草酸钙晶体刺激下人巨噬细胞高迁移率族蛋白B1的表达

背景：研究发现在肾结石模型中肾间质晶体周围存在大量单核/巨噬细胞浸润,显示巨噬细胞可能参与晶体在肾脏中的沉积过程,而巨噬细胞是人体重要的固有免疫细胞,在肾脏中沉积的大量的单核/巨噬细胞吞噬晶体,会产生一些炎症因子损伤和破坏肾小管上皮细胞,最终有利于结石的形成。目的：探讨一水草酸钙晶体刺激人巨噬细胞后高迁移率族蛋白B1的表达水平。方法：用100 mg/L的一水草酸钙刺激巨噬细胞,分别于刺激后0,6,12,24和36 h,用Western blot检测细胞总蛋白和细胞浆内高迁移率族蛋白B1的含量;用实时荧光定量PCR,检测细胞中高迁移率族蛋白B1 mRNA表达情况;分别于一水草酸钙刺激后0,1,2和4 h,用酶联免疫吸附实验测定细胞培养液上清中肿瘤坏死因子α和白细胞介素6的水平。结果与结论：一水草酸钙刺激后0-6 h细胞浆内高迁移率族蛋白B1的含量较低,刺激后12-36 h细胞浆内高迁移率族蛋白B1逐渐增加。一水草酸钙刺激后0-6 h,巨噬细胞总蛋白中高迁移率族蛋白B1含量不高,在刺激后12 h细胞总蛋白高迁移率族蛋白B1的含量开始增加,并且在刺激后24-36 h保持在较高水平。RT-PCR结果显示,一水草酸钙刺激后0-12 h,培养细胞中高迁移率族蛋白B1的mRNA表达量无明显变化,刺激后18-24 h培养细胞中高迁移率族蛋白B1的mRNA表达量明显增加。ELISA结果显示,一水草酸钙刺激后2 h,细胞培养液上清中肿瘤坏死因子α和白细胞介素6表达和释放增加,4 h达到明显高峰。结果表明,一水草酸钙可以诱导人巨噬细胞高迁移率族蛋白B1的表达及mRNA表达增加;诱导人巨噬细胞内肿瘤坏死因子α和白细胞介素6的表达增加,且高迁移率族蛋白B1表达的时间明显晚于肿瘤坏死因子a和白细胞介素6的释放时间。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

冠心病患者高迁移率族蛋白B1水平变化的临床研究

目的:探讨冠心病(CHD)患者血清高迁移率族蛋白B1(HMGB1)水平变化及其临床意义。方法:CHD患者158例,分为急性冠脉综合征组(ACS组,87例)和稳定型心绞痛组(SA组,71例),选择30例年龄和性别匹配的健康体检者作为对照组,采用ELISA检测各组人群血清HMGB1水平,并同时检测超敏C-反应蛋白(hs-CRP)、白细胞计数(WBC)和血脂水平。结果:CHD患者血清HMGB1和hs-CRP水平较对照组明显增高(6.47±1.39ng/ml vs 2.19±0.43ng/ml,P<0.01;3.42±0.87g/ml vs 1.15±0.22g/ml,P<0.01)。ACS组患者血清HMGB1水平显著高于SA组(8.52±2.08ng/ml vs 5.38±1.26ng/ml,P<0.01)。ACS组患者血清hs-CRP水平较SA组和对照组也明显升高。CHD患者血清HMGB1水平与hs-CRP水平呈显著正相关(r=0.685,P<0.01)。结论:血清HMGB1水平的变化在CHD的发病过程中有重要意义,可作为CHD的监测指标之一。     

高迁移率族蛋白B1(HMGB-1)与器官纤维化研究进展

高迁移率族蛋白B1(HMGB1)作为一种重要的晚期炎症因子和促炎因子,可以通过损伤/坏死细胞被动释放,也可由活化状态的细胞主动分泌至胞外介导炎症反应.一直以来,HMGB1在各种急慢性炎症中的作用研究备受关注.器官组织纤维化为持续慢性炎症的后期病理变化,近年来,HMGB1在各种器官纤维化中的作用研究越来越多.许多研究结果显示,HMGB1在肝、肺、肾、心脏等器官纤维化的发生发展中发挥重要作用.     

