胰岛素样生长因子Ⅰ和碱性成纤维细胞生长因子对人类牙乳头间充质细胞增殖和分化的影响**☆

背景：研究表明,胰岛素样生长因子Ⅰ和碱性成纤维细胞生长因子对细胞的增殖分化有重要作用,但对人类牙乳头间充质细胞生物学特性有何影响尚不清楚。 目的：观察胰岛素样生长因子Ⅰ和碱性成纤维细胞生长因子对体外培养人类牙乳头间充质细胞增殖和分化能力的影响。 方法：在建立人类牙乳头间充质细胞培养模型的基础上,①分别用含体积分数为1%或10%胎牛血清的DMEM/F12培养基将两种生长因子配制成不同的实验终浓度,取第4代生长良好的人类牙乳头间充质细胞,分别加入含0,0.1,1,10,100 μg/L碱性成纤维细胞生长因子和0,25,50,75,100 μg/L胰岛素样生长因子Ⅰ的培养基(0 μg/L作对照),培养96 h后四甲基偶氮唑盐法测增殖活性。②设10 μg/L碱性成纤维细胞生长因子组、100 μg/L胰岛素样生长因子Ⅰ组、碱性成纤维细胞生长因子+胰岛素样生长因子Ⅰ组和对照组。同上述方法每孔分别加含相应质量浓度生长因子的培养液,分别在培养1,3,5,7 d后,四甲基偶氮唑盐法测细胞增殖活性,改良酶动力学法测定细胞碱性磷酸酶活性。 结果与结论：在0~100 μg/L质量浓度范围时,两种生长因子对人类牙乳头间充质细胞增殖具有促进作用,碱性成纤维细胞生长因子的促增殖作用比胰岛素样生长因子Ⅰ大,联合作用时有协同促增殖作用。碱性成纤维细胞生长因子的最大有效质量浓度为10 μg/L,胰岛素样生长因子Ⅰ的最大作用质量浓度为100 μg/L。在0~7 d时,碱性成纤维细胞生长因子对人类牙乳头间充质细胞的碱性磷酸酶活性影响不明显,随时间的增加,胰岛素样生长因子Ⅰ对细胞碱性磷酸酶活性影响增加,与碱性成纤维细胞生长因子联用时,对增加碱性磷酸酶的活性有协同作用。     

胰岛素样生长因子1、血小板源性生长因子、转化生长因子β1对

背景：生长因子可调整椎间盘细胞的增殖、分化及基质代谢,用于椎间盘退变的生物学治疗,既往的研究偏重于成骨性生长因子对髓核细胞的作用,对纤维环细胞的研究相对较少。 目的：观察体外培养条件下胰岛素样生长因子1、血小板源性生长因子、转化生长因子β1对人退变纤维环细胞生物学活性的影响。 设计、时间及地点：对照观察实验,于2007-11/2008-12在解放军南京军区福州总医院完成。 材料：腰椎间盘手术患者的纤维环组织。 方法：结合酶消化法和组织块培养法,体外单层培养人退变纤维环原代细胞。传代后通过免疫组织化学染色鉴定细胞。对传3代细胞分别采用不同生长因子干预,分为 100 μg/L胰岛素样生长因子1组、10 μg/L 血小板源性生长因子组、10 μg/L 转化生长因子β1组、100 μg/L 胰岛素样生长因子 1+ 10 μg/L 血小板源性生长因子组、100 μg/L 胰岛素样生长因子1+ 10 μg/L 转化生长因子β1组、10 μg/L 血小板源性生长因子+10 μg/L 转化生长因子β1组、100 μg/L 胰岛素样生长因子1+10 μg/L血小板源性生长因子+10 μg/L 转化生长因子β1组,以不加生长因子为对照组。 主要观察指标：免疫组织化学染色观察传3代细胞。干预3,6 d后,采用MTT法测定传3代细胞增殖,ELISA法测定细胞培养上清液中Ⅰ、Ⅱ型胶原和聚集蛋白聚糖质量浓度。 结果：传3代细胞Ⅰ、Ⅱ型胶原免疫组织化学染色阳性。胰岛素样生长因子1促进人退变纤维环细胞增殖,轻度抑制细胞合成Ⅰ型胶原,促进合成Ⅱ型胶原和聚集蛋白聚糖,促Ⅱ型胶原合成作用轻度强于转化生长因子β1。血小板源性生长因子促进细胞增殖,作用强于胰岛素样生长因子1,其抑制细胞合成Ⅰ型胶原,轻度促进合成Ⅱ型胶原,无促合成聚集蛋白聚糖作用。转化生长因子β1抑制细胞增殖,促进合成Ⅰ、Ⅱ型胶原及聚集蛋白聚糖,促聚集蛋白聚糖合成作用强于胰岛素样生长因子1。多种因子联合作用较单种未见明显优势,未能呈现协同效应。 结论：胰岛素样生长因子1、血小板源性生长因子、转化生长因子β1明显改变人退变纤维环细胞的生物学活性,可应用于对人类椎间盘退变的生物学治疗。     

