THH对HIF-1α在CIA动物模型中表达的影响及意义

目的:探讨昆明山海棠(THH)对低氧诱导因子(HIF-1α)在胶原关节炎(CIA)大鼠模型中表达的影响及其作用机制的研究。方法:建立CIA大鼠模型,然后用药治疗30 d,30 d后HE染色观察关节的病理改变,RT-PCR以及免疫组化法检测HIF-1α的表达。结果:给药30 d后,HE染色显示治疗组明显减轻滑膜增殖以及炎症反应,免疫组化以及PT-PCR显示治疗组HIF-1α明显表达降低。结论:THH治疗CIA大鼠模型具有显著疗效的作用机制与调节外周血清以及滑膜组织内的HIF-1α的表达有关。     

昆明山海棠对CIA大鼠模型中HIF-1α表达的影响

目的:研究昆明山海棠(THH)对胶原诱导性关节炎(CIA)大鼠模型中低氧诱导因子1α(HIF-1α)表达的影响及其作用机制.方法:建立CIA大鼠模型,随机分组,治疗组分别采用高、中、低剂量THH每天灌胃1次,连续用药30d.动态观察关节炎指数(AI)及关节的病理改变,用RT-PCR及免疫组化染色法分别检测HIF-1 αmRNA及其蛋白的表达.结果:THH可明显抑制CIA大鼠的足爪肿胀,明显减轻滑膜增殖及炎症反应,HIF-1α表达明显降低.结论:THH通过降低HIF-1α的表达,减轻CIA模型大鼠关节的炎症反应.     

昆明山海棠对CIA大鼠模型中HIF-1α表达的影响

目的:研究昆明山海棠(THH)对胶原诱导性关节炎(CIA)大鼠模型中低氧诱导因子1α(HIF-1α)表达的影响及其作用机制。方法:建立CIA大鼠模型,随机分组,治疗组分别采用高、中、低剂量THH每天灌胃1次,连续用药30d。动态观察关节炎指数(AI)及关节的病理改变,用RT-PCR及免疫组化染色法分别检测HIF-1αmRNA及其蛋白的表达。结果:THH可明显抑制CIA大鼠的足爪肿胀,明显减轻滑膜增殖及炎症反应,HIF-1α表达明显降低。结论:THH通过降低HIF-1α的表达,减轻CIA模型大鼠关节的炎症反应。     

缺氧诱导因子-1α在高原低氧脑损伤中的研究进展

缺氧诱导因子-1(HIF-1)是一种调节人体对缺氧反应的关键因子.HIF-1α是HIF-1的活性亚基,为细胞缺氧信号转导通路中的核心因子.缺氧条件下HIF-1α显著增高,调控细胞增殖、凋亡、红细胞及血管生成等重要过程.低压、低氧是高原环境的主要特点,近年来越来越多的平原居民前往高海拔地区工作、旅游,急进高原暴露有发展成...     

THH在CIA大鼠模型中对HIF-1α的影响及其作用机制

目的:研究昆明山海棠(THH)在胶原性关节炎(CIA)大鼠模型中对于低氧诱导因子(HIF-1α)、血管内皮生长因子(VEGF)表达的影响,从而探讨其作用机制。方法:建立Wistar大鼠Ⅱ型胶原诱导性关节炎(CIA)模型,动态观察关节炎指数(AI),HE染色观察关节的病理改变,RT-PCR以及免疫组化法检测HIF-1α的表达。结果:给药30天后,治疗组明显抑制胶原性关节炎大鼠的足爪肿胀,降低关节炎指数,HE染色显示明显减轻滑膜增殖以及炎症反应,免疫组化以及PT-PCR显示治疗组HIF-1α明显表达降低。结论:THH治疗大鼠CIA模型具有显著疗效,研究提示THH通过HIF-1α的途径调节其下游基因的表达可能是治疗RA患者的作用机制之一。     

低氧诱导因子-1α参与低氧预处理诱导的心肌血管生成

目的：研究低氧诱导因子-1α（HIF-1α）在低氧预处理（HPC）诱导心肌血管生成中的作用,并探讨其细胞信号转导的机制。方法：雄性SD大鼠随机分为对照组和HPC组。动物置于低氧仓内，持续通入10% O2和90% N2 4 h复制低氧预处理模型。用Ⅷ因子免疫组化染色检测HPC后7 d和21 d心肌组织微血管密度。制备心肌组织蛋白提取物，分别以磷酸化的细胞外信号调节激酶（ERK1/2）抗体和HIF-1α抗体检测HPC后1 d、7 d和21 d p-ERK1/2活性及HIF-1α表达。结果：HPC后7 d和21 d心肌组织微血管密度分别较对照组高36.99%和37.76%（均P<0.01）；p-ERK1/2活性在HPC后1d 较对照组高18.67%；HPC诱导HIF-1α的表达，其表达高峰出现在HPC后1 d。结论：HPC可以促进心肌微血管生成，其机制涉及ERKs活化和HIF-1α表达上调。     

