High mobility group box 1 protein (HMGB1) as an immune-modulating factor for polarization of human T lymphocytes

Objective This study was performed to investigate the effect of high mobility group box-1 protein (HMGB 1) on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells (PBMCs) were isolated,then rhHMGB 1 was added to PBMCs.Four-color flow cytometric (FCM) analysis was used for the measurement of intracellular cytokine including interleukin Results (1) Different stimulating time and dosage of rhHMGB 1 did not alter the number of IFN-a positive cells (Th 1).rhHMGB 1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th 1 to Th2.(2) Compared with the untreated cells,when the cells were coincubated with rhHMGB 1 (10-100ng/ml) for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGBI.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.     

卡介苗对急性哮喘发作者Th1/Th2及气道炎症的影响及降低住院率作用的研究

目的 探讨卡介苗对急性哮喘发作者 Th1/Th2及气道炎症的影响及降低住院率的作用.方法 选取98例急性哮喘患者,随机分为观察组与对照组,每组49例.对照组采取常规支气管舒张剂解痉平喘治疗,观察组在对照组基础上口服卡介苗.比较2组治疗前后的哮喘症状评分、肺功能指标、IL-4、干扰素γ(IFN-γ)水平、外周血Th1/Th2细胞比、不良反应总发生率及随访1年期间的再住院情况.结果 观察组治疗后的哮喘症状评分为 (1.61±0.36)分,低于对照组 (t=17.961, P<0.05);FEV1%pred与最大呼气流速占预计值百分比 (PEV%pred)分别为 (71.56±9.44)%、(79.41±5.12)%,高于对照组 (t=8.062、13.599,P值均<0.05).观察组治疗后的IL-4为 (21.22± 3.41)ng/L,低于对照组 (t=9.567,P<0.05);IFN-γ为 (154.36±9.67)ng/L,高于对照组 (t=22.041,P<0.05).观察组治疗后的 Th1、Th1/Th2分别为 (17.62±3.77)%、(3.78±0.64),高于对照组 (t=4.634、14.058,P值均<0.05);Th2为 (4.66±1.11)%,低于对照组 (t=10.637,P<0.05).2组不良反应总发生率为12.24%、10.20%,差异无统计学意义 (χ2=0.102,P >0.05).观察组急性加重再住院率为14.28%,低于对照组的33.33% (χ2=4.863,P <0.05);平均发作次数[(1.21±0.08)次]少于对照组 [(1.41±0.12)次](t=9.193,P<0.05).结论 口服卡介苗治疗急性哮喘可有效纠正Th1/Th2失衡、抑制气道炎症反应,缓解患者症状,且可减少发作、降低住院率,操作简便、安全性高.     

人高迁移率族蛋白B1在风湿免疫性疾病中的研究进展

人高迁移率族蛋白B1(HMGB1)是细胞核内典型的非组蛋白。HMGB1在细胞和组织中分布十分广泛,受到炎症刺激后能从坏死或损伤细胞被动释放,并可诱导树突状细胞上调其成熟的表面抗原。本文详细介绍了HMGB1的结构、功能特点及HMGB1在系统性红斑狼疮、类风湿关节炎、肌炎、系统性硬化症、干燥综合征等风湿免疫性疾病中的表达情况。随着对HMGB1研究的不断深入,HMGB1在风湿性疾病诊断、治疗等中的潜力将进一步得到发展。     

冠心病患者高迁移率族蛋白1的表达与意义

目的 探讨冠心病患者危险程度与血清高迁移率族蛋白B1 (HMGB1)的相关性.方法 ELISA法检测冠心病患者及健康对照人群血清HMGB1蛋白水平.结果 急性冠脉综合征(ACS)患者中HMGB1蛋白水平明显高于稳定型心绞痛(SAP)患者(P<0.05)及健康对照人群(P<0.05),SAP患者HMGB1蛋白水平明显高于健康对照人群(P<0.05).结论 血清HMGB1水平可作为评价冠心病危险程度的指标之一.     