转化生长因子β1和胰岛素样生长因子Ⅰ及其协同作用对人皮肤成纤维细胞增殖的影响

背景：皮肤成纤维细胞注射移植为颜面部皱纹及微小凹陷等治疗提供了一种全新的方法,然而成纤维细胞的获取及体外快速扩增往往需要较长时间,以至于不能满足临床需要。 目的：观察转化生长因子β1、胰岛素样生长因子Ⅰ及两者协同作用对体外培养人皮肤成纤维细胞增殖的影响。 方法：人皮肤成纤维细胞原代培养,取第3代细胞种入96孔板,加入不同质量浓度的转化生长因子β1或胰岛素样生长因子Ⅰ作用于细胞,分别作用3,6,9 d采用MTT法检测细胞增殖情况。同法设4个组,分别为：阴性对照组、转化生长因子β1最佳效应浓度组、胰岛素样生长因子Ⅰ最佳效应浓度组、两者最佳效应浓度联用组,作用3,6,9 d采用MTT法检测增殖情况。 结果与结论：作用6 d和9 d,10.0 μg/L转化生长因子β1、50.0 μg/L胰岛素样生长因子Ⅰ组与对照组相比均有促细胞增殖效应(P < 0.05),此即为各生长因子最佳效应浓度。作用6 d和9 d,转化生长因子β1与胰岛素样生长因子Ⅰ各浓度实验组与相应对照组相比,差异均有显著性意义(P < 0.05)。10.0 μg/L转化生长因子β1与50.0 μg/L胰岛素样生长因子Ⅰ联合作用9 d可显著促进细胞增殖,与其他组相比,差异有显著性意义(P < 0.05)。提示转化生长因子β1、胰岛素样生长因子Ⅰ均可体外促进人皮肤成纤维细胞增殖,最佳效应浓度联用促增殖效果优于各自单独使用。     

转化生长因子β1对长波紫外线照射皮肤成纤维细胞胰岛素样生长因子1、角质形成细胞生长因子、血管内皮细胞生长因子表达的影响*☆

背景：研究认为衰老和光老化状态下生长因子的表达不一定一致,生长因子与皮肤老化的关系越来越受到重视。 目的：验证转化生长因子β1对长波紫外线照射体外培养的皮肤成纤维细胞胰岛素样生长因子1、角质细胞生长因子、血管内皮细胞生长因子表达的影响 。 设计、时间及地点：以体外培养细胞为观察对象,随机重复实验,于2006-07/2008-03在解放军第四军医大学全军整形外科研究所进行。 材料：成纤维细胞来自解放军第四军医大学整形外科门诊患者包皮,采用组织块法培养获得。 方法：通过酶联免疫法(ELISA)使用不同剂量长波紫外线 (0,5 ,10,20 J/ cm2)照射体外培养的皮肤成纤维细胞,测定上清液中胰岛素样生长因子1、角质形成细胞生长因子、 血管内皮细胞生长因子的质量浓度;以长波紫外线15 J/ cm2为照射剂量,选择不同剂量转化生长因子β1即小剂量组(0.1 μg/L)、中剂量组(1 μg/L)、大剂量组(10 μg/L)对培养的皮肤成纤维细胞进行干预。 主要观察指标：不同剂量长波紫外线照射、以及转化生长因子β1处理后皮肤成纤维细胞胰岛素样生长因子1、血管内皮细胞生长因子、角质形成细胞生长因子的表达。 结果：长波紫外线照射体外培养的皮肤成纤维细胞导致胰岛素样生长因子1、角质形成细胞生长因子分泌水平下降,血管内皮细胞生长因子分泌升高;转化生长因子β1处理后,转化生长因子β1剂量依赖性地提高长波紫外线照射体外培养的皮肤成纤维细胞分泌的3种细胞因子的水平。转化生长因子β1大剂量组胰岛素样生长因子1、角质形成细胞生长因子、血管内皮细胞生长因子含量明显高于长波紫外线照射组(P < 0.01)。 结论：长波紫外线照射抑制体外培养成纤维细胞中胰岛素样生长因子1、角质形成细胞生长因子表达, 促进血管内皮细胞生长因子表达。转化生长因子β1可提高长波紫外线照射体外培养的皮肤成纤维细胞分泌胰岛素样生长因子1、角质形成细胞生长因子、血管内皮细胞生长因子3种细胞因子的水平,有利于真皮内结构的恢复和重建。     