慢性低氧时小鼠肺组织低氧诱导因子-1α的表达

目的：研究低氧时小鼠肺组织中低氧诱导因子-1α(HIF-k)表达的变化。方法：实验用雄性小鼠,低氧仓浓度分别为10％、7％、5％。用免疫荧光组织化学技术及共聚焦显微术,检测小鼠在低氧条件下肺组织中HIF-1α表达的变化。结果：正常组小鼠肺组织HIF-1α无表达,低氧组HIF-1α表达增加,且随低氧时间的延长及低氧强度的增加而增强。结论：低氧可诱导小鼠肺组织中HIF-1α的表达增强,(HIF-k)可能参与肺组织细胞凋亡的发生。     

HIF-1α和HIF-2α在胃癌中的表达及意义

目的 探讨胃癌中低氧诱导因子HIF-1α和HIF-2α的表达及其临床意义。方法 用免疫组化SP法检测组织中HIF-1α、HIF-2α及VEGF的表达;用Western blot法检测组织中HIF-1α、HIF-2α的表达。结果 胃癌中HIF-1α、HIF-2α和VEGF的阳性表达率分别为61.5%、36.5%和61.5%,均显著高于正常对照组的11.1％、0和0;HIF-1α表达与VEGF及胃癌的淋巴结转移密切相关。胃癌组织中HIF-1α和HIF-2α蛋白的表达显著高于癌旁正常胃黏膜组织。结论 低氧诱导因子的表达可能在胃癌的发生、发展中具有重要作用。     

低氧诱导因子-1α在糖尿病肾病中作用机制的研究进展

低氧诱导因子-1α (HIF-1α)是一种核转录因子。在高糖低氧条件下,HIF-1α表达增加,诱导其下游靶基因VEGF、HO-1和BNIP3的表达,通过影响血管生成、细胞外基质沉积、铁代谢及线粒体自噬参与糖尿病肾病(DKD)的发生发展。此外,HIF-1α通过促进细胞因子的产生,参与DKD炎性反应并导致肾脏纤维化。     

低氧诱导因子-1在类风湿关节炎中的作用

低氧诱导因子-1（HIF-1）是普遍存在于人和哺乳动物细胞中的低氧应答词控因子，通过调控一系列与缺氧适应有关基因的表达以维持机体氧稳态。在低氧条件下，HIF—1表达上调并与靶基因结合，促进靶基因转录，引起一系列细胞对低氧的反应。在类风湿关节炎（RA）中，HIF-1表达增加，并通过促进滑膜血管生成、抑制细胞凋亡、调节糖酵解等途径调控RA的发生发展。基于抗HIF-1的靶向治疗可为RA开辟新的治疗途径。     

The inhibition of Src kinase suppresses the production of matrix metalloproteinases in from synovial fibroblasts and inhibits MAPK and STATs pathways

Background/aimThe purpose of this study was to investigate the antiarthritic potentials of the inhibition of Src kinase in vivo and in vitro settings. Materials and methodsArthritis was induced by intradermal injection of chicken type II collagen combined with incomplete Freund’s adjuvant (collagen induced arthritis [CIA] model) in Wistar albino rats. One day after the onset of arthritis, dasatinib, a potent Src kinase inhibitor, (5 mg/kg/day) was given via oral gavage. Tissue Src, Fyn, MAPK and STAT mRNA expressions were determined by real-time polymerase chain reaction. On the other hand, fibroblast like synoviocytes (FLSs) were harvested patients with rheumatoid arthritis (RA) undergoing surgical knee joint replacement. FLSs were stimulated with cytokines and dasatinib was added in different concentrations. MMP –1, –3, and –13 levels in FLSs culture were determined by ELISA.ResultsThe tissue mRNA expressions of Src, Fyn, MAPK and STATs were increased in the arthritis CIA group compared to the control group. Their mRNA expressions in the CIA + dasatinib group were decreased and similar in the control group. In in vitro setting, MMP –1, –3, and –13 expressions from FLSs induced by IL-1β and TNF-α were increased, while dasatinib suppressed their productions from FLSs.ConclusionThe present study shows that the inhibition of Src kinase has antiarthritic potentials in both in vivo and in vitro settings. Src kinase inhibition may be candidate to further research in human RA.     

