慢性间断性缺氧对雄仔鼠肝胰岛素样生长因子-1基因组蛋白修饰的作用

目的:研究慢性间断性缺氧(CIH)时新生雄仔鼠肝形态学改变及胰岛素样生长因子-l(IGF-1)基因表达,探讨其表观遗传学分子机制.方法:将怀孕SD大鼠分为正常呼吸组(对照组)和间断缺氧组(缺氧组),建立CIH大鼠模型.分娩后取对照组和缺氧组1 d雄仔鼠肝组织,用电子显微镜观察肝组织的超微结构,免疫蛋白印迹和免疫组织化学...     

胰岛素样生长因子1和胰岛素对人a1（I）前胶原启动子活性的影响

目的：探讨胰岛素样生长因子1（IGF-1）和胰岛素对人a1(I)前胶原（COL1A1）基因启动子的调控作用及肿瘤坏死因子a(TNFa）、干扰素γ（IFNγ）、干扰素a(INFa）对这一作用的影响。方法：构建含 COL1A1基因启动子序列与氯霉素乙酰基转移酶（CAT）报告基因的重组体pCOLH0.27与pCOLH2.5,它们分别含该启动子序列-268～ 42bp(0.27kb)与-2483～ 42bp(2.5kb)片段。将这2个重组体瞬时转染人皮肤成纤维细胞,加入IGF-1或胰岛素,部分细胞还加入TNFa或IFNγ或IFNa,测定CAT活性。结果：13nmol/L IGF-1使转染后的这2个重组体活性分别增高3.68倍与4.04倍。2.5umol/L的胰岛素亦使之分别增高3.69倍与3.93倍。TNFa、IFNγ、IFNa能降低IGF-1与胰岛素诱导的pCOLH2.5活性。结论：IGF-1和胰岛素能在转录水平促进COL1A1基因的表达,TNF-a、INFγ、IFNa能抑制IGF-1和胰岛素的这一作用。     

胰岛素样生长因子1和胰岛素对人α1(Ⅰ)前胶原启动子活性的影响1

目的探讨胰岛素样生长因子1(IGF-1)和胰岛素对人α1(Ⅰ)前胶原(COL1A1)基因启动子的调控作用及肿瘤坏死因子α(TNFα)、干扰素γ(IFNγ)、干扰素α(IFNα)对这一作用的影响.方法构建含COL1A1基因启动子序列与氯霉素乙酰基转移酶(CAT)报告基因的重组体pCOLH0.27与pCOLH2.5,它们分别含该启动子序列-268～+42bp(0.27kb)与-2483～+42bp(2.5kb)片段.将这2个重组体瞬时转染人皮肤成纤维细胞,加入IGF-1或胰岛素,部分细胞还加入TNFα或IFNγ或IFNα,测定CAT活性.结果13nmol/LIGF-1使转染后的这2个重组体活性分别增高3.68倍与4.04倍.2.5μmol/L的胰岛素亦使之分别增高3.69倍与3.93倍.TNFα、IFNγ、IFNα能降低IGF-1与胰岛素诱导的pCOLH2.5活性.结论IGF-1和胰岛素能在转录水平促进COL1A1基因的表达,TNF-α、IFNγ、IFNα能抑制IGF-1和胰岛素的这一作用.     