体外培养软骨细胞与生长因子的加入*

背景：自体软骨细胞体外培养生长缓慢，极易发生去分化，胰岛素样生长因子和碱性成纤维细胞生成因子可以刺激关节软骨细胞的增殖和分化。 目的：观察应用碱性成纤维细胞生成因子和胰岛素样生长因子1对关节软骨细胞的作用。 方法：采用成人关节软骨细胞体外培养，以碱性成纤维细胞生成因子(10 μg/L)和胰岛素样生长因子1(100 mg/L)对其作用，采用MTT法和免疫细胞化学技术进行观察和评价。 结果与结论：在体外培养软骨细胞时应用碱性成纤维细胞生成因子可以明显促进细胞增殖，但同时发生去分化。在体外培养软骨细胞时应用胰岛素样生长因子1可以明显促进细胞产生细胞外基质，有利于软骨细胞表型的表达，对细胞的增殖作用不明显。提示采用顺序性应用胰岛素样生长因子1和碱性成纤维细胞生成因子对体外培养的软骨细胞进行作用，可以在更短的时间内获得大量的、具有良好细胞表型的软骨细胞。 关键词：碱性成纤维细胞生长因子；胰岛素样生长因子1；软骨细胞；培养；体外     

生长因子对骨骼肌肌源性干细胞增殖的影响★

背景：单纯应用差速贴壁法获得的原代肌源性干细胞数量少，生长速度较慢，不利于获取满足实验及将来临床所需的细胞数量。 目的：观察碱性成纤维细胞生长因子和胰岛素样生长因子1在体外对体外培养肌源性干细胞增殖的作用，探索体外大量培养肌源性干细胞的方法和条件。 设计、时间及地点：观察对比实验，于2007-09/2008-04在武汉大学人民医院泌尿外科实验室完成。 材料：2周龄体质量50~80 g的SD大鼠20只。 方法：采用改良差速贴壁培养法分离大鼠骨骼肌肌源性干细胞：取新生大鼠肌肉标本分离细胞；维持总的贴壁时间基本不变，减少贴壁次数，取第2代肌源性干细胞进行体外成肌分化。 主要观察指标：以免疫细胞化学法及免疫荧光法鉴定肌源性干细胞。以MTT 比色实验检测5，10，20，40，80 μg/L的碱性成纤维细胞生长因子、胰岛素样生长因子1及胰岛素样生长因子1+碱性成纤维细胞生长因子对肌源性干细胞增殖的影响。以流式细胞仪检测碱性成纤维细胞生长因子、胰岛素样生长因子1 和胰岛素样生长因子1+碱性成纤维细胞生长因子对细胞周期的影响。 结果：肌源性干细胞原代培养70 h后开始贴壁，呈规则的圆形。1周后细胞数量增多，细胞展开呈纺锤性。免疫细胞化学染色法和细胞免疫荧光法显示肌源性干细胞呈结蛋白、干细胞抗原1和CD34阳性。传代细胞加入碱性成纤维细胞生长因子、胰岛素样生长因子1后肌源性干细胞的增殖活性增强，两种细胞因子作用强度相近，随着两种细胞因子浓度的提高促细胞增殖作用逐渐提高,在达到有效浓度后作用趋于饱和。两种细胞因子作用机制不同，碱性成纤维细胞生长因子主要是增加G2M期的细胞比例，通过缩短细胞周期达到增殖作用, 胰岛素样生长因子1则主要增加S期细胞的比例，通过活化由G0期到G1期转化的进入有丝分裂细胞来达到增殖作用。 结论：胰岛素样生长因子1 和碱性成纤维细胞生长因子都具有刺激肌源性干细胞增殖的作用，两者联合作用既可以增加早期由G0期进入G1期的数量，又可以减短细胞有丝分裂周期，从而达使促增殖效果更快、更强。     