低氧诱导的小窝蛋白-1上调参与人肺腺癌细胞A549迁移和侵袭

 目的:探讨低氧诱导剂二氯化钴（CoCl2）对小窝蛋白-1（Cav-1）生成的调节作用及后者对人肺腺癌A549细胞迁移、侵袭的影响。方法:检测肺癌患者伴恶性胸水（MPE）和结核性胸膜炎患者胸水（TBPE）中Cav-1和缺氧诱导因子(HIF)-1α浓度,比较两者相关性;以CoCl2和（或）HIF-1α抑制剂YC-1作用于A549细胞ELISA法检测细胞上清Cav-1和HIF-1α浓度;分别采用细胞划痕实验及Transwell小室侵袭实验研究CoCl2刺激表达的Cav-1对A549细胞迁移和侵袭的影响。结果:MPE中Cav-1和HIF-1α浓度明显高于TBPE,两组患者胸水中Cav-1与HIF-1α均呈正相关。CoCl2浓度和时间依赖性诱导A549细胞Cav-1和HIF-1α生成,200 μmol/L或24 h达到峰值;浓度>200 μmol/L或作用时间超过24 h则呈现浓度或时间依赖性抑制。HIF-1α抑制剂YC-1浓度依赖性抑制HIF-1α和Cav-1生成。CoCl2浓度依赖性增强A549细胞迁移和侵袭,200 μmol/L作用最强;YC-1对上述过程产生抑制效应。结论: 肺癌患者胸腔积液中Cav-1浓度升高,低氧诱导Cav-1生成的变化可能参与了A549细胞迁移和侵袭,HIF-1α可能对Cav-1生成发挥影响。     

Inhibition of histone deacetylase down-regulates the expression of hypoxia-induced vascular endothelial growth factor by rheumatoid synovial fibroblasts

Objective: To investigate the effect of FK228 on the in vitro expression of hypoxia-inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) by rheumatoid arthritis synovial fibroblasts (RASFs), and on the in vivo expression of VEGF and angiogenesis in the synovial tissue of mice with collagen-antibody-induced arthritis (CAIA). Methods: RASFs were stimulated with IL-1β and TNFα and then incubated under hypoxia (1 % O₂) with various concentrations of FK228. The effects of FK228 on the expression of HIF-1α and VEGF mRNA were examined by quantitative real-time PCR. Changes in HIF-1α protein expression and the secretion of VEGF protein into the culture medium were examined by Western blot analysis and ELISA, respectively. Immunohistochemical analysis was carried out to investigate the expression and distribution of VEGF in synovial tissues of CAIA mice. Results: The cytokine-stimulated expression of HIF-1α and VEGF mRNA was inhibited by FK228 in a dose-dependent manner. FK228 also reduced the expression of HIF-1α and VEGF protein. Intravenous administration of FK228 (2.5 mg/kg) suppressed VEGF expression, and also blocked angiogenesis in the synovial tissue of CAIA. Conclusion: FK228 may exhibit a therapeutic effect on RA by inhibition of angiogenesis through down-regulation of angiogenesis related factors, HIF-1α and VEGF. Received 28 February 2007; returned for revision 19 March 2007; accepted by J. Di Battista 11 July 2007     

乌梢蛇Ⅱ型胶原提取及其诱导大鼠关节炎特性的研究

提取和纯化乌梢蛇Ⅱ型胶原,观察乌梢蛇Ⅱ型胶原诱导大鼠关节炎的特性,为研究乌梢蛇治疗类风湿关节炎的机制提供研究方向。离心提取乌梢蛇Ⅱ型胶原,经DEAE-52纤维素交换柱纯化,斑点免疫渗滤试验和免疫印迹试验进行成分鉴定。乌梢蛇Ⅱ型胶原不完全佐剂免疫大鼠,观察大鼠关节炎发生情况、抗Ⅱ型胶原抗体、CD 4/CD8亚群、血清中TNF-α、IL-10、IL-1β、IL-4的变化。乌梢蛇Ⅱ型胶原提纯品在SDS-PAGE凝胶电泳上呈一条区带,斑点免疫渗滤试验和免疫印迹试验为阳性反应。诱导大鼠关节炎发生率为86.67%,关节炎大鼠血清中抗Ⅱ型胶原抗体明显增高(P<0.01),CD4 T细胞亚群和CD4~+/CD8~+增高(P<0.05),TNF-α含量增加(P<0.01),IL-10含量降低(P<0.01),血清中IL-1β和IL-4含量无变化。结果显示,乌梢蛇Ⅱ型胶原能通过免疫方法可以诱导大鼠产生多关节炎,并且体内有自身免疫反应的表现。     

LncRNA TMPO-AS1 up-regulates the expression of HIF-1α and promotes the malignant phenotypes of retinoblastoma cells via sponging miR-199a-5p