高迁移率族蛋白B1在神经病理性疼痛中的研究进展

高迁移率族蛋白B1（high mobility group protein B1,HMGB1）是一组高度保守的非组蛋白染色体结合蛋白,几乎存在于所有真核细胞中。在生理状态下,HMGB1发挥着结合细胞核蛋白的作用,一旦释放进入细胞间隙,则表现出炎性因子的作用。一直以来,HMGB1因在各种急慢性炎症中的作用而备受关注,近年的研究表明HMGB1在神经病理性疼痛的发生和发展中也发挥着重要的作用。     

高迁移率族蛋白B1在脂多糖致幼年大鼠脑损伤中的表达

目的 探讨高迁移率族蛋白B1(high mobility gnmp box 1,HMGB1)在脂多糖(LPS)致幼年大鼠脑损伤中的表达变化.方法 SD大鼠随机分为对照组(NS组,n=80),颈外动脉注射生理盐水;脂多糖组(LPS组,n=80),颈外动脉注射LPS,于注射药物后6、12、24、48、72 h处死,甲酰胺法测脑组织伊文思蓝(EB)含量;免疫组织化学技术检测脑组织HMGBI、神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)的表达;RT-PCR技术检测HMGB1 mRNA的表达变化.结果 LPS组脑组织的EB含量、NSE、GFAP蛋白表达均为6 h开始增加,24 h达高峰;HMGB1蛋白及HMGB1 mRNA于6 h开始增加,24 h达高峰,与EB含量、NSE、GFAP蛋白含量呈正相关.各时间点与NS组比较,差异显著.结论 HMGB1可能作为晚期炎症因子参与LPS致脑损伤的发生发展过程.     

高迁移率族蛋白B1在肿瘤中的双重作用

高迁移率族蛋白B1(high mobility group box 1,HMGB1)是广泛存在于真核细胞核染色质中的一种非组蛋白,在机体炎症性疾病、自身免疫病以及癌症中发挥重要作用。在肿瘤的发生发展中,不同亚细胞定位的HMGB1发挥不同的功能。一方面,HMGB1能够促进肿瘤细胞的生长、增殖、侵袭和转移,介导免疫逃逸与免...     

ACI患者血浆高迁移率族蛋白B1的检测及临床意义

目的:检测急性脑梗死(ACI)患者血浆高迁移率族蛋白B1(HMGB1)浓度,分析其与预后的相关性。方法:50例体检健康人静脉血在体检时获得,90例ACI患者静脉血在入院时获得。ELISA测定血浆高迁移率族蛋白B1浓度,统计分析其与预后的相关性。结果:ACI患者血浆HMGB1浓度(8.4±3.8)ng/ml显著高于体检健康者(1.2±0.4)ng/ml(P<0.01),与美国国立卫生院神经功能缺损评分显著正相关(r=0.591,P<0.01),是ACI患者3个月内死亡的独立危险因素(OR=1.892,95%CI=1.145～5.439,P<0.01),可显著预测ACI患者3个月内死亡(曲线下面积=0.842,95%CI=0.772～0.917,P<0.01),其浓度>8.6 ng/ml,对预测ACI患者3个月内死亡有84.2%的灵敏度和77.4%的特异度。结论:ACI后血浆HMGB1浓度升高,与ACI患者不利的临床预后相关。     

高迁移率族蛋白B1:从核蛋白到新的细胞因子

对高迁移率族蛋白B1（HMGB1）的认识曾长期局限于其调节基因转录的核蛋白功能。由于其作为细菌内毒素致死性的晚期介质这一重要功能不久前被发现,HMGB1新的生物学活性及其作用机制引起广泛兴趣和深入研究。HMGB1具有细胞因子的各种共同特性,包括其自分泌和旁分泌的特征、其受体依赖性以及它与其他炎症细胞因子相互诱生与协同作用的网络性。在感染性和非感染性的炎症、损伤和细胞坏死中,HMGB1介导了以巨噬细胞为主的固有免疫应答,发挥了至关重要的前炎症细胞因子和趋化因子的作用,同时还发现其对神经突触和肿瘤细胞的生长的促进作用。以HMGB1为靶子的治疗策略,包括对其产生和释放的抑制以及对其与受体（已知RAGE、TLR2和TLR4）结合的拮抗措施,在实验动物获得了可喜的成功并给临床应用带来了希望。     

