首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到19条相似文献,搜索用时 203 毫秒
1.
原发性脑创伤后人脑皮层差异蛋白质组研究   总被引:2,自引:1,他引:1  
目的应用蛋白质组学技术,研究不同程度脑创伤后的人脑皮层蛋白质组表达变化的情况。方法将临床标本以GCS评分分为中度和重度损伤组,提取脑挫伤部位皮层的总蛋白。通过双向电泳.分离蛋白。应用胶内酶切、生物质谱,鉴定由图像分析软件所得出的具有表达差异的蛋白质点。结果通过比较,目前已发现12个蛋白质点,表达水平具有显著性差异变化。在已鉴定出的蛋白点中,属于10种蛋白质。依其功能可分为:细胞骨架、代谢反应、氧化应激反应、功能未知等几类。在重伤组中,一共有9个蛋白点、6种蛋白表达上调(α-烯醇化酶、磷酸丙糖异构酶、5’磷酸吡哆醇氧化酶、crtstalin、stathmin-1、NP25);中度损伤组3个蛋白点、4种蛋白表达上调(谷氨酰胺S转移酶、5’3’核苷酸酶、HSPC108、proapolipoprotein)。结论外伤后,神经系统由于受伤程度的不同,蛋白表达水平会发生变化。原发性脑创伤的损伤程度不同将产生神经系统病生理反应的差异。  相似文献   

2.
目的应用差异蛋白质组学技术,研究深低温和常温停循环后大鼠海马组织蛋白质的变化。方法采用深低温停循环模型,取大鼠海马组织,通过双向电泳、分离蛋白,然后通过胶内酶切、生物质谱分析差异的蛋白质点,鉴定出变化的蛋白质。结果通过Image Master软件分析报告发现有差异的Ratio值大于1.5的蛋白考染点14个。通过对这些蛋白考染点进行质谱鉴定,鉴定出28个蛋白质,其中4个为同一种蛋白,实际的蛋白数为24个。它们是细胞骨架蛋白、介导代谢的酶类、参与核酸合成、参与氧化应激反应的蛋白质及未知蛋白。结论鉴定出的差异蛋白质可能与深低温脑保护作用有关,某些蛋白质在低温脑保护中的作用尚未报道。  相似文献   

3.
目的对参与血-脑屏障组成的脑微血管内皮细胞在模拟缺血性脑损伤条件下的蛋白质组学变化特征进行研究.为深入理解缺血性脑损伤过程中血-脑屏障应答的分子学机制提供新的数据。方法采用比较蛋白质组学方法.对正常培养条件和氧糖剥夺培养条件下的小鼠脑微血管内皮细胞系(bEnd.3细胞)进行研究.分析二者之间的差异蛋白质表达谱。结果于氧糖剥夺培养条件下,小鼠脑微血管内皮细胞系中共发现52个蛋白质点的表达发生改变,通过串联飞行质谱成功鉴定45个蛋白质点,其中表达水平上调的41个.下调4个。根据基因本体论功能注释结果,这些蛋白质的功能分为糖代谢与能量产生、抗氧化损伤、细胞骨架重排、信号转导、可变剪接、蛋白折叠与降解等。其中约1/4的蛋白质与以往文献报道相似.即与缺血、缺氧损伤密切相关。结论这些差异蛋白质的发现可促进对血-脑屏障病理生理机制的了解.为深入理解缺血性脑损伤的分子学机制提供了新的数据,并将为后续研究提供参考。  相似文献   

4.
背景:热休克蛋白是生物体在不利环境因素刺激下应激合成的一种特殊蛋白质,热休克蛋白70参与移植免疫反应并发挥重要作用。 目的:探讨热休克蛋白70在恒河猴肝移植后急性排斥反应中的早期诊断价值。 方法:选用健康恒河猴采用改良二袖套+肝动脉重建的方法进行同种异体原位肝移植16例,移植后受体分为急性排斥组和对照组,每组8只,急性排斥组围手术期不行免疫抑制治疗,对照组围手术期使用免疫抑制治疗,分别在移植后6,12,24,72 h 4个时间点取移植后肝脏组织进行苏木精-伊红染色以判断排斥反应程度,western blotting检测肝组织中热休克蛋白70表达水平,免疫组化检测肝组织热休克蛋白70表达情况。 结论与结论:肝移植后72 h时段内急性排斥组肝脏急性排斥反应的组织学表现重于对照组,Baff分级水平高于对照组(P < 0.05),从移植后开始至移植后72 h两组移植肝经免疫组化和western blotting检测热休克蛋白70表达水平均有所升高,但急性排斥组要明显高于对照组(P < 0.05)。提示,未使用免疫抑制治疗的情况下,恒河猴肝移植后早期急性排斥反应在移植后72 h内即可明显观察到,热休克蛋白70在移植肝组织中表达水平也随着急性排斥反应的发生进展呈现明显上升趋势,对肝移植后早期急性排斥具有较高的预测和诊断价值。  相似文献   