胰岛素样生长因子1和胰岛素对人α1(Ⅰ)前胶原启动子活性的影响

目的 :探讨胰岛素样生长因子 1(IGF 1)和胰岛素对人α1(Ⅰ )前胶原 (COL1A1)基因启动子的调控作用及肿瘤坏死因子α(TNFα)、干扰素γ(IFNγ)、干扰素α(IFNα)对这一作用的影响。方法 :构建含COL1A1基因启动子序列与氯霉素乙酰基转移酶 (CAT)报告基因的重组体pCOLH0 2 7与pCOLH2 5 ,它们分别含该启动子序列 2 68～ 4 2bp( 0 2 7kb)与 2 4 83～ 4 2bp( 2 5kb)片段。将这 2个重组体瞬时转染人皮肤成纤维细胞 ,加入IGF 1或胰岛素 ,部分细胞还加入TNFα或IFNγ或IFNα,测定CAT活性。结果 :13nmol/LIGF 1使转染后的这 2个重组体活性分别增高 3 68倍与 4 0 4倍。 2 5 μmol/L的胰岛素亦使之分别增高 3 69倍与 3 93倍。TNFα、IFNγ、IFNα能降低IGF 1与胰岛素诱导的pCOLH2 5活性。结论 :IGF 1和胰岛素能在转录水平促进COL1A1基因的表达 ,TNF α、IFNγ、IFNα能抑制IGF 1和胰岛素的这一作用。     

慢性间歇性缺氧对认知功能及海马CA1区IGF-1表达的影响

目的:探讨慢性间歇性缺氧对SD大鼠认知功能及海马CA1区IGF-1表达的影响。方法:选取16只SD成年雄性大鼠,随机分为空白组(unhandled control group,UC)和慢性间歇性缺氧组(chronic intermittent hypoxia group,CIH),UC组正常饲养,CIH组每日间歇缺氧7小时,建立慢性间歇性缺氧模型,8周结束实验,所有大鼠在实验结束后进行Morris水迷宫测试,检测其学习和记忆能力,并利用免疫组化及图像分析测定大鼠海马CA1区IGF-1蛋白的表达。结果:①Morris水迷宫学习成绩(定位航行实验):第5天训练结束时,CIH组大鼠逃避潜伏期明显长于UC组,两组之间差异有统计学意义(P<0.05);②Morris水迷宫记忆成绩:CIH组的穿越平台次数(1.38±0.92)次较UC组显著减少[(3.75±1.04),P<0.01];CIH组在跨越目标象限时间占整个游泳的时间百分率(20.52±3.41)也较UC组显著减少[(39.89±5.63),P<0.01];③CIH组大鼠海马CA1区IGF-1蛋白的表达较UC组减少(P<0.05);④大鼠海马CA1区IGF-1表达和认知功能损害程度呈负相关(r=0.867,P<0.01)。结论:①慢性间歇性缺氧可导致大鼠认知功能下降;②慢性间歇性缺氧可导致大鼠海马CA1区IGF-1的表达减少;③IGF-1对慢性间歇性缺氧大鼠认知功能损害可能具有保护作用。     

人胰岛素样生长因子Ⅱ基因P1、P3启动子克隆及意义

目的:人胰岛素样生长因子Ⅱ(IGF-Ⅱ)基因P1、P3启动子的克隆。方法:根据GenBank数据库提供的IGF-Ⅱ基因DNA全序列及有关文献, 应用巢式PCR技术从L-02正常人胎肝细胞系中扩增分离出P1、P3启动子片段, 并采用TOPO TA Cloning kit将PCR产物克隆入pCR2.1-TOPO T载体。结果:经琼脂糖凝胶电泳及直接测序鉴定, 克隆的IGF-Ⅱ基因P1、P3启动子片段碱基序列与GenBank数据库一致。结论:成功克隆了IGF-II基因P1、P3启动子。     

KIR3 DL1启动子区域 CpG 岛甲基化对抗原表达的影响

目的：探索KIR3DL1启动子区域CpG岛甲基化对抗原表达的影响。方法选择抗原高表达KIR3DL1倡01502和低表达KIR3DL1倡005样本,分别测定启动子区域碱基序列和启动子区CpG岛甲基化程度。对携带有KIR3DL1倡01502、KIR3DL1倡005的NK细胞分别采用5-aza进行去甲基化处理,利用流式细胞仪检测KIR3DL1抗原。结果 KIR3DL1倡01502和KIR3DL1倡005启动子区域－65和－269位存在碱基差异,这导致它们的启动子区域有两个不同的CpG岛。 KIR3DL1倡01502启动子区CpG岛高度甲基化,去甲基化后相应的细胞表面抗原表达显著增加。而携带KIR3DL1倡005的细胞去甲基化处理后,抗原表达没有明显变化。结论 KIR3DL1倡01502启动子区域CpG岛甲基化影响抗原的表达,但是不同抗原表达模式的KIR3DL1等位基因可能存在不同的调控机制。     