血清HMGB1水平评价哮喘严重程度和控制水平的价值

目的观察哮喘患者血清HMGB1水平与患者严重程度及控制水平的关系。方法检测49例初诊哮喘患者(间歇/轻度20例、中度15例、重度14例)血清HMGB1的水平,并与36例完全控制哮喘组患者及20例正常人对照。结果间歇发作/轻度持续组,中度持续组与重度持续组哮喘患者血清HMGB1水平的差异有统计学意义(P值均<0.05)。完全控制哮喘组血清HMGB1水平显著低于初诊哮喘患者,但显著高于正常对照组(P值均<0.01)。结论血清HMGB1水平可以反应哮喘的严重程度和指导哮喘治疗。     

IRAK-M knockout promotes allergic airway inflammation,but not airway hyperresponsiveness,in house dust mite-induced experimental asthma model

BackgroundIL-1 receptor associated-kinase (IRAK)-M, expressed by airway epithelium and macrophages, was shown to regulate acute and chronic airway inflammation exhibiting a biphasic response in an OVA-based animal model. House dust mite (HDM) is a common real-life aeroallergen highly relevant to asthma pathogenesis. The role of IRAK-M in HDM-induced asthma remains unknown. This study was aimed to investigate the effect of IRAK-M on allergic airway inflammation induced by HDM using IRAK-M knockout (KO) mice and the potential underlying mechanisms.MethodsIRAK-M KO and wild-type (WT) mice were sensitized and challenged with HDM. The differences in airway inflammation were evaluated 24 hours after the last challenge between the two genotypes of mice using a number of cellular and molecular biological techniques. In vitro mechanistic investigation was also involved.ResultsLung expression of IRAK-M was significantly upregulated by HDM in the WT mice. Compared with the WT controls, HDM-treated IRAK-M KO mice showed exacerbated infiltration of inflammatory cells, particularly Th2 cells, in the airways and mucus overproduction, higher epithelial mediators IL-25, IL-33 and TSLP and Th2 cytokines in bronchoalveolar lavage (BAL) fluid. Lung IRAK-M KO macrophages expressed higher percentage of costimulatory molecules OX40L and CD 80 and exhibited enhanced antigen uptake. However, IRAK-M KO didn’t impact the airway hyperreactivity (AHR) indirectly induced by HDM.ConclusionsThe findings indicate that IRAK-M protects allergic airway inflammation, not AHR, by modifying activation and antigen uptake of lung macrophages following HDM stimulation. Optimal regulation of IRAK-M might indicate an intriguing therapeutic avenue for allergic airway inflammation.     

高迁移率族蛋白1及其受体对大鼠骨髓间充质干细胞增殖及细胞因子水平的影响

目的探讨高迁移率族蛋白1(HMGB1)及其受体对大鼠骨髓间充质干细胞(MSC)增殖及细胞因子水平的影响。方法体外传代培养大鼠MSC,以25.0、50.0及100.0μg/L HMGB1分别作用细胞24 h、48 h、72 h后,MTT检测MSC的增殖情况。25.0、50.0及100.0μg/L HMGB1作用MSC 48 h后,ELISA测定细胞培养液中血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(b FGF)水平,Western blot检测MSC中HMGB1的晚期糖基化终产物受体(RAGE)和Toll样受体4(TLR4)的表达情况。确定HMGB1对MSC作用的最佳药物浓度后,siRNA分别抑制RAGE受体和TLR4受体,观察HMGB1对MSC增殖及分泌细胞因子水平的影响。结果 25.0μg/L HMGB1作用MSC 24 h、48 h、72 h后均能有效促细胞增殖(P0.05);25.0、50.0μg/L HMGB1作用细胞48 h后,细胞释放VEGF、b FGF明显增多,RAGE及TLR4表达也增高(P0.05)。特异siRNA转染靶向干扰MSC中TLR4的表达后,促细胞增殖及分泌VEGF、b FGF作用受到抑制(P0.05),而siRNA干扰RAGE表达对HMGB1的细胞效应无明显影响。结论 HMGB1可通过结合其受体TLR4促进MSC增殖以及VEGF、b FGF分泌。     