5.
由多巴胺氧化产生的氧化应激和由此引发的蛋白质表达变化能够改变与帕金森病相关的细胞代谢过程。SH-SY5Y细胞经过10 μmol/L维甲酸和80 nmol/L 佛波酯相继诱导各72小时后出现了分化的神经元表现型,SH-SY5Y分化细胞暴露于100 μmol/L多巴胺24小时后出现氧化应激。在本研究中,为了在这个帕金森细胞模型中发现由氧化应激引发的蛋白质表达变化谱,我们通过二维差异胶内电泳和基质辅助激光解析电离飞行时间质谱的蛋白质组学方法,筛选和鉴定了显著性变化的蛋白质。结果显示,诱导分化作用使6个蛋白质表达显著性地变化(6个蛋白质表达均增加),毒性暴露作用使14个蛋白质表达显著性地变化(11个蛋白质表达增加,3个蛋白质表达降低);同时,在SH-SY5Y分化细胞中显著性地增加的微管蛋白alpha1,在SH-SY5Y分化细胞经过毒性暴露作用后显著性地降低,并且被鉴定为泛素化微管蛋白alpha。这提示,SH-SY5Y细胞在诱导分化作用后的蛋白质表达变化谱和SH-SY5Y分化细胞在毒性暴露作用后的蛋白质表达变化谱是不同但相关的。 关键词:SH-SY5Y细胞;维甲酸;佛波酯;多巴胺;氧化应激;蛋白质表达;蛋白质组学分析  相似文献   

6.
目的 比较放疗敏感性不同胶质瘤间蛋白质组差异,寻找可成为胶质瘤放射敏感性标记物的蛋白质.方法 收集放疗前的胶质瘤组织标本,以放疗后预后为判断标准,选择不同放疗敏感标本各3例,分为敏感组和耐受组.用二维液相色谱-质谱联用方法对肿瘤蛋白质组进行分析,筛选两组之问表达差异蛋白.应用Western印迹技术验证差异蛋白的表达.结果 丝切蛋白-1在放疗耐受组中表达明显增高,胶质瘤细胞大体形态无变化.结论 丝切蛋白-1可能与胶质瘤放疗敏感性相关.  相似文献   

7.
目的探讨急性脑梗死并发全身炎症反应综合征患者血清超敏C反应蛋白水平的变化及其对病情变化的意义。方法分别在发病24h内和第4、7天测定112例急性脑梗死患者血清超敏C反应蛋白水平的含量。结果 112例急性脑梗死患者中并发全身炎症反应综合征的患者共31例,发生率27.68%。在入院第4天超敏C反应蛋白水平比较,全身炎症反应综合征组与非全身炎症反应综合征组比较差异有统计学意义(P<0.05),在入院第7天2组比较差异有统计学意义(P<0.01)。结论超敏C反应蛋白的变化对急性脑梗死诱发全身炎症反应综合征和MODS有一定的预警价值。  相似文献   

8.
目的观察黄连素联合阿托伐他汀对急性缺血性卒中患者血清超敏C-反应蛋白和脂肪细胞型脂肪酸结合蛋白表达水平的影响,评价其临床疗效。方法 55例急性缺血性卒中患者随机分为阿托伐他汀20 mg/d组、阿托伐他汀40 mg/d组和黄连素0.40 g(3次/d)+阿托伐他汀20 mg/d组(联合治疗组),治疗3个月后比较血清超敏C-反应蛋白和脂肪细胞型脂肪酸结合蛋白表达变化。结果治疗前后3组患者血清超敏C-反应蛋白和脂肪细胞型脂肪酸结合蛋白表达水平差异均有统计学意义(P=0.023,0.000);治疗后3个月时,3组患者血清超敏C-反应蛋白和脂肪细胞型脂肪酸结合蛋白表达水平不同程度下降,分别为(1.69±2.29)和(281.43±311.05)mg/L、(7.81±12.48)和(321.59±289.35)mg/L、(2.16±3.34)和(376.55±249.72)mg/L,但组间差异未达到统计学意义(均P0.05),而且治疗药物与观察时间点之间不存在交互作用(均P0.05)。结论黄连素联合阿托伐他汀降低急性缺血性卒中患者血清炎症反应标志物超敏C-反应蛋白和脂肪细胞型脂肪酸结合蛋白表达水平的作用与阿托伐他汀强化治疗(40 mg/d)相似。  相似文献   