胰岛素样生长因子Ⅰ对豚鼠巩膜成纤维细胞增殖的影响☆

目的：研究表明，体内一定浓度的生长因子是保证眼球正常发育的必要条件，过少或过量都会引起巩膜的异常改变。实验拟进一步验证胰岛素样生长因子Ⅰ对体外培养的豚鼠巩膜成纤维细胞生长的影响。 方法：实验于2006-09/2007-03在郑州大学基础医学院省肿瘤病理学重点实验室完成。①实验材料：出生3 d内3色豚鼠后部巩膜组织由郑州大学基础医学院动物实验中心提供；实验用胰岛素样生长因子Ⅰ 为Sigma公司产品。②实验过程及评估：采用器官组织块培养法获取原代巩膜成纤维细胞，选取传3，4代的豚鼠巩膜成纤维细胞用于实验；采用倒置相差显微镜下观察细胞形态并采用免疫化学染色法进行细胞鉴定；应用四甲基偶氮唑盐比色法和流式细胞仪分析技术,观察不同质量浓度胰岛素样生长因子Ⅰ (0.1，0.5，5，10，50 μg/L )对体外培养的豚鼠巩膜成纤维细胞增殖和细胞生长周期的影响。 结果：①巩膜成纤维细胞鉴定：倒置相差显微镜下观察,细胞透亮呈梭形，在细胞密度较低时细胞排列成网状,密度较高时呈束状；波形蛋白染色阳性。胞浆内可见棕黄色阳性反应产物, 证实所培养的细胞为巩膜成纤维细胞。②体外培养的豚鼠巩膜成纤维细胞增殖和细胞生长周期：0.5，5，10 μg/L的胰岛素样生长因子Ⅰ能促进豚鼠巩膜成纤维细胞增殖,呈剂量依赖性(F = 84.510，P =0.000 < 0.05)，最佳质量浓度为10 μg/L。流式细胞仪结果显示, 胰岛素样生长因子Ⅰ处理48 h后,与对照组比较，巩膜成纤维细胞G0~G1期百分比降低（78.2%下到64.5%，P < 0.05）,而S期百分比显著升高（10.4%上升到16.4%，P < 0.05）,增殖指数百分比增高（27.8%增高到56.5 %，P < 0.05）。 结论： 在一定浓度范围内外源性胰岛素样生长因子Ⅰ可以促进体外培养的豚鼠巩膜成纤维细胞增殖。     

胰岛素样生长因子Ⅰ对组织工程肌腱细胞增殖影响的量效关系★

目的: 胰岛素样生长因子Ⅰ是一种潜在的促有丝分裂剂，对肌腱细胞有促进分裂增殖的作用。实验将不同剂量胰岛素样生长因子Ⅰ作用于第2代肌腱细胞，进一步验证其对细胞增殖的影响并探讨其量效关系。 方法：实验于2004-11/2005-04在天津医院实验室完成。①实验材料：天津医院实验室提供的精选法国罗曼鸡受精鸡蛋200只，在孵育19 d时，随机取出10只鸡胚肌腱。②实验过程及分组：分离培养肌腱细胞，观察肌腱细胞形态及生长规律。取第2代肌腱细胞接种于六孔板，分别给予不同浓度的胎牛血清，观察血清浓度对肌腱贴壁及生长的影响。取第2代肌腱细胞接种于96孔板，分为7组。前5组分别加入含不同剂量胰岛素样生长因子Ⅰ（1，5，10，50，200 μg/L）的体积分数为0.02的胎牛血清培养液；第6组加入体积分数为0.05的胎牛血清培养液做为阳性对照，第７组加入体积分数为0.02的胎牛血清培养液做为阴性对照。③实验评估：采用四甲基偶氮唑盐法及瑞士－姬姆萨染色观察不同剂量的胰岛素样生长因子Ⅰ对细胞增殖的影响。 结果：①肌腱细胞贴壁后生长很快，原代细胞1周左右即可传代，增殖速度与营养液中胎牛血清浓度呈正相关。②肌腱细胞增殖速度随胰岛素样生长因子Ⅰ剂量加大而有增加趋势。第２天、第4天，胰岛素样生长因子Ⅰ1 μg/L组、5 μg/L组和阴性对照组之间差异无显著性(P > 0.05)；胰岛素样生长因子Ⅰ10 μg/L组、50 μg/L组和200 μg/L组之间差异无显著性(P > 0.05)。②第2天，阳性对照组增殖高于胰岛素样生长因子Ⅰ1 μg/L组与5 μg/L组，低于10 μg/L组、50 μg/L组和200 μg/L组，差异有显著性(P < 0.000或< 0.006)；第4天，阳性对照组增殖高于胰岛素样生长因子Ⅰ各剂量组和阴性对照组，差异有显著性(P < 0.000或< 0.006)。③第2天、第4天，胰岛素样生长因子Ⅰ1μg/L组、5μg/L组和阴性对照组低于10μg/L组，50μg/L组，200μg/L组，差异有显著性(P < 0.000)；阳性对照组增殖高于阴性对照组，统计分析差异有显著性(P < 0.000)。 结论：胰岛素样生长因子Ⅰ对肌腱细胞增殖的促进作用，随胰岛素样生长因子Ⅰ浓度增高而有增高趋势，10 μg/L为其较适合浓度，同时发现培养血清浓度对肌腱细胞有其特殊作用，浓度越高，肌腱细胞贴壁越快，并对增殖有明显促进作用。     