血浆高迁移率族蛋白B1浓度与脑外伤预后的相关性分析

目的：揭示脑外伤患者血浆高迁移率族蛋白B1（HMGBl）浓度,分析其与预后的相关性。方法：收集重型脑外伤患者118例和同期健康体检正常者50例。健康体检正常者静脉血体检时获得,脑外伤患者静脉血在入院时获得。ELISA测定血浆HMGBl浓度,统计分析其与预后的相关性。结果：￡检验显示,脑外伤患者血浆HMGBl浓度（9．4±2．3）ng／ml显著高于健康体检正常者（1．6±0．3）ng／ml（t=9．583,P〈0．01）。多因素分析显示,血浆HMGBI浓度是脑外伤1周内死亡（OR=1．309,95％CI=1．197-1．743,P〈0．01）、1年内死亡（OR=1．476,95％CI=1．208-1．796,P〈0．01）和1年内神经功能预后不良（格拉斯哥预后评分1-3分）（OR=1．419,95％CI=1．218-1．697,P〈0．01）的独立危险因素,ROC曲线分析显示,血浆HMGBl浓度可预测脑外伤1周内死亡（曲线下面积=0．865,95％CI=0．824-0．901,P〈0．01）、1年内死亡（曲线下面积=0．891,95％CI=0．835-0．934,P〈0．01）和1年内神经功能预后不良（曲线下面积=0．876,95％CI=0.832-0．925,P〈0．01）。结论：脑外伤后血浆HMGBl浓度显著升高,临床检测这些指标有助于早期判断脑外伤预后。     

"晚期炎症介质"高迁移率族蛋白-1的研究进展

目的 :脓毒症是严重烧 (创 )伤、休克、外科大手术后常见的并发症 ,进一步发展可导致脓毒性休克、多器官功能障碍综合征 ,是当前烧、创伤外科面临的棘手难题 ,已成为临床危重患者的重要死亡原因之一。既往普遍认为 ,“早期”致炎细胞因子[包括肿瘤坏死因子 (TNF -α)、白细胞介素 - 1等 ]是引起机体失控性炎症反应与组织损害的关键介质。新近的研究发现 ,高迁移率族蛋白 - 1(HMGB - 1)可能作为新的“晚期”炎症因子参与了内毒素的致病过程。我们在国家“973”项目和国家杰出青年基金的资助下 ,系统探讨了HMGB - 1在严重烧 (创 )伤后脓…     

Recombinant thrombomodulin ameliorates experimental autoimmune encephalomyelitis by suppressing high mobility group box 1 and inflammatory cytokines

Recombinant thrombomodulin (rTM) has pleiotrophic properties, including anti‐coagulation and anti‐inflammation; however, its effectiveness as a treatment for multiple sclerosis (MS) has not been evaluated fully. High mobility group box 1 (HMGB1) and proinflammatory cytokines, working as inflammatory mediators, are reportedly involved in the inflammatory pathogenesis of MS. The aim of this study was to determine whether rTM can be a potential therapeutic agent for experimental autoimmune encephalomyelitis (EAE). EAE mice received rTM treatment (1 mg or 0·1 mg/kg/day) from days 11 to 15 after immunization. The clinical variables, plasma levels of inflammatory cytokines and HMGB1 and pathological findings in EAE were evaluated. rTM administration ameliorated the clinical and pathological severity of EAE. An immunohistochemical study of the spinal cord showed weaker cytoplasmic HMGB1 staining in the rTM‐treated EAE mice than in the untreated EAE mice. Plasma levels of inflammatory cytokines and HMGB1 were suppressed by rTM treatment. In conclusion, rTM down‐regulated inflammatory mediators in the peripheral circulation and prevented HMGB1 release from nuclei in the central nervous system, suppressing EAE‐related inflammation. rTM could have a novel therapeutic potential for patients with MS.     

高迁移率族蛋白B-1与炎症关系的研究进展

在脓毒症发生早期就采取相应的针对肿瘤坏死因子-α(TNF-α)、白细胞介素-1(IL-l)的抗炎治疗是十分困难的.而且这种治疗并未取得预期的良好治疗效果.     

高迁移率族蛋白B1在风湿性疾病中的作用及机制

高迁移率族蛋白B1(HMGB1)是一种非组蛋白性核蛋白,除了能与DNA结合并调节着染色体的架构外,它还能作为一种损伤相关的分子模式(DAMP)参与风湿性疾病的病理过程.尽管HMGB1主要位于细胞核内,但它在细胞活化和细胞死亡时可以转位至细胞质以及细细胞外空间.在细胞外,HMGB1的活性随半胱氨酸残基的氧化还原状态的不同而发生改变,其中半胱氨酸残基是其与TLR4结合所必需的.HMGB1除了直接刺激细胞外,它还能通过与细胞因子以及其他内源和外源的因子形成免疫复合物而发挥作用.在类风湿性关节炎患者滑液以及该疾病的动物模型中,核外HMGB1的表达量增加.在动物模型中阻断HMGB1表达能使疾病减轻.在系统性红斑狼疮(SLE)中能与DNA相互作用的HMGB1是免疫复合物的组成部分.总之,HMGB1可以是风湿性疾病中的一个重要中介物和生物标志,有希望成为有价值的新靶点用于治疗相关疾病.     