Th1/Th2漂移与支气管哮喘的免疫治疗

Th1/Th2细胞在不同的细胞因子、抗原等因素的影响下，可发生Th1/Th2的转换，研究表明在支气管哮喘（哮喘）的发生机制中，Th2细胞分化明显增多，而Th1细胞数目减少。因此，促进Th1反应，抑制Th2反应的免疫治疗是治疗哮喘的重要部分。目前许多调节Th1/Th2失衡的免疫治疗方法已取得一些进展。     

类风湿关节炎患者外周血高迁移率族蛋白1表达

目的通过检测类风湿关节炎（RA）患者外周血单个核细胞（PBMC）、血浆中高迁移率族蛋白1（HMGB1）表达,为寻找治疗RA的新靶点提供依据。方法采集38例活动期RA患者、24例相对稳定期RA患者和20例健康对照者外周血。RT．PCR检测PBMC HMGB1mRNA表达,Western blot检测PBMC、血浆HMGB1蛋白表达。结果与相对稳定期RA患者、健康对照者相比,活动期RA患者PBMC HMGB1mRNA表达水平差异无统计学意义（F=1．23,P〉0．05）,而HMGB1蛋白表达水平下降（F=70．91,P〈0．01）,血浆HMGB1水平显著增高（P〈0．001）。相对稳定期RA患者与健康对照者之间差异无统计学意义（P〉0．05）。活动期RA患者血浆HMGB1水平与ESR（r．=0．478。P〈0．001）、C-反应蛋白（rs=0．574,P〈0．05）呈正相关。结论HMGB1与RA发病密切相关,并可能成为新的治疗靶点。     

Delayed ethyl pyruvate therapy attenuates experimental severe acute pancreatitis via reduced serum high mobility group box 1 levels in rats

AIM： To investigate the effect of delayed ethyl pyruvate （EP） delivery on distant organ injury, survival time and serum high mobility group box 1 （HMGB1） levels in rats with experimental severe acute pancreatitis （SAP）.
METHODS： A SAP model was induced by retrograde injection of artificial bile into the pancreatic ducts of rats. Animals were divided randomly into three groups （n = 32 in each group）： sham group, SAP group and delayed EP treatment group. The rats in the delayed EP treatment group received EP （30 mg/kg） at 12 h, 18 h and 30 h after induction of SAP. Animals were sacrificed, and samples were obtained at 24 h and 48 h after induction of SAP. Serum HMGB1, aspartate arninotransferase （AST）, alanine arninotransferase （ALT）, blood urea nitrogen （BUN）, and creatinine （Cr） levels were measured. Lung wet-to-dry-weight （W/D） ratios and histological scores were calculated to evaluate lung injury. Additional experiments were performed between SAP and delayed EP treatment groups to study the influence of EP on survival times of SAP rats.
RESULTS： Delayed EP treatment significantly reduced serum HMGB1 levels, and protected against liver, renal and lung injury with reduced lung W/D ratios （8.22 ±0.42 vs 9.76 ± 0.45, P 〈 0.01）, pulmonary histological scores （7.1 ± 0.7 vs 8.4 ± 1.1, P 〈 0.01）, serum AST （667 ± 103 vs 1 368 ± 271, P 〈 0.01）, ALT （446 ± 91 vs 653 ± 98, P 〈 0.01） and Cr （1.2 ± 0.3 vs 1.8 ± 0.3, P 〈 0.01） levels. SAP rats had a median survival time of 44 h. Delayed EP treatment significantly prolonged median survival time to 72 h （P 〈 0.01）.
CONCLUSION： Delayed EP therapy protects against distant organ injury and prolongs survival time via reduced serum HMGBllevels in rats with experimental SAP. EP may potentially serve as an effective new therapeutic option against the inflammatory response and multiple organ dysfunction syndrome （MODS） in SAP patients.     