蛇床子素干预人增生性瘢痕成纤维细胞增殖及转化生长因子β1的表达

背景：蛇床子素对体外培养人增生性瘢痕成纤维细胞增殖和细胞分泌的转化生长因子β1有抑制作用,但其具体作用机制尚待进一步研究。 目的：体外观察蛇床子素对人增生性瘢痕成纤维细胞增殖以及对细胞转化生长因子β1的影响。 方法：体外原代培养人增生性瘢痕成纤维细胞,以不同浓度的蛇床子素作用于成纤维细胞,观察细胞形态的变化,应用MTT法和生长曲线法检测蛇床子素对细胞增殖活性的影响。免疫组织化学检测细胞转化生长因子β1的表达。 结果与结论：蛇床子素能明显抑制人增生性瘢痕成纤维细胞的生长。MTT法检测的IC50为(15.2±2.0) μmol/L,可以明显下调细胞转化生长因子β1的表达(P < 0.05)。说明蛇床子素对人增生性瘢痕成纤维细胞有很强的生长抑制作用并可以下调转化生长因子β1的表达。     

碱性成纤维细胞生长因子对生长板软骨细胞增殖与分化的影响

背景: 体外培养软骨细胞经历多次传代后，其增殖能力将逐渐退化。实验表明，碱性成纤维细胞生长因子可促进软骨细胞增殖，并可能影响软骨细胞的表型与分化。 目的：观察碱性成纤维细胞生长因子对生长板软骨细胞增殖和分化的影响，并筛选其用于体外软骨细胞培养的最佳剂量。 设计、时间及地点：以细胞为观察对象的观察对比实验，于2004-10/2005-02在暨南大学医学院生化教研室组织工程实验室完成。 材料：选用12只新西兰白兔用于分离软骨细胞，碱性成纤维细胞生长因子由英国PeproTech公司生产。 方法：分离并在低血清条件下培养兔生长板软骨细胞。根据实验需要加入0，1，2.5，5，10，25，50及100 μg/L 8个剂量的碱性成纤维细胞生长因子，进行各项指标观察。 主要观察指标：应用改良四甲基偶氮唑蓝比色法检测细胞增殖倍数；应用羟脯氨酸法测定软骨细胞胶原产量；应用酶动力学方法测定碱性磷酸酶活性。 结果：①碱性成纤维细胞生长因子剂量在5~100μg/L范围内可以促进软骨细胞增殖，并以25 μg/L时刺激效果最为显著。②当碱性成纤维细胞生长因子剂量高于 25 μg/L时，软骨细胞的胶原合成被抑制。③当碱性成纤维细胞生长因子剂量高于 1 μg/L时，软骨细胞的碱性磷酸酶活性被抑制。 结论：碱性成纤维细胞生长因子可以刺激生长板软骨细胞增殖，并在剂量高于25 μg/L时抑制生长板软骨细胞的分化。     

