-PCR扩增得到648 bp的DNA片段,序列分析显示与基因库中报道的已知序列完全一致;构建了含有HMGB1编码序列的原核表达载体,经诱导表达、蛋白质印迹鉴定,获得相对分子质量约30 kDa的融合蛋白。结论:成功克隆和表达了小鼠HMGB1基因,为HMGB1蛋白的功能试验奠定了基础。展开更多
Fibroblasts are pleomorphic cells that have a multidirectional effect on organ morphogenesis,tissue homeostasis,and immune response.In fibrotic diseases,fibroblasts synthesize large amounts of extracellular matr...Fibroblasts are pleomorphic cells that have a multidirectional effect on organ morphogenesis,tissue homeostasis,and immune response.In fibrotic diseases,fibroblasts synthesize large amounts of extracellular matrix(ECM),leading to scarring and organ failure.Purine-rich boxl(PU.I)is a specific transcription factor of hematopoietic cell and belongs to the E26 transformation specificity(ETS)family.Recently,it was found that the transcription factor PU.I is an important regulatory factor of the profibrotic gene expression program.TGF-J3 had been proved to play an important role in many ocular tissue fibrosis diseases,and up-regulated the expression of PU.I in fibroblasts producing ECM in a Smad-3 dependent manner.We explore the effect of PU.I on fibrosis of different ocular tissues from this perspective.This article reviews the role of PU.l and its effects on fibrosis of ocular tissue and other tissues.展开更多
Objective: Ethyl Pyruvate (EP) has been shown to be an effective anti-inflammatory agent in a variety of model systems. The aim of this study was to investigate the effects of EP on High Mobility Group Box1(HMGB1) gen...Objective: Ethyl Pyruvate (EP) has been shown to be an effective anti-inflammatory agent in a variety of model systems. The aim of this study was to investigate the effects of EP on High Mobility Group Box1(HMGB1) genes expression and the possible mechanisms of EP protecting against acute lung injury induced by sepsis. Methods: Forty Wistar rats were randomly divided into normal controls,sham operation,acute lung injury,and EP treatment (40 mg/kg intra-peritoneally every 6 hrs) groups. At the time points of 24 hours the animals in each group were sacrificed, and the lungs were harvested. Wet/dry lung weight ratio, the protein in the bronchoalveolar lavage fluid(BALF),and pulmonary permeability index(PPI) were determined. The histological morphology of lung was observed under microscope. The expression of HMGB1 mRNA was measured using semi-quantitative RT-PCR. Results: EP treatment decreased wet/dry lung weight ratio, the protein in the BALF,and PPI (P<0.01). The histological morphology of lung injury was ameliorated. EP significantly inhibited the HMGB1 mRNA expression(P<0.01). HMGB1 mRNA expression in lungs positively correlation with wet/dry lung weight ratio, the protein in the BALF,and PPI. Conclusion: EP administered inhibits HMGB1 mRNA expression, and protects the lungs against acute injury induced by sepsis.展开更多