Delayed ethyl pyruvate therapy attenuates experimental severe acute pancreatitis via reduced serum high mobility group box 1 levels in rats

AIM: To investigate the effect of delayed ethyl pyruvate (EP) delivery on distant organ injury,survival time and serum high mobility group box 1 (HMGB1) levels in rats with experimental severe acute pancreatitis (SAP).METHODS: A SAP model was induced by retrograde injection of artificial bile into the pancreatic ducts of rats. Animals were divided randomly into three groups (n=32in each group): sham group, SAP group and delayed EP treatment group. The rats in the delayed 18 h and 30 h after induction of SAP. Animals were sacrificed, and samples were obtained at 24 h and 48 h after induction of SAR Serum HMGB1, aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), and creatinine (Cr) levels were measured. Lung wet-to-dry-weight (W/D) ratios and histological scores were calculated to evaluate lung injury. Additional experiments were performed between SAP and delayed EP treatment groups to study the influence of EP on survival times of SAP rats.RESULTS: Delayed EP treatment significantly reduced serum HMGB1 levels, and protected against liver, renal and lung injury with reduced lung W/D ratios (8.22±0.42vs 9.76±0.45, P < 0.01), pulmonary histological scores (7.1±0.7 vs 8.4±1.1, P < 0.01), serum AST (667± 103 vs 1 368 ± 271, P < 0.01), ALT (446±91vs 653±98, P < 0.01) and Cr (1.2 ± 0.3 vs 1.8±0.3,P < 0.01) levels. SAP rats had a median survival time of 44 h. Delayed EP treatment significantly prolonged median survival time to 72 h (P < 0.01).CONCLUSION: Delayed EP therapy protects against distant organ injury and prolongs survival time via reduced serum HMGBllevels in rats with experimental SAP. EP may potentially serve as an effective new therapeutic option against the inflammatory response and multiple organ dysfunction syndrome (NODS) in SAP patients.     

支气管哮喘急性发作期Th1/Th2细胞失衡与峰流速变化相关性的临床研究

目的研究支气管哮喘（哮喘）急性发作期患者Th1／Th2细胞失衡与最大呼气流速（PEF）昼夜变化的相关性。方法哮喘组40例，其中男性24例，女性16例，平均年龄（45．6±10．9）岁，依严重程度分为A组（轻度组）、B组（中重度组）。测定支气管舒张试验，并用电子峰流速仪每6h检测一次PEF的变化，同时检测外周静脉血Th1和Th2细胞。健康对照组35例，其中男性18例，女性17例，平均年龄（46．2±11．8）岁，为门诊查体健康者，均无吸烟史及过敏史。结果与健康对照组相比，哮喘A组、B组Th2细胞增高，Th1／Th2比值降低，Th1细胞无显著性统计学差异（P=0．079）；哮喘B组与A组比较，Th2细胞增高。Th2细胞与PEF日内变异率呈正相关，与4AMPEF值呈负相关。结论哮喘急性发作期患者存在Th1／Th2细胞失衡，以Th2细胞增高为主，中重度患者Th2细胞增高更明显。哮喘急性发作期患者Th2细胞增高与PEF日内变异率有关，且主要是与PEF谷值降低有关，Th2细胞可能在PEF变异率中发挥重要作用。     