RNA interference affects tumorigenicity and expression of insulin-like growth factor-1, insulin-like growth factor-1 receptor, and basic fibroblast growth factor-2 in rat C6 glioma cells

BACKGROUND： Human gliomas are more likely to express basic fibroblast growth factor-2 （FGF-2） insulin-like growth factor-1（IGF-1）, and IGF-1 receptor （IGF-1R） than normal brain tissue. These factors activate signal transduction systems of Ras/MAPK and PI3K/Akl, which promote glioma growth. OBJECTIVE： To utilize RNA interference （RNAi） technique to down-regulate FGF-2, IGF-1, and IGF-1R gene expression, and to investigate the effects of these genes on rat C6 glioma cells, as well as the feasibility of RNAi for treating glioma. DESIGN, TIME AND SETTING： This neurooncological, randomized, controlled, in vivo and in vitro experiment, which used RNAi methodology, was performed at the Laboratory of Molecular Biology, Institute of Biochemistry, Chinese Academy of Sciences between August 2005 and February 2008. MATERIALS： Rat C6 cell lines were purchased from Shanghai Institute of Cellular Biology Affiliated to Chinese Academy of Sciences. Small interfering RNA （siRNA） was synthesized by Shanghai GenePharma. Anti-IGF-1, anti-IGF-1R, anti-FGF-2, anti-mouse and anti-rabbit IgG G1-HRP antibodies were provided by Santa Cruz Biotechnology, USA. Four to six week-old BALB/c nude mice were purchased from the Laboratory Animal Center, Chinese Academy of Sciences. METHODS： C6 glioma cells were transfected with siRNA, which was chemically synthesized in vitro to correspond to endogenous FGF-2, IGF-1, and IGF-1R genes. The inhibition ratio of targeting mRNA expression was detected by semiquantitative RT-PCR, and protein expression was determined by Western blot analysis. C6 glioma cell proliferation was observed using a growth curve C6 glioma cell apoptosis rate and cell cycle were detected by flow cytometry. C6 glioma cell growth regression was observed by transwell migration assay. In addition, nude mouse subcutaneous tumor models were used in this study. For studying the anti-tumor effects of IGF-1 and IGF-1R siRNA, two blank control groups, with six mice each, were set up： A （2.5 μg siRNA was injected one week after C6 cells were inoculated, Le., when tumor volume reached 8 mm × 8 mm） and B （siRNA was injected at the same time with C6 cells were inoculated. To study the effects of FGF-2 siRNA, the groups consisted of a blank control group, negative control group, 2.6 μg siRNA group, 4 μg siRNA group, and 5.3 μg siRNA group, with six mice each. MAIN OUTCOME MEASURES： mRNA and protein inhibition ratio of FGF-2, IGF-1, and IGF-1 R; C6 glioma cell proliferation, apoptosis, and cycle growth arrest; C6 glioma cell growth regression and subcutaneous tumorigenicity rates. RESULTS： All siRNA constructs proved to be effective. After 48 hours, transfection of 200 nmol/L siRNA resulted in a FGF-2 or IGF-1R gene inhibition ratio 〉 80% and an IGF-1 gene inhibition ratio of approximately 70%. Protein expression levels for FGF-2, IGF-1, and IGF-1R decreased in a dose-dependent manner following siRNA transfection, with an inhibition rate 〉 85%, 60%, and 50%, respectively. C6 glioma cell proliferation and apoptosis rates increased in proportion to siRNA. The apoptosis rate of C6 glioma cells induced by FGF-2, IGF-1, and IGF-1R siRNA was 39.96%, 15.07% and 22.47%, respectively （P 〈 0.01）. Transfection of 200 nmol/L IGF or IGF-1R siRNA for 48 hours suppressed C6 glioma cell migration. At 30 days after intratumoral injection of 2.6, 4, and 5.3 tJg FGF-2 siRNA, tumor growth regression rate of FGF-2 siRNA was 56%, 67%, and 86%, respectively. The tumor growth regression rate was 71.88% and 45.71%, respectively, when IGF-1 or IGF-1R siRNA was intratumorally injected 1 week after C6 glioma cell transplantation. When IGF-1 or IGF-1 R siRNA was intratumorally injected during C6 glioma cell transplantation, the tumor growth regression rate was 78.13% and 74.29%, respectively. CONCLUSION： siRNA transfection downregulated gene expression of FGF-2, IGF-1, and IGF-1R In addition, siRNA treatment markedly suppressed glioma cell proliferation, growth, and migration, and concomitantly reduced subcutaneous tumorigenicity.     