Effects of propofol on early and late cytokines in lipopolysaccharide-induced septic shock in rats

Objective

It has been reported that the intravenous anesthetic propofol (PPF) has anti-inflammatory effects in vitro and in patients. The purpose of this study was to investigate whether PPF has anti-inflammatory effects in lipopolysaccharide (LPS)-induced septic shock by inhibiting the induction of pro-inflammatory cytokines [interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α)] and high mobility group box 1 (HMGB1) in rats.

Methods

Thirty six male Wistar rats were randomly assigned to one of three groups (control group, PPF + LPS group and LPS group; n = 12 per group). Control group rats received a 0.9% NaCl solution (NS) by the tail vein. The PPF + LPS group rats received PPF (10 mg/kg bolus, followed by infusion at 10 mg/(kg·h) through a femoral vein catheter) 1 h before LPS (7.5 mg/kg) administration via the tail vein. The LPS group rats received injection of LPS (7.5 mg/kg) via the tail vein. Hemodynamic effects were recorded as well as mortality rates, and plasma cytokine con-centrations (TNF-α, IL-6, HMGB1) were measured for the 24-h observation period.

Results

The mean arterial pressure and heart rate of the PPF + LPS group were more stable than those of the LPS group. The mortality at 24 h after the administration of the LPS injection was much higher in the LPS group (58.3%) compared to the PPF + LPS group (25.0%). Plasma concentrations of cytokines (IL-6 and TNF-α) and HMGB1 were significantly reduced in the PPF + LPS group compared to the LPS group (P < 0.05).

Conclusion

Pretreatment with PPF reduced the mortality rate of rats and attenuated the pro-inflammatory cytokine responses in an endotoxin shock model through an anti-inflammatory action inhibiting induction of HMGB1.     

The effects of inflammatory cytokines on steroidogenic acute regulatory protein expression in macrophages

Objective: To investigate the expression of steroidogenic acute regulatory protein (StAR) in macrophages and the effects of inflammatory cytokines on StAR expression. Methods: The macrophages isolated from ApoE knockout mice and C57BL/6J mice and RAW264.7 cells (a cell line from mouse macrophage. ATCC Number: TIB-71^TM) were cultured in DMEM containing 10% fetal bovine serum. RAW264.7 cells were treated with different inflammatory cytokines (TNF-α, IFN-γ and TGF-β1) and 8-Br-cAMP, a cAMP analog. RT-PCR and Western blot analysis were applied to evaluate the effects of inflammatory cytokines on StAR expression. Results: RT-PCR and Western blot analysis demonstrated the expression of StAR in the macrophages isolated from ApoE knockout mice, C57BL/6J mice and RAW264.7 cells. Proinflammatory cytokines TNF-α and IFN-γ significantly decreased StAR mRNA and protein levels in RAW264.7 cells. The inhibition was dose- and time-dependent. In contrast, anti-inflammatory cytokine TGF-β1 increased StAR mRNA and protein levels. At 1:15 molecular ratio, TGF-β1 blocked the down-regulation of StAR expression mediated by TNF-α. cAMP also induced StAR expression in RAW264.7 cells. When the cells were co-treated with 8-Br-cAMP and TNF-α, 8-Br-cAMP failed to induce StAR expression. Conclusion: Our results provide interesting evidence that inflammatory cytokines regulate StAR expression in macrophages. Received 12 August 2006; returned for revision 28 September 2006; returned for final revision 28 May 2007; accepted by M. Katori 22 June 2007     

中药骨康对成骨细胞IL-1、IL-6和TNF-α表达的影响

目的 通过观察骨康含药血清对成骨细胞IL-1、IL-6和TNF-α表达的影响，探讨其治疗骨质疏松的作用机制。方法 体外分离、培养成骨细胞，实验分为3组：正常血清组、中药骨康血清组和雌二醇含药血清组。采用酶联免疫吸附法，检测成骨细胞IL-1、IL-6和TNF-α表达。结果 中药骨康含药血清组IL-1、IL-6和TNF-α含量显著低于空白对照组（P〈0．05），中药骨康含药血清组与雌二醇含药血清组IL-1、IL-6和TNF-α的含量无明显差异。结论 在去势状态下，中药骨康可能是抑制IL-1、IL-6和TNF-α分泌，而达到治疗骨质疏松症的作用。     

Regulatory effects of IL-13 on synovial fluid macrophages and blood monocytes from patients with inflammatory arthritis.