胰岛素样生长因子1的体内表达对BXSB小鼠发病影响的研究

目的 初步研究人胰岛素样生长因子1(hIGF-1)对系统性红斑狼疮ＢＸＳＢ小鼠发病的影响。方法 用肌肉介导的电脉冲转基因技术,使得含有hIGF-1cDNA的重组真核表达质粒在小鼠体内表达,然后系统检测自身免疫相关性实验指标。结果 虽然hIGF-1促进雄性ＢＸＳＢ小鼠ＩgG类型抗ＤＮＡ抗体的产生,但是明显阻止其尿蛋白浓度的继续升高,抑制巨噬细胞分泌IL-1以及降低脾脏重量。结论 IGF-1具有减缓雄性BXSB小鼠狼疮性肾炎的作用。其作用机制之一可能与巨噬细胞分泌炎性介质的功能下降有关。     

脐血中胰岛素、胰岛素样生长因子-I及胰岛素样生长因子结合蛋白-2与胎儿生长受限的关系

目的探讨脐血中胰岛素、胰岛素样生长因子-I及胰岛素样生长因子结合蛋白-2的水平与胎儿生长受限的关系及意义。方法收集2011年7月～2013年5月在本院分娩的单胎足月正常妊娠（无产科并发症）孕妇及其新生儿各120例,根据出生体重将新生儿分为小于胎龄儿（SGA）组、适于胎龄儿（AGA）2组。胎儿娩出后即抽脐静脉血5ml,标本收集后用高效液相色谱法测定脐血中胰岛素、胰岛素样生长因子-I及胰岛素样生长因子结合蛋白-2的水平。结果SGA组脐带血清胰岛素、胰岛素样生长因子-I水平明显低于AGA组,差异均有统计学意义（P〈0.01）;SGA组脐血胰岛素样生长因子结合蛋白-2水平明显高于AGA、组,差异有统计学意义（P〈0.01）;脐血中胰岛素样生长因子-I与胰岛素水平呈正相关关系,而与胰岛素样生长因子结合蛋白-2呈负相关关系;脐血胰岛素、胰岛素样生长因子-I水平与新生儿出生体重呈正相关关系,而胰岛素样生长因子结合蛋白2与之呈负相关关系。结论脐血胰岛素、胰岛素样生长因子-I、胰岛素样生长因子结合蛋白2与胎儿生长受限的发生密切相关。     

人胰岛素样生长因子1基因转染对大鼠骨骼肌成肌细胞缺血再灌注损伤的影响

目的: 探讨人胰岛素样生长因子1(hIGF-1)基因转染对大鼠骨骼肌成肌细胞体外缺血再灌注损伤的影响及机制。方法: 分别用pLghIGF-1SN质粒(IGF组)和对照质粒pLgGFPSN(GFP 组)转染大鼠成肌细胞。未转染的成肌细胞(control组)作为细胞对照。转染后用免疫细胞化学、RT-PCR及ELISA检测基因表达。转染后3~14 d检测各组细胞的扩增率。转染的细胞接受缺血再灌注损伤后7 d,TUNEL法检测各组细胞的凋亡百分数,RT-PCR 检测各组细胞的bax和bcl-2 mRNA的表达,Western blotting检测各组细胞caspase-3的表达。结果: 基因转染后免疫细胞化学和RT-PCR 检测发现IGF组细胞有hIGF-1表达,GFP组和control组细胞未见hIGF-1表达;IGF组细胞上清中可检测到hIGF-1的蛋白表达,control组和GFP组细胞上清中未检测到hIGF-1蛋白表达。转染后14 d,IGF组细胞的扩增率显著大于control组和GFP组(P< 0.05);细胞经缺血再灌注损伤后,TUNEL染色检测结果显示:与GFP组相比,IGF组细胞的凋亡百分比显著降低(P< 0.05);RT-PCR 检测结果显示:与GFP组相比,IGF组细胞的bax mRNA表达显著降低,bcl-2 mRNA的表达显著增加(P< 0.05);Western blotting检测结果显示:与GFP组相比,IGF组细胞caspase-3蛋白表达显著降低。结论: IGF-1基因转染可增加成肌细胞的抗凋亡能力,其机制可能和降低Bax和caspase-3表达,增加Bcl-2的表达有关。     