BackgroundLong non-coding RNA (lncRNA) TMPO antisense RNA 1 (TMPO-AS1) is reported to be oncogenic in prostate cancer and lung cancer. This study aims to investigate the expression and biological function of it in retinoblastoma (RB), and explore its regulatory role for miR-199a-5p and hypoxia-inducible factor-1α (HIF-1α).MethodsPaired RB samples were collected, and the expression levels of TMPO-AS1, miR-199a-5p and HIF-1α were examined by quantitative real-time polymerase chain reaction (qRT-PCR); TMPO-AS1 overexpressing plasmids and TMPO-AS1 shRNA were transfected into HXO-RB44 and SO-Rb50 cell lines respectively, and then proliferation, migration and invasion of RB cells were detected by CCK-8 assay and Transwell method. qRT-PCR and western blot were used to analyze the regulatory function of TMPO-AS1 on miR-199a-5p and HIF-1α; luciferase reporter gene assay was used to determine the regulatory relationship between miR-199a-5p and TMPO-AS1.ResultsTMPO-AS1 was significantly up-regulated in cancerous tissues of RB samples (relatively expression: 2.97 vs 3.93, p < 0.001), negatively correlated with miR-199a-5p (r=-0.4813, p < 0.01). There was one binding site on TMPO-AS1 for miR-199a-5p. After transfection of TMPO-AS1 shRNAs into RB cells, the proliferation, migration and invasion of cancer cells was significantly inhibited, while TMPO-AS1 had opposite effects; TMPO-AS1 was also demonstrated to regulate the expression of HIF-1α on both mRNA and protein levels via negatively regulating miR-199a-5p.ConclusionTMPO-AS1 is abnormally up-regulated in RB tissues, and it can modulate the proliferation and migration of RB cells. It has the potential to be the “ceRNA” to regulate HIF-1α expression by sponging miR-199a-5p.     

抗IL-1βscfv抗体与TNF-α可溶性受体融合蛋白的构建及其生物学活性分析

目的 设计原核表达抗人白细胞介素-1β单链抗体(IL-1βscfv)与TNF-α可溶性受体的融合蛋白,并分析其生物学活性,为类风湿关节炎的治疗提供新的候选药物.方法 利用RT-PCR方法从HeLa细胞总RNA中扩增了人肿瘤坏死因子Ⅰ型受体胞外区基因片段,与抗人IL-1βscfv通过抗体铰链区融合,克隆至pET27b(+)表达载体中,构建成pET-IL-1scfv:TNFR1质粒;转化大肠杆菌Rosetta进行表达.从包涵体中纯化得到IL-1scfv:TNFR1蛋白,进行IL-1scfv:TNFR1的鉴定及活性检测.结果 ELISA结果证明,IL-1scfv:TNFR1可以分别结合hIL-1β和hTNF-α,并呈现剂量依赖性,表明IL-1scfv:TNFR1的单链抗体部分和可溶性受体部分可以各自形成正确的空间构象;免疫斑点分析证明,IL-1scfv:TNFR1与hTNF-α结合后仍可以结合hIL-1β,具有同时结合hIL-1β和hTNF-α的两种靶分子的能力,表明IL-1scf:TNFR1的两个部分不会相互影响与靶分子的结合.活性测定结果表明,IL-1 scfv:TNFR1可以有效封闭肿瘤坏死因子对L929细胞的细胞毒效应,具有显著抑制hIL-1β促进L929细胞增殖的活性,IL-1scfv:TNFR1对两种靶分子的拮抗作用均呈现明显的剂量依赖性.结论 成功构建了能够同时结合hIL-1β和hTNF-α两种靶因子的双特异性IL-1 scfv:TNFR1融合蛋白,并能有效抑制hTNF-α和IL-1β的生物活性,为类风湿关节炎的治疗提供新的候选药物.     

HIF-1α和VEGF在宫颈癌中的表达及相关性研究

目的：探讨缺氧诱导因子-1α（HIF-1α）及血管内皮生长因子（VEGF）在宫颈癌发生发展过程中蛋白表达情况及两者的相关性。方法：应用免疫组织化学S-P方法检测10例正常宫颈（NCE）、18例宫颈上皮内瘤变（CIN）、77例宫颈癌（ICC）组织中HIF-1α和VEGF蛋白的表达情况。结果：在正常宫颈上皮、宫颈上皮内瘤变组织、宫颈癌（Ⅰ—ⅡA期）组织中，HIF．1d蛋白的阳性表达率分别是0．00％、33．33％、70．13％（P〈0．05），VEGF蛋白的阳性表达率分别为10．00％、44．44％、74．00％（P〈0．05）；且随HIF—1α表达增强，VEGF阳性表达率递增，两者呈正相关（P〈0．05）。结论：HIF-1α及VEGF的表达与宫颈癌的发生发展密切相关且两者呈正相关，HIF-1α蛋白可能以转录激活的形式上调VEGF基因的表达，诱导血管生成，促进了宫颈癌的发生发展。