High-mobility group box 1 protein and its role in severe acute pancreatitis

The high mobility group box 1(HMGB1),which belongs to the subfamily of HMG-1/-2,is a highly conserved single peptide chain consisting of 215 amino acid residues with a molecular weight of approximately 24894 Da.HMGB1 is a ubiquitous nuclear protein in mammals and plays a vital role in inflammatory diseases.Acute pancreatitis is one of the most common causes of acute abdominal pain with a poor prognosis.Acute pancreatitis is an acute inflammatory process of the pancreas(duration of less than six months),for which the severe form is called severe acute pancreatitis(SAP).More and more studies have shown that HMGB1 has a bidirectional effect in the pathogenesis of SAP.Extracellular HMGB1 can aggravate the pancreatic inflammatory process,whereas intracellular HMGB1 has a protective effect against pancreatitis.The mechanism of HMGB1 is multiple,mainly through the nuclear factor-κB pathway.Receptors for advanced glycation endproducts and toll-like receptors(TLR),especially TLR-2 and TLR-4,are two major types of receptors mediating the inflammatory process triggered by HMGB1 and may be also the main mediators in the pathogenesis of SAP.HMGB1 inhibitors,such as ethyl pyruvate,pyrrolidine dithiocarbamate and Scolopendra subspinipes mutilans,can decrease the level of extracellular HMGB1 and are the promising targets in the treatment of SAP.     

心房颤动患者高迁移率族蛋白1血清水平

目的探讨心房颤动(AF)患者血清高迁移率族蛋白1(HMGB1)水平是否升高。方法入选的55例阵发性AF患者,33例持续性AF患者,选择36例年龄和性别匹配的无AF体检者作为对照组,采用ELISA法检测血清HMGB1和高敏C反应蛋白(hs-CRP)水平。结果持续性AF患者血清HMGB1水平较对照组和阵发性AF患者显著增高(8.62±2.18 ng/m l vs 2.22±0.63 ng/m l,5.29±1.43 ng/m l,P均<0.05)。阵发性AF患者血清HMGB1水平较对照组高(P<0.05)。AF患者血清HMGB1水平与hs-CRP水平呈显著正相关(n=88,r=0.629,P<0.05)。结论 AF患者血清HMGB1水平显著上升。     

Calycosin attenuates severe acute pancreatitis-associated acute lung injury by curtailing high mobility group box 1 - induced inflammation

BACKGROUNDAcute lung injury (ALI) is a common and life-threatening complication of severe acute pancreatitis (SAP). There are currently limited effective treatment options for SAP and associated ALI. Calycosin (Cal), a bioactive constituent extracted from the medicinal herb Radix Astragali exhibits potent anti-inflammatory properties, but its effect on SAP and associated ALI has yet to be determined. AIMTo identify the roles of Cal in SAP-ALI and the underlying mechanism.METHODSSAP was induced via two intraperitoneal injections of L-arg (4 g/kg) and Cal (25 or 50 mg/kg) were injected 1 h prior to the first L-arg challenge. Mice were sacrificed 72 h after the induction of SAP and associated ALI was examined histologically and biochemically. An in vitro model of lipopolysaccharide (LPS)-induced ALI was established using A549 cells. Immunofluorescence analysis and western blot were evaluated in cells. Molecular docking analyses were conducted to examine the interaction of Cal with HMGB1.RESULTSCal treatment substantially reduced the serum amylase levels and alleviated histopathological injury associated with SAP and ALI. Neutrophil infiltration and lung tissue levels of neutrophil mediator myeloperoxidase were reduced in line with protective effects of Cal against ALI in SAP. Cal treatment also attenuated the serum levels and mRNA expression of pro-inflammatory cytokines tumor necrosis factor-α, interleukin-6, IL-1β, HMGB1 and chemokine (CXC motif) ligand 1 in lung tissue. Immunofluorescence and western blot analyses showed that Cal treatment markedly suppressed the expression of HMGB1 and phosphorylated nuclear factor-kappa B (NF-κB) p65 in lung tissues and an in vitro model of LPS-induced ALI in A549 cells suggesting a role for HGMB1 in the pathogenesis of ALI. Furthermore, molecular docking analysis provided evidence for the direct interaction of Cal with HGMB1.CONCLUSIONCal protects mice against L-arg-induced SAP and associated ALI by attenuating local and systemic neutrophil infiltration and inflammatory response via inhibition of HGMB1 and the NF-κB signaling pathway.     