腺病毒介导hIGF-1基因转染对软骨细胞增殖的影响

目的：探讨腺病毒介导人胰岛素样生长因子(human insulin—like growth factor，hIGF-1)基因转染对软骨细胞增殖的影响。方法：构建携带hIGF—1基因的重组腺病毒并进行PCR、Western blot鉴定。体外培养人胚胎软骨细胞，用处于对数增长期的第3代软骨细胞进行实验?分别转染1、10、100及500不同感染复数单位(multiplicity of infection，MOI)的Ad／hIGF-1，用PBS做阴性对照，hIGF-1生长因子(100μg/L)做阳性对照，采用四氮甲基唑蓝(methyl thiazolyl tetmzolium，MTT)法检测不同时间、不同组别软骨细胞吸光度。结果：第2代重组腺病毒上清液中PCR鉴定含有hIGF-1基因，Westem blot，证实Ad／hIGF-1表达成熟的hIGF-1生长因子。不同病毒滴度转染对软骨细胞增殖的影响存在量效依赖关系，100MO1软骨细胞吸光度约为对照组3倍，1MOI与10MOI、500MOI对软骨细胞增殖的影响近似；PBS组随着细胞体外培养时间延长，细胞增殖下降，hlGF-1生长因子、hIGF-1基因对软骨细胞增殖的影响存在时效关系，72h达到峰值。结论：Ad／hlGF-1基因转染对软骨细胞增殖的影响存在量效、时效依赖关系。     

转染hIGF-1基因增强兔退变椎间盘蛋白多糖的表达

目的 探讨人胰岛素样生长因子(hlGF-1)基因在退变椎间盘中的表达及对椎间盘中蛋白多糖(agglecan)的影响.方法 制备新西兰大白兔腰椎间盘退变(IDD)模型24只,随机分为Ad/CMV.hlGF-1、hlGF.1生长因子及PBS组,每组8只.IA-5、L5-6椎间盘中分别注射第2代Ad/CMV-hlGF-1(8×108PFU)、hlGF-1生长因子(100μg/L)、PBS均25μL.注射后1、2.4和8周,Western blot检测hlGF-1蛋白表达;RT-PCR检测aggrecan mRNA的表达.结果 hlGF-1蛋白带出现在7.6×103ku.Ad/CMV-hlGF-1组hIGF-I蛋白表达持续达4周以上,hIGF-1组表达持续约2周;PBS注射组无hIGF-1蛋白表达.aggrecan电泳条带出现在200～300 bp;在注射后1～4周,Ad/CMV-hlGF-I组内aggrecan mRNA相对表达量进行性增加,8周轻度下降,4个时期总的比较(F=8.51,P<0.05),注射后1～8周,hlGF-1组、PBS组aggrecan mRNA相对表达量进行性下降.结论 hlGF-1能够增强椎间盘aggrecan的表达.     

Erythropoietin promotes peripheral nerve regeneration in rats by upregulating expression of insulin-like growth factor-1

Introduction

Erythropoietin (EPO) has been shown to have beneficial effects on peripheral nerve damage, but its mechanism of action remains incompletely understood. In this study we hypothesized that EPO promotes peripheral nerve repair via neurotrophic factor upregulation.

Material and methods

Thirty adult male Wistar rats were employed to establish a sciatic nerve injury model. They were then randomly divided into two groups to be subjected to different treatment: 0.9% saline (group A) and 5000 U/kg EPO (group B). The walking behavior of rats was evaluated by footprint analysis, and the nerve regeneration was assessed by electron microscopy. The expression of insulin-like growth factor-1 (IGF-1) in the injured sciatic nerves was detected by immunohistochemical analysis.