急性脑梗死患者血清TGF—β1检测的临床意义

目的探讨血清转化生长因子－β１（ＴＧＦ－β１）在急性脑梗死（ＡＣＩ）发病过程中的变化及其意义。方法采用酶联免疫吸附试验动态检测３５例ＡＣＩ患者血清ＴＧＦ－β１浓度，分析其与梗死部位、大小、病情的相关性。结果ＡＣＩ患者血清ＴＧＦ－β１浓度急性期降低，恢复期升高。皮质梗死组ＴＧＦ－β１高于皮质下组（Ｐ〈０．０５）；梗死体积大则ＴＧＦ－β１浓度市（Ｐ〈０．０５）；重型组ＴＧＦ－β１浓度高于轻型组，但无统     

Fibroblast growth factor-2 deficiency causes defects in adult hippocampal neurogenesis, which are not rescued by exogenous fibroblast growth factor-2

Neurogenesis within the adult brain is restricted to selected areas, one of which is the dentate gyrus (DG). Several growth factors have been reported to affect neurogenesis in the adult DG. However, a role of fibroblast growth factor-2 (FGF-2) in adult hippocampal neurogenesis has not been firmly established. We have analyzed neurogenesis in the DG using in vivo and in vitro approaches. FGF-2(-/-) mice revealed no alterations in the number of proliferating cells but a significant decrease in the numbers of newly generated neurons. Moreover, FGF-2 added to hippocampal slice cultures from FGF-2(-/-) mice was unable to rescue the phenotype. Although an increase in death of neurogenic cells in the FGF-2-deficient DG could not be specifically demonstrated, there was a massive increase in global cell death in FGF-2(-/-) hippocampal slice cultures compared with slices from wild-type mice. Cell death could not be prevented by addition of FGF-2. Neutralization of endogenous FGF-2 in hippocampal slices did not interfere with neurogenesis in a short-term paradigm. Together, our data suggest that FGF-2 is essentially required for maturation of new neurons in adult hippocampal neurogenesis but is likely to operate synergistically in combination with other mechanisms/growth factors.     

Fibroblast growth factor-2 enhances functional recovery of reinnervated muscle

Long-term denervation of muscles results in fibrosis and fat replacement, which prevent muscles from regaining contractile function despite reinnervation. Therefore, prevention of muscle atrophy between nerve repair and muscle reinnervation may improve the functional outcome. A variety of growth factors play significant roles in muscle mass modulation and muscle regeneration. The purpose of the present study was to investigate the effect of fibroblast growth factor-2 (FGF2) and nerve growth factor (NGF) on muscle mass modulation after denervation and reinnervation using a nerve-to-muscle neurotization model. Growth factors were injected into the anterior tibial muscle after direct neurotization of muscles every 7 days up to 4 weeks after surgery. FGF2 significantly increased the amplitude of compound muscle action potentials (CMAPs), wet muscle weight, and the number of motor endplates, especially at higher concentration, compared to the vehicle. In contrast, NGF did not increase either the amplitude of CMAPs or muscle weight, although it significantly increased the number of motor endplates. These findings indicate that both growth factors enhance reinnervation of muscles; however, only FGF2 is involved in muscle mass modulation.     

Association of insulin-like growth factor-1 receptor polymorphism in dementia

There is an increasing interest in how oxidative stress can cause cells to go into apoptosis in both normal ageing and in neurodegenerative disorders. Previous research has implicated insulin-like growth factor-1 (IGF-1) as being involved in the pathogenesis in Alzheimer's disease (AD) by protecting the neurons through reducing neuronal susceptibility to oxidative stress. IGF-1 receptor (IGF-1R) polymorphisms alter cerebral and systemic levels of IGF-1 and may alter the function of the receptor. We genotyped the IGF-1R gene by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) to assess whether this gene polymorphism can be linked to dementia. We used leukocyte DNA from 72 patients with AD, 75 patients with vascular dementia (VaD), 14 patients with mixed dementia (AD+VaD), and a control group consisting of 209 individuals without a history of progressive neurological disorders. Analysis of gene frequency for gender revealed a significant difference between female VaD patients and female controls carrying at least one A allele (OR = 1.8, CI 95% 1.1-2.9, p = 0.02), but not for male patients. In addition, we found a strong tendency to a difference between all cases of female dementia patients and controls carrying the A allele (OR = 1.5, CI 95% 0.99-2.2, p = 0.054). Our results suggest that the A allele of IGF-1R may be involved in the pathogenesis of VaD in females.     