9.
目的 分析大鼠闭合性脑损伤后海马中蛋白质表达的变化及特点。 方法 选成年雄性SD大鼠随机分为手术对照组及损伤后不同时间点组, 复制Marmarou 落体打击闭合性颅脑损伤模型,应用蛋白质芯片技术和双向凝胶电泳技术分析大鼠脑损伤后海马中蛋白质表达的改变。并选择2个背景清晰、重复性及分辨率较好、在各组蛋白表达差异明显并呈现时序性变化的点进行质谱分析。结果 (1)蛋白质芯片结果显示,WCX-2芯片在海马中共获得364个蛋白质峰;IMAC-Cu芯片共获得345个蛋白质峰。 (2) 经双向凝胶电泳分离共获得18帧图,图谱蛋白质点分离效果良好,图谱平均匹配率>80%。PDQuest软件对蛋白质图谱进行定量分析证实,相对分子质量为58.0×103的差异蛋白质为葡萄糖调节蛋白,在损伤后4h、8h和12h表达水平上调;相对分子质量为28.4×103的差异蛋白质为蛋白酶体α亚单位3型,损伤后4h、8h、12h、24h和48h表达水平均上调。 结论 脑损伤可引起海马中蛋白质表达谱发生变化。葡萄糖调节蛋白和蛋白酶体α亚单位3型可能与创伤后脑损伤的发生、发展密切相关。  相似文献   

10.
目的应用蛋白质组学技术研究脑脊液的癫痫相关蛋白,以便从蛋白质水平揭示癫痫发生发展机制及可能的诊断标志物。方法双向凝胶电泳和质谱技术分离和鉴定癫痫组(n=8)和对照组(n=8)脑脊液的差异表达蛋白。结果约68个蛋白质点表达水平在两组间存在明显差异,选择18个差异蛋白质点质谱鉴定。除4个假想蛋白和未知功能蛋白及得到7个已知功能蛋白,涉及免疫反应、运输蛋白、能量代谢酶蛋白、细胞骨架蛋白和热休克蛋白。结论大部分被鉴定的蛋白为首次报道,这些蛋白与癫痫的发生发展相关,可能成为癫痫脑脊液分子标志物。  相似文献   

11.
To investigate molecular mechanisms of human brain aging, brain proteins were isolated from postmortem human young and old brains and profiled by two-dimensional gel electrophoresis (2-DE). With the help of special software, five down-regulated protein spots in two-dimensional gel electrophoresis gels of old brains were found compared with young brains, four of which was identified as a protein similar to peroxiredoxin 2 (accession-numbered as gi | 13631440), two of stathmin (phosphoprotein p19) and apolipoprotein A-I precursor (apo-AI) by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Eight common proteins, whose expressions were not altered between young and old brains, were also identified. The possible relevance of changes was analyzed. This study shows that the contribution of proteomics could be valuable in experimental gerontology field.  相似文献   

12.
Differential protein expression between various pathological grades of glioma has been shown in studies of glioma proteomics. However, very little data is available regarding normal brain tissues and glioma differential protein expression, because normal human brain tissues are difficult to harvest. The present study selected samples from low-grade astrocytomas and peritumoral brain tissues to analyze differential protein expression by two-dimensional (2D) electrophoresis and mass spectrometry techniques. Results revealing 36 protein spots by 2D electrophoresis, including 23 spots revealing increased expression and 13 spots revealing decreased expression. However, 25 differential proteins were identified by mass spectrometry, including 16 proteins with increased expression and 9 with decreased expression. Western blot analysis confirmed the mass spectrometry results, i.e., heat shock protein 70 (HSP70) and human transthyretin (TTR) expressions were increased, but glial fibrillary acidic protein (GFAP) was decreased, in astrocytomas. The present study constructed a 2D electrophoresis pattern between low-grade astrocytomas in the human brain and peritumoral tissues. Results demonstrated that a majority of differential proteins, such as HSP70, TTR, and GFAP, participate in malignant progression of gliomas.  相似文献   