High mobility group 1 B-box mediates activation of human endothelium

OBJECTIVES: Severe sepsis and septic shock is a consequence of a generalized inflammatory systemic response because of an invasive infection that may result in acute organ dysfunction. Mortality is high despite access to modern intensive care units. The nuclear DNA binding protein high mobility group 1 (HMGB1) protein has recently been suggested to act as a late mediator of septic shock via its function as a macrophage-derived pro-inflammatory cytokine (J Exp Med 2000; 192: 565, Science1999; 285: 248). We investigated the pro-inflammatory activities of the A-box and the B-box of HMGB1 on human umbilical venular endothelial cells (HUVEC). DESIGN: The HUVEC obtained from healthy donors were used for experiments. Recombinant human full-length HMGB1, A-box and B-box were cloned by polymerase chain reaction (PCR) amplification from a human brain quick-clone cDNA. The activation of HUVEC was studied regarding (i) upregulation of adhesion molecules, (ii) the release of cytokines and chemokines, (iii) the adhesion of neutrophils to HUVEC, (iv) the activation of signalling transduction pathways and (v) the involvement of the receptor for advanced glycation end-products (RAGE). RESULTS: The full-length protein and the B-box of HMGB1 dose-dependently activate HUVEC to upregulate adhesion molecules such as ICAM-1, VCAM-1 and E-selectin and to release IL-8 and G-CSF. The activation of HUVEC could be inhibited to 50% by antibodies directed towards the RAGE. HMGB1-mediated HUVEC stimulation resulted in phosphorylation of the ELK-1 signal transduction protein and a nuclear translocation of p65 plus c-Rel, suggesting that HMGB1 signalling is regulated in endothelial cells through NF-kappaB. CONCLUSIONS: The HMGB1 acts as a potent pro-inflammatory cytokine on HUVEC and the activity is mainly mediated through the B-box of the protein. HMGB1 may be a key factor mediating part of the pro-inflammatory response occurring in septic shock and severe inflammation.     

高迁移率族蛋白-1在小鼠溃疡性结肠炎发病中的作用研究

目的探讨高迁移率族蛋白-1（HMGBl）在溃疡性结肠炎（UC）发病机制中的作用。方法BALB／c小鼠32只,随机分成2组,即UC模型组（H=24）和正常对照组（n=8）。以3％葡聚糖硫酸钠（DSS）制备小鼠UC模型,造模第24h、96h、168h,分别处死造模组（各8只）;对照组正常饮水。观察其病理变化;ELISA法检测血清HMGBl水平;Western—blot法检测结肠组织HMGBl蛋白的表达。结果随着造模时间的延长,UC组织学评分上升,造模168h时其结肠黏膜表现与人类UC相类似。UC组血清HMGBl水平在造模24h时比正常组略升高[（4．49±0．53）μg／L比（5．09±0．61）μg/L,P〉0．05],在96h达高峰[（14．74±0．60）μg／L,P〈0．01],168h时峰值下降[（9．03±0．78）μg／L,P〈0．01]。UC组小鼠结肠组织HMGBl蛋白表达水平在造模24h时较正常略升高（0．49±0．03比0．56±0．02,P〉0．05）,在造模96h明显升高（0．76±0．03,P〈0．05）,至168h时仍维持在较高水平（0．77±0．04,P〈0．05）。结论HMGB1在UC小鼠血清中水平升高,结肠组织中也表达明显增强,提示HMGB1作为晚期炎症因子可能参与UC的疾病过程。     