目的探讨高迁移率蛋白1(HMGB1)在脂多糖诱导的急性肺损伤中的作用,以及甲基强的松龙注射剂干预后其在急性肺损伤中的表达变化。方法将54只清洁级成年雄性SD大鼠随机分成3组:1正常对照组(N组):每只大鼠经尾静脉途径给予生理盐水2 m L/kg,...目的探讨高迁移率蛋白1(HMGB1)在脂多糖诱导的急性肺损伤中的作用,以及甲基强的松龙注射剂干预后其在急性肺损伤中的表达变化。方法将54只清洁级成年雄性SD大鼠随机分成3组:1正常对照组(N组):每只大鼠经尾静脉途径给予生理盐水2 m L/kg,1 h后再经尾静脉途径给予生理盐水1 m L/kg;2急性肺损伤组(ALI组):尾静脉注射脂多糖6 mg/kg,1 h后再经尾静脉途径给予生理盐水1 m L/kg;3甲基强的松龙干预组(MPN组):每只大鼠经尾静脉途径给予脂多糖6 mg/kg,1 h后再经尾静脉途径给予甲基强的松龙20 mg/kg。三组分别于注射后12、24、36 h麻醉处死后采集标本,每个时相点取6只大鼠。采用免疫组织化学法和实时定量PCR(Real-Time PCR)法检测各组各时相点大鼠肺组织HMGB1表达水平,同时观察肺组织病理学改变,并测定干湿重比(W/D)。采用SPSS 20.0统计学软件分析,数据处理采用方差分析。结果 N组中大鼠肺组织有少量HMGB1m RNA及其蛋白表达,ALI组和MPN组大鼠肺组织中HMGB1 m RNA及其蛋白表达水平明显高于N组,差异均有统计学意义(均P<0.05),24 h表达达峰值;同一时相点ALI组大鼠肺组织HMGB1 m RNA及其蛋白表达水平高于MPN组,差异有统计学意义(P<0.05)。ALI组大鼠与MPN组不同时相点W/D增高,与N组比较差异有高度统计学意义(P<0.01),MPN组与ALI组比较W/D下降(P<0.05或P<0.01)。与N组比较,ALI组和MPN组组织病理学有不同程度的病理损害,MPN组比ALI组坏死程度轻,炎症细胞浸润少。结论 HMGB1在炎症后期持续性高水平表达,在ALI发生、发展过程中,尤其是ALI后期失控性炎性反应过程中可能发挥重要作用。甲基强的松龙注射剂通过影响HMGB1 m RNA及其蛋白表达,对脂多糖所致的急性肺损伤有一定的保护作用。展开更多
高迁移率族蛋白1(high mobility group box1,HMGB1)是一种高度保守的非组蛋白染色体结合蛋白。在生理条件下,HMGB1参与核内DNA的转录、复制、修复、重组和核小体形成。在病理条件下,大量释放到细胞外的HMGB1作为重要的炎症介质启动和维...高迁移率族蛋白1(high mobility group box1,HMGB1)是一种高度保守的非组蛋白染色体结合蛋白。在生理条件下,HMGB1参与核内DNA的转录、复制、修复、重组和核小体形成。在病理条件下,大量释放到细胞外的HMGB1作为重要的炎症介质启动和维持炎症反应。HMGB1已被证实与脓毒症、创伤性损伤、自身免疫性疾病以及缺血/再灌注损伤等多种疾病的发病机制密切相关。甘草酸(glycyrrhizic acid)是大量存在于甘草的根和根茎中的一种天然三萜皂苷,具有多种药理功能。研究发现甘草酸作为天然的HMGB1抑制剂,通过与HMGB1结合进而阻断HMGB1与其受体的相互作用从而发挥抗炎活性。本文综述近年来甘草酸及其衍生物治疗HMGB1相关炎症性疾病的药理作用研究,为甘草酸及其衍生物的进一步临床应用提供实验基础。展开更多
高迁移率族蛋白B1(high mobility group protein box 1,HMGB1)是一组高度保守的DNA结合蛋白,存在于体内各种细胞的细胞核中,调控着细胞基因的转录。生理状态下HMGB1发挥着核结合蛋白的作用。当释放到细胞间隙后则会起到晚期炎症因...高迁移率族蛋白B1(high mobility group protein box 1,HMGB1)是一组高度保守的DNA结合蛋白,存在于体内各种细胞的细胞核中,调控着细胞基因的转录。生理状态下HMGB1发挥着核结合蛋白的作用。当释放到细胞间隙后则会起到晚期炎症因子的作用。最近的研究提示,HMGB1不仅参与脓毒症、自身免疫性疾病、慢性肝病、恶性肿瘤等的致病过程,还可参与细胞损伤的修复或致病,在多种疾病及组织器官损伤、修复过程中发挥着重要作用。展开更多
基金Supported by the National Natural Science Foundation of China(No.81470633)the Natural Science Grant of the Heilongjiang Province of China(No.H2018035,No.LH2020H040)the Innovation and Development Foundation of the First Affiliated Hospital of Harbin Medical University(No.2018L002).
文摘Fibroblasts are pleomorphic cells that have a multidirectional effect on organ morphogenesis,tissue homeostasis,and immune response.In fibrotic diseases,fibroblasts synthesize large amounts of extracellular matrix(ECM),leading to scarring and organ failure.Purine-rich boxl(PU.I)is a specific transcription factor of hematopoietic cell and belongs to the E26 transformation specificity(ETS)family.Recently,it was found that the transcription factor PU.I is an important regulatory factor of the profibrotic gene expression program.TGF-J3 had been proved to play an important role in many ocular tissue fibrosis diseases,and up-regulated the expression of PU.I in fibroblasts producing ECM in a Smad-3 dependent manner.We explore the effect of PU.I on fibrosis of different ocular tissues from this perspective.This article reviews the role of PU.l and its effects on fibrosis of ocular tissue and other tissues.