Results

Compared to saline treatment, EPO treatment led to the growth of myelin sheath, the recovery of normal morphology of axons and Schwann cells, and higher density of myelinated nerve fibers. Erythropoietin treatment promoted the recovery of SFI in the injured sciatic nerves. In addition, EPO treatment led to increased IGF-1 expression in the injured sciatic nerves.

Conclusions

Erythropoietin may promote peripheral nerve repair in a rat model of sciatic nerve injury through the upregulation of IGF-1 expression. These findings reveal a novel mechanism underlying the neurotrophic effects of EPO.     

胰岛素样生长因子-1对雪旺细胞氧化损伤的保护作用

目的:探讨胰岛素样生长因子-1(IGF-1)对雪旺细胞氧化损伤的保护作用。方法:用体外纯化培养的雪旺细胞建立氧化损伤模型,将培养细胞分成氧化损伤组、IGF-1保护组和正常对照组。四甲基偶氮唑蓝比色法(MTT)检测细胞的活性,生化技术检测细胞内超氧化物歧化酶(SOD)的含量,免疫印迹法检测凋亡蛋白Bcl-2的表达水平。结果:与对照组相比,H2O2处理组细胞胞体积缩小、空泡化,细胞活性降低,SOD含量明显减少,Bcl-2表达减弱;而IGF-1保护组细胞存活率明显升高,SOD含量较损伤组高,Bcl-2表达明显上调。结论:IGF-1对氧化损伤的雪旺细胞有保护作用。     

Expression of insulin-like growth factor 1 in sarcomas

The expression of insulin-like growth factor-1 (IGF-1) was studied in normal tissues, in eight benign lesions and in 50 sarcomas. In palmar fibromatosis the spindle cells in cell-dense areas exhibited a strong immunoreactivity. IGF-1 was variably found in leiomyosarcomas (7/8), malignant schwannomas (7/9), synovial sarcomas (2/3), liposarcomas (3/6), fibrosarcomas (1/3), malignant fibrous histiocytomas (10/18) and in one angiosarcoma. Two rhabdomyosarcomas failed to express IGF-1 and only the spindle cell component of synovial sarcomas was positive. Immunoreactivity for IGF-1 in 10 malignant filrous histiocytomas (MFH) appeared to be related to co-expression of smooth muscle actin. These findings imply that MFHs can be subdivided into a group of tumours which are devoid of morphological and immunophenotypic evidence of differentiation and a group which manifest immunophenotypic differentiation.     

丝胶对Ⅱ型糖尿病大鼠海马生长激素/胰岛素样生长因子-1轴的作用

目的:探讨丝胶对Ⅱ型糖尿病大鼠海马生长激素(GH)/胰岛素样生长因子-1(IGF-1)轴的作用.方法:雄性SD大鼠随机分为正常对照组、糖尿病模型组、丝胶治疗组和阳性对照组.2％链脲佐菌素3d连续腹腔注射的方法建立Ⅱ型糖尿病大鼠模型后,丝胶治疗组和阳性对照组大鼠分别给予丝胶和二甲双胍灌胃治疗35 d.ELISA法检测各组大鼠血清GH和IGF-1水平,免疫印迹和RT-PCR法分别检测大鼠海马GH、生长激素受体(GHR)和IGF-1蛋白和mRNA的表达.结果:与正常对照组大鼠比较,糖尿病模型大鼠血清GH水平、海马GH的表达明显升高,血清IGF-1水平、海马GHR和IGF-1的表达明显降低;与糖尿病模型组大鼠比较,丝胶治疗组大鼠血清GH水平、海马GH的表达明显降低,血清IGF-1水平、海马GHR和IGF-1的表达明显升高.结论:丝胶可通过调节糖尿病海马GH/IGF-1轴的异常变化减轻海马损伤.     