Angiogenic and inflammatory responses following skeletal muscle injury are altered by immune neutralization of endogenous basic fibroblast growth factor, insulin-like growth factor-1 and transforming growth factor-β1

Injured skeletal muscle degeneration comprises early microvascular changes and inflammatory cell infiltration, possibly under the control of several growth factors. We have studied the role of basic fibroblast growth factor (bFGF), insulin-like growth factor-1 (IGF1), and transforming growth factor beta-1 (TGFβ1), by injecting specific anti-growth factor neutralizing antibodies into mouse extensor digitorum longus muscle at the time of injury (denervation and devascularization). Four days later, at the height of damaged myofiber phagocytosis, we assessed quantitatively revascularization, phagocytic activity, and inflammation. The immune neutralization of bFGF reduced the number of capillaries, macrophages and mast cells, and delayed necrotic myofiber phagocytosis. The immune neutralization of IGF1 or TGFβ1 promoted muscle revascularization, macrophage infiltration and necrotic myofiber phagocytosis. While IGF1 neutralization reduced the number of mast cells and did not modify that of T-cells or neutrophils, TGFβ1 neutralization increased the number of all of these cells. This study strongly suggests differing roles for bFGF, IGF1 and TGFβ1 in angiogenic and inflammatory responses during muscle degeneration, apart from their known effects on the behaviour of myogenic cells.     

Increased circulating insulin-like growth factor-1 in late-onset Alzheimer's disease

BACKGROUND: Insulin-like growth factor (IGF)-1 has been implicated in the pathogenesis of Alzheimer's disease (AD). METHODS: We compared the level of circulating total and bioavailable IGF-1, by simultaneous measurements of IGF-1 and IGF binding protein (IGFBP)-3, between 87 patients diagnosed with AD and 126 age and sex matched control subjects without cognitive impairment. Blood samples were collected and IGF-1 and IGFBP-3 measured by ELISA. Subjects were also genotyped for apolipoprotein E. RESULTS: Total circulating IGF-1 levels were significantly raised in the AD group as compared to the control group (p=0.022). There was no significant difference in the circulating level of IGFBP-3 between the two groups. When the IGF-1 levels were ratioed against IGFBP-3 levels as an indicator of unbound, bioavailable circulating IGF-1, there was a significant increase in the molar IGF-1:IGFBP-3 ratio in the AD subjects (0.181 +/- 0.006) as compared to the controls (0.156 +/- 0.004) (p< 0.001). Logistic regression analysis revealed that an increase in the IGF-1:IGFBP-3 molar ratio increased the risk of AD significantly. CONCLUSION: The results of increased total and free circulating IGF-1 support the hypothesis that in its early stages late-onset AD reflects a state of resistance to IGF-1.     

Insulin and insulin-like growth factor-1 abnormalities in antipsychotic-naive schizophrenia

OBJECTIVE: The purpose of this study was to examine the evidence for the insulin-like growth factor-1 (IGF-1) deficiency hypothesis in the pathogenesis of schizophrenia. METHOD: The authors examined the fasting plasma levels of glucose, insulin, IGF-1, and cortisol in antipsychotic-naive schizophrenia patients (N=44) relative to age- and sex-matched healthy comparison subjects (N=44). Patients and comparison subjects were also matched for anthropometric measures and physical activity. RESULTS: Schizophrenia patients had a significantly higher mean plasma insulin level as well as a significantly higher mean insulin resistance score relative to healthy comparison subjects. The mean plasma IGF-1 level was significantly lower in patients. IGF-1 levels had a significant negative correlation with plasma insulin levels. The total positive symptoms score as well as the hallucinations subscore had a significant inverse relationship with IGF-1 levels. CONCLUSIONS: Deficient IGF-1 might underlie insulin resistance in schizophrenia. The IGF-1 deficit in antipsychotic-naive schizophrenia patients and its significant correlation with psychopathology scores suggest that IGF-1 might be potentially involved in the pathogenesis of schizophrenia.