文摘Objective: Ethyl Pyruvate (EP) has been shown to be an effective anti-inflammatory agent in a variety of model systems. The aim of this study was to investigate the effects of EP on High Mobility Group Box1(HMGB1) genes expression and the possible mechanisms of EP protecting against acute lung injury induced by sepsis. Methods: Forty Wistar rats were randomly divided into normal controls,sham operation,acute lung injury,and EP treatment (40 mg/kg intra-peritoneally every 6 hrs) groups. At the time points of 24 hours the animals in each group were sacrificed, and the lungs were harvested. Wet/dry lung weight ratio, the protein in the bronchoalveolar lavage fluid(BALF),and pulmonary permeability index(PPI) were determined. The histological morphology of lung was observed under microscope. The expression of HMGB1 mRNA was measured using semi-quantitative RT-PCR. Results: EP treatment decreased wet/dry lung weight ratio, the protein in the BALF,and PPI (P<0.01). The histological morphology of lung injury was ameliorated. EP significantly inhibited the HMGB1 mRNA expression(P<0.01). HMGB1 mRNA expression in lungs positively correlation with wet/dry lung weight ratio, the protein in the BALF,and PPI. Conclusion: EP administered inhibits HMGB1 mRNA expression, and protects the lungs against acute injury induced by sepsis.
文摘目的探讨高迁移率蛋白1(HMGB1)在脂多糖诱导的急性肺损伤中的作用,以及甲基强的松龙注射剂干预后其在急性肺损伤中的表达变化。方法将54只清洁级成年雄性SD大鼠随机分成3组:1正常对照组(N组):每只大鼠经尾静脉途径给予生理盐水2 m L/kg,1 h后再经尾静脉途径给予生理盐水1 m L/kg;2急性肺损伤组(ALI组):尾静脉注射脂多糖6 mg/kg,1 h后再经尾静脉途径给予生理盐水1 m L/kg;3甲基强的松龙干预组(MPN组):每只大鼠经尾静脉途径给予脂多糖6 mg/kg,1 h后再经尾静脉途径给予甲基强的松龙20 mg/kg。三组分别于注射后12、24、36 h麻醉处死后采集标本,每个时相点取6只大鼠。采用免疫组织化学法和实时定量PCR(Real-Time PCR)法检测各组各时相点大鼠肺组织HMGB1表达水平,同时观察肺组织病理学改变,并测定干湿重比(W/D)。采用SPSS 20.0统计学软件分析,数据处理采用方差分析。结果 N组中大鼠肺组织有少量HMGB1m RNA及其蛋白表达,ALI组和MPN组大鼠肺组织中HMGB1 m RNA及其蛋白表达水平明显高于N组,差异均有统计学意义(均P<0.05),24 h表达达峰值;同一时相点ALI组大鼠肺组织HMGB1 m RNA及其蛋白表达水平高于MPN组,差异有统计学意义(P<0.05)。ALI组大鼠与MPN组不同时相点W/D增高,与N组比较差异有高度统计学意义(P<0.01),MPN组与ALI组比较W/D下降(P<0.05或P<0.01)。与N组比较,ALI组和MPN组组织病理学有不同程度的病理损害,MPN组比ALI组坏死程度轻,炎症细胞浸润少。结论 HMGB1在炎症后期持续性高水平表达,在ALI发生、发展过程中,尤其是ALI后期失控性炎性反应过程中可能发挥重要作用。甲基强的松龙注射剂通过影响HMGB1 m RNA及其蛋白表达,对脂多糖所致的急性肺损伤有一定的保护作用。
文摘高迁移率族蛋白1(high mobility group box1,HMGB1)是一种高度保守的非组蛋白染色体结合蛋白。在生理条件下,HMGB1参与核内DNA的转录、复制、修复、重组和核小体形成。在病理条件下,大量释放到细胞外的HMGB1作为重要的炎症介质启动和维持炎症反应。HMGB1已被证实与脓毒症、创伤性损伤、自身免疫性疾病以及缺血/再灌注损伤等多种疾病的发病机制密切相关。甘草酸(glycyrrhizic acid)是大量存在于甘草的根和根茎中的一种天然三萜皂苷,具有多种药理功能。研究发现甘草酸作为天然的HMGB1抑制剂,通过与HMGB1结合进而阻断HMGB1与其受体的相互作用从而发挥抗炎活性。本文综述近年来甘草酸及其衍生物治疗HMGB1相关炎症性疾病的药理作用研究,为甘草酸及其衍生物的进一步临床应用提供实验基础。
文摘高迁移率族蛋白B1(high mobility group protein box 1,HMGB1)是一组高度保守的DNA结合蛋白,存在于体内各种细胞的细胞核中,调控着细胞基因的转录。生理状态下HMGB1发挥着核结合蛋白的作用。当释放到细胞间隙后则会起到晚期炎症因子的作用。最近的研究提示,HMGB1不仅参与脓毒症、自身免疫性疾病、慢性肝病、恶性肿瘤等的致病过程,还可参与细胞损伤的修复或致病,在多种疾病及组织器官损伤、修复过程中发挥着重要作用。