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1.
目的 研究蛋白酶体抑制对体外培养的星形胶质细胞(AS)IL-6分泌的影响.方法 SD乳鼠皮层AS原代培养,并纯化鉴定;不同浓度(0.1,1,2.5,5 μM)的蛋白酶体