Expression and distribution of insulin-like growth factor-1 receptor in human carcinomas

The insulin-like growth factor-1 receptor (IGF1-R) is a cellular receptor overexpressed in many tumor cell lines and in some human tumors that seems to play a critical role in transformation, tumorigenicity, and metastasis. To date, a comprehensive evaluation of tissue distribution of IGF1-R in human carcinomas from different anatomical sites has been lacking. Using stage-oriented human cancer tissue microarrays, we studied IGF1-R expression and distribution in a group of 152 human carcinomas from a variety of anatomical sites and from 63 normal tissues through immunohistochemistry. The tumors included carcinomas from breast (8), ovary (9), endometrium (7), esophagus (5), stomach (7), pancreas (7), liver (4), colon (10), kidney (14), bladder (17), prostate (11), head and neck (31), salivary glands (8), lung (13), and skin (1). Formalin-fixed, paraffin embedded tissues of each case were immuno-stained using the avidin-biotin peroxidase method and an anti-IGF1-R rabbit polyclonal antibody. High-membranous IGF1-R staining was observed in 7 of 8 (87.5%) breast carcinomas, in 9 of 9 (100%) ovarian carcinomas, in 7 of 7 (100%) endometrial carcinomas, in 5 of 7 (71.1%) gastric carcinomas, in 4 of 7 (57.1%) pancreatic carcinomas, in 9 of 10 (90%) colon adenocarcinomas, in 11 of 13 (84.6%) lung carcinomas, in 6 of 11 (54.5%) prostatic adenocarcinomas, and in 17 of 17 (100%) transitional cell carcinomas of the bladder. Only a minority of squamous cell carcinomas of the head and neck and esophagus (34), salivary gland tumors (5), and renal cell carcinomas (14) were IGF1-R positive. This study demonstrates the overexpression of IGF1-R across a wide variety of human carcinomas of glandular or transitional cell origin.     

间断性低氧暴露对红细胞参数及血清HIF-1α、EPO的影响

目的:探讨间断性低氧暴露及复氧休息对红细胞参数及血清低氧诱导因子-1α(HIF-1α)、促红细胞生成素(EPO)的影响.方法:制备间断性低氧动物模型,检测全血RBC、Hb、HCT红细胞参数,采用ELISA法检测血清HIF-1α、EPO水平,结合现场调查,检测间断性高原作业人员红细胞参数及EPO水平.结果:IH7、14、21、28 d组大鼠RBC、Hb、HCT水平明显高于常氧对照组(P<0.05),复氧后各参数水平下降.IH3、7、14 d组HIF-1α高于常氧对照组(P<0.05),EPO在IH3、7 d组高于对照组(P<0.05),复氧后HIF-1α、EPO水平下降.8个月组高原作业人员RBC水平高于平原对照组(P<0.05),Hb在2年组高于平原对照组(P<0.05).HCT与RBC大致呈同一规律,且2年组HCT仍明显高于平原对照组(P<0.05).与平原对照组比较,各组EPO的差异不显著.结论:间断性低氧暴露可以增加血清HIF-1α、EPO的含量,提高红细胞数量和血红蛋白的浓度,并随低氧周期的延长存在一定变化规律;复氧休息有利于低氧后机体的调整,使升高的红细胞参数及HIF-1、EPO水平下降.     

The association of insulin-like growth factor-1 standard deviation score and height in Chinese children with type 1 diabetes mellitus

Assessing the relationship between IGF-1 and height in type 1 diabetes children. Seventy-two type 1 diabetes children and 190 controls were recruited. The height standard deviation score of type 1 diabetes children was significantly higher than controls. The height standard deviation score was higher than the target height standard deviation score in both type 1 diabetes and controls. Serum IGF-1 levels and the IGF-1 standard deviation score were significantly lower in type 1 diabetes patients compared with controls. There was a significant difference in IGF-1 standard deviation score between the good glycemic control group and control group. The height standard deviation score was significantly correlated with C-peptide and IGF-1 levels. Furthermore, the IGF-1 standard deviation score was significantly correlated with glycemic control and C-peptide. The growth hormone/IGF-1 axis is impaired in type 1 diabetes, but height with good or poor glycemic control is not impaired.