13.
The reinforcing effects and long-term consequences of cocaine self-administration have been associated with brain regions of the mesolimbic dopamine pathway, namely the nucleus accumbens (NAc). Studies of cocaine-induced biochemical adaptations in rodent models have advanced our knowledge; however, unbiased detailed assessments of intracellular alterations in the primate brain are scarce, yet essential, to develop a comprehensive understanding of cocaine addiction. To this end, two-dimensional difference in gel electrophoresis (2D-DIGE) was used to compare changes in cytosolic protein abundance in the NAc between rhesus monkeys self-administering cocaine and controls. Following image normalization, spots with significantly differential image intensities (P<0.05) were identified, excised, trypsin digested and analyzed by matrix-assisted laser-desorption ionization time-of-flight time-of-flight (MALDI-TOF-TOF). In total, 1098 spots were subjected to statistical analysis with 22 spots found to be differentially abundant of which 18 proteins were positively identified by mass spectrometry. In addition, approximately 1000 protein spots were constitutively expressed of which 21 proteins were positively identified by mass spectrometry. Increased levels of proteins in the cocaine-exposed monkeys include glial fibrillary acidic protein, syntaxin-binding protein 3, protein kinase C isoform, adenylate kinase isoenzyme 5 and mitochondrial-related proteins, whereas decreased levels of proteins included beta-soluble N-ethylmaleimide-sensitive factor attachment protein and neural and non-neural enolase. Using a complimentary proteomics approach, the differential expression of phosphorylated proteins in the cytosolic fraction of these subjects was examined. Two-dimensional gel electrophoresis (2DGE) was followed by gel staining with Pro-Q Diamond phosphoprotein gel stain, enabling differentiation of approximately 150 phosphoprotein spots between the groups. Following excision and trypsin digestions, MALDI-TOF-TOF was used to confirm the identity of 15 cocaine-altered phosphoproteins. Significant increased levels were detected for gamma-aminobutyric acid type A receptor-associated protein 1, 14-3-3 gamma-protein, glutathione S-transferase and brain-type aldolase, whereas significant decreases were observed for beta-actin, Rab GDP-dissociation inhibitor, guanine deaminase, peroxiredoxin 2 isoform b and several mitochondrial proteins. Results from these studies indicate coordinated dysregulation of proteins related to cell structure, signaling, metabolism and mitochondrial function. These data extend and compliment previous studies of cocaine-induced biochemical alterations in human postmortem brain tissue, using an animal model that closely recapitulates the human condition and provide new insight into the molecular basis of the disease and potential targets for pharmacotherapeutic intervention.  相似文献   

14.
Extrapyramidal symptoms (EPS) commonly occur as side effects of antipsychotic drugs (APDs) and are most likely to arise when the occupancy of dopamine D(2) receptors in the striatum by these drugs exceeds 80%. We aimed to characterize changes in the protein expression profile in the striatum of rats after chronic (4 week) supra-therapeutic (EPS-inducing) treatment with risperidone (RIS), an atypical antipsychotic drug. Administration of RIS (2.1 mg/kg/day, via subcutaneous osmotic minipumps) induced significant vacuous chewing movements and catalepsy in male Sprague-Dawley rats over a 28-day treatment period compared with a vehicle (VEH) control group (n=12) (Karl et al., unpublished observation). Using two-dimensional gel electrophoresis (2DE), total protein extracts from the rat brain striatum were separated and protein expression was analyzed by Phoretix 2D Expression and Image Beta V4.02 software followed by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). 2DE gels resolved up to 450 protein spots, presumably different proteins and/or their isoforms. There were 30 protein spots showing statistically significant different densities between the RIS- and VEH-treated groups. All 30 proteins were successfully identified by MALDI-TOF MS, 28 of these were divided into groups based on their known functions. These included metabolic, signaling, transport, protein metabolism, chaperone, DNA binding and cell cycle categories. We conclude that chronic risperidone treatment accompanied by an EPS-like behavioral phenotype results in alterations in the striatal protein profile possibly subsequent to blockade of dopaminergic systems. These results suggest that possible mechanisms involved in APD-induced EPS include metabolic dysfunction and oxidative stress.  相似文献   