Activated macrophages are central to the destructive processes of chronic inflammatory arthritis. In this study, it was hypothesized that IL-13, a product predominantly of 'Th2-type' lymphocytes, may be used therapeutically to down-regulate monocyte/macrophage activities at sites of chronic inflammation. Synovial fluid mononuclear cells were isolated from 12 patients with chronic inflammatory arthritis. Peripheral blood mononuclear cells (PBMC) were isolated at the same time as synovial fluid cells from all 12 patients. IL-13 significantly inhibited lipopolysaccharide (LPS)-induced tumour necrosis factor-alpha (TNF-alpha) production by mononuclear cells from peripheral blood, but not synovial fluid. In contrast, IL-13 inhibited LPS-induced IL-1 beta production by all cells, and as a positive response to IL-13, CD23 expression was increased on both cell populations. Blood monocytes cultured for 7 days with granulocyte-macrophage colony-stimulating factor (GM-CSF) or M-CSF responded to IL-13 in a manner similar to that detected for synovial fluid-derived cells, with suppression of LPS-induced IL-1 beta, but not TNF-alpha, production. In all experiments, the responses to IL-13 were very similar to those detected to IL-4, but differed from those measured with IL-10. Thus, the responses to IL-13 by synovial fluid cells and cultured monocytes are not equal to those of blood monocytes. The similar responses to IL-4 and IL-13 support claims of a common element for signalling from the IL-4 and IL-13 receptors. Furthermore, the activity of a common receptor chain may be altered by monocyte activation and differentiation.     

The novel inflammatory cytokine high mobility group box protein 1 (HMGB1) is expressed by human term placenta

High mobility group box protein 1 (HMGB1) was previously considered a strict nuclear protein, but lately data are accumulating on its extranuclear functions. In addition to its potent proinflammatory capacities, HMGB1 has a prominent role in a number of processes of specific interest for the placenta. Our overall aim was to investigate the expression of HMGB1 in human term placenta and elucidate a potential difference in HMGB1 expression comparing vaginal deliveries with elective Caesarean sections. In addition, placentas from normal pregnancies were compared with placentas from pregnancies complicated by pre-eclampsia. Twenty-five placentas, 12 from normal term pregnancies and 13 from pregnancies complicated by pre-eclampsia were analysed with immunohistochemistry for HMGB1 and its putative receptors; receptor for advanced glycation end-products (RAGE), Toll-like receptor 2 (TLR2) and TLR4. We present the novel finding that in addition to a strong nuclear HMGB1 expression in almost all cells in investigated placentas, an individual variation of cytoplasmic HMGB1 expression was detected in the syncytiotrophoblast covering the peripheral chorionic villi, by cells in the decidua and in amnion. Production of HMGB1 was confirmed by in situ hybridization. Although labour can be described as a controlled inflammatory-like process no differences in HMGB1 expression could be observed comparing active labour and elective Caesarean sections. However, a tendency towards a higher expression of cytoplasmic HMGB1 in the decidua from women with pre-eclampsia was demonstrated. The abundant expression of the receptors RAGE, TLR2 and TLR4 implicates a local capability to respond to HMGB1, although the precise role in the placenta remains to be elucidated.     

高迁移率族蛋白B1基因的克隆及其与神经管发育的相关关系

目的:构建神经管cDNA文库,寻找神经管发育相关基因。方法:提取E8.5 d金黄地鼠神经管总RNA;SMART技术构建神经管cDNA噬菌体表达文库;重组噬菌体PCR鉴定。将出现频率很高的重组噬菌斑,经质粒转化、酶切鉴定和DNA序列分析,证实为高迁移率族蛋白B1基因(HMGB1)。将HMGB1 cDNA片段回收、纯化,制备探针;Northern杂交检测不同发育阶段神经管中HMGB1 mRNA的表达变化。结果:构建的HMGB1cDNA片段含有完整的cDNA序列。Northern杂交显示:随胚胎发育,神经管HMGB1 mRNA表达量逐渐增加,E10 d增加最为明显,E12 d仍处于较高水平;而8.5 d高温致畸胚神经管,HMGB1 mRNA表达量较对照组明显减少。结论:HMGB1基因的表达与神经管发育及高温致神经管畸形的发生密切相关。