日本血吸虫高速泳动家族B1蛋白基因的克隆 表达与功能分析

目的 目的 克隆、 表达日本血吸虫高速泳动家族B1蛋白 （SjHMGB1）, 并研究其功能特性。方法 方法 利用逆转录PCR （RT?PCR） 技术从日本血吸虫成虫mRNA中反转录扩增出编码SjHMGB1的完整开放阅读框DNA片段, 然后将其亚克隆至 原核表达载体质粒pET28a （+） 中, 构建重组表达质粒SjHMGB1?pET28a。将重组表达质粒转化大肠埃希菌BL21 （DE3）, 含 重组质粒的转化子细菌经异丙基硫代半乳糖苷 （IPTG） 诱导以表达重组SjHMGB1蛋白。通过镍螯合琼脂糖亲和胶亲和层 析法制备纯化的重组SjHMGB1蛋白。通过DNA迟滞实验和动物免疫鉴定重组SjHMGB1的DNA结合和免疫学特性。通 过免疫印迹试验 （Western blotting） 和RT?PCR确定SjHMGB1分子在日本血吸虫不同发育阶段的表达情况。以重组SjH? MGB1为抗原免疫小鼠, 3次免疫后每只小鼠感染45±2条血吸虫尾蚴, 42 d后剖杀, 计算减虫率与减卵率, 观察其诱导抗血 吸虫感染的免疫保护性作用。 结果 结果 采用RT?PCR法从日本血吸虫成虫mRNA中反转录扩增到长度约为530 bp的特异性 DNA条带, 序列分析证实为编码SjHMGB1蛋白的开放阅读框序列。将此片段亚克隆至表达质粒pET28a （+） 中, 成功构建 了重组表达质粒SjHMGB1?pET28a。含重组表达质粒SjHMGB1?pET28a的转化子细菌经IPTG诱导能表达分子量约28 kDa 的可溶性SjHMGB1蛋白。采用镍螯合亲和层法制备了纯化的重组SjHMGB1蛋白。DNA迟滞试验显示纯化的Sj HMGB1 蛋白可同超螺旋及线性DNA结合, 重组SjHMGB1免疫小鼠能产生高效价的抗体IgG, Western blotting分析证实重组SjH? MGB1蛋白能被重感染小鼠血清所识别, 表明重组表达的SjHMGB1分子具有天然蛋白的功能活性与免疫学特性。RT?PCR 和Western blotting分析显示SjHMGB1分子在血吸虫成虫和虫卵期大量表达, 而在尾蚴阶段表达量极低。免疫保护性试验 结果表明, 重组SjHMGB1分子免疫不能诱导有效的抗血吸虫感染免疫保护性作用。结论 结论 获得了编码SjHMGB1基因和 纯化的具有天然功能活性的重组SjHMGB1蛋白; 重组SjHMGB1蛋白具有强烈的免疫原性, 但不能诱导有效的抗血吸虫感 染免疫保护性作用。     

丙酮酸乙酯对实验性结肠炎大鼠结肠组织HMGB1表达及细胞因子的影响

目的:观察丙酮酸乙酯(EP)对实验性结肠炎大鼠结肠组织HMGB1表达及炎症因子的影响.方法:SD大鼠随机分为空白对照组、模型组、EP治疗组,每组12只;模型组、治疗组大鼠用DSS溶液复制实验性结肠炎大鼠模型,观察各组实验大鼠DAI评分、大体形态学及组织病理学评分(HPS)改变,采用应用逆转录-聚合酶链反应(RT-PCR)和免疫印迹法(Westernblot)检测结肠组织HMGB1表达;酶联免疫吸附法检测细胞因子(TNF-α、IL-6)的含量.结果:DAI和HPS在DSS模型组中高于空白对照组(7.20±2.28vs0.45±0.16,13.60±0.72vs6.4±0.85,P<0.01),在空白对照组结肠组织中HMGB1低表达,但在模型组表达显著上调(P<0.01);模型组血清TNF-α、IL-6水平高于空白对照组(190.40±24.55vs43.65±8.79,238.75±26.58vs74.3±7.92,P<0.01);与DSS模型组相比,EP可以缓解大鼠体质量的减轻,减少腹泻及便血的发生,并且能够显著下调结肠组织DAI、HPS、HMGB1mRNA和蛋白的表达,下调血清TNF-α、IL-6水平(P<0.05).结论:HMGB1参与了实验性结肠炎大鼠的发病过程,应用EP治疗可有效抑制实验性结肠炎大鼠结肠组织HMGB1的表达,可以明显下调TNF-α、IL-6等促炎因子的水平,有助于减轻实验性结肠炎大鼠炎症反应.