15.
Morphine has been used as a potent analgesic, having a high propensity to induce tolerance and physical dependence following their repeated administration. Although the mechanisms that underlie the development of dependence on morphine remain unclear, previous studies suggested that phosphorylations of diverse types of cellular proteins are crucial determinants of the neuroadaptive mechanisms associated with morphine dependence. Thus, understanding global phosphorylation events induced by chronic morphine administration is essential for understanding the complex signaling mechanisms of morphine dependence. This study characterized the alteration of tyrosine phosphorylation of frontal cortical proteins in morphine-dependent rat brains using a proteomic approach. Dependence was produced by continuous intracerebroventricular (i.c.v.) infusion of morphine (26 nmol/microl/h) for 72 h via osmotic minipumps in rats. Phosphotyrosyl (p-Tyr) protein spots in brain frontal cortical regions were detected by two-dimensional electrophoresis (2-DE) and immunoblotting with anti-p-Tyr-specific antibodies. The protein spots showing significant changes in tyrosine phosphorylation were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Similar patterns of protein expression were detected by 2-DE gels in morphine-dependent and saline-treated control rat brains. However, phosphotyrosine 2-DE images of the frontal cortical proteins from saline-treated control and morphine-dependent rat brains were apparently different. The densities of most matched p-Tyr protein spots were increased in morphine-dependent rat brains compared with that of control samples. Additional p-Tyr protein spots were detected in 2-DE image of morphine-dependent rat brains. Fifty of p-Tyr protein spots, corresponding to 40 different proteins, were identified from 2-DE gels of morphine-dependent rat brains. The identified proteins include enzymes, cytoskeletal proteins, cell signaling molecules, and other proteins. In conclusion, the first available phosphotyrosine proteomic resources of morphine dependence were established using an animal model. The findings illustrate the potential of proteomics as an effective technique for studying phosphorylation events of morphine dependence in brains.  相似文献   

16.
目的:使用蛋白质组学的技术手段,鉴定大鼠肾上腺皮质细胞瘤细胞系PC12细胞内的细胞骨架蛋白。方法:提取PC12细胞的蛋白质,建立固相pH梯度双向电泳图谱,应用图像扫描仪及ImageMaster 2D Elite分析软件获得蛋白质点的数字化和匹配性信息,挑选匹配良好的高峰度蛋白点,进行基质辅助激光解吸/电离飞行时间质谱(MALDI- TOF-MS)分析,鉴定。结果:用二维电泳技术分离,并用MALDI-TOF-MS成功鉴定出5个PC12细胞的细胞骨架蛋白。结论:PC12细胞蛋白质组中部分细胞骨架蛋白胶图位点的建立,为今后探讨这一类蛋白在神经系统疾病中的作用奠定了基础,并提供了新的侯选治疗靶点。  相似文献   

17.
目的 测定帕金森病(PD)脑脊液中蛋白的变化,为进一步探索PD的生物标记物提供线索.方法 采用荧光差异凝胶电泳技术分离并筛选PD和正常对照者脑脊液中差异表达蛋白质,用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)或串联质谱技术进行鉴定并分析.结果 共发现20个明显的差异蛋白点,其中11个点在PD中上调,9个点下调.共鉴定出8个蛋白质,其中有3个未知蛋白,均表现为下调.蛋白MYPT1出现明显下调,载脂蛋白原、脂蛋白发生明显上调,载脂蛋白A-I的一个异构体发生上调,一个异构体发生下调.结论 MYPT1与突触功能有关,载脂蛋白原、脂蛋白、载脂蛋白A-I与胆固醇代谢有关,这些蛋白与PD发生有一定关联,有可能成为PD的生物标记物.  相似文献   

18.
目的 采用蛋白质组学方法研究散发性颅内动脉瘤和健康成人外周血清蛋白表达差异.方法 对散发性颅内动脉瘤和健康成人的血清样本进行双向凝胶电泳分离,通过对两种蛋白图谱的比较,确定差异表达的蛋白点;而后对差异点进行基质辅助激光解析电离飞行时间质谱分析和蛋白数据库信息检索;随机选取2种蛋白进行Western blot法验证蛋白质组学实验结果.结果 在颅内动脉瘤和健康成人中鉴定出14种差异表达蛋白,得到了11个肽质量指纹图谱(peptide mass fingerprinting,PMF),最终7种蛋白质得到确认.结合珠蛋白、C反应蛋白、B因子、淀粉样血清蛋白A和载脂蛋白E前体在动脉瘤组表达上调;载脂蛋白A-Ⅳ前体和间α胰蛋白酶抑制物H4重链前体表达下调.蛋白印迹实验结果与双向电泳结果一致.结论 成功鉴定出7个与颅内动脉瘤相关的蛋白,为进一步阐明颅内动脉瘤形成的病理生理学机制提供了新的思路.  相似文献   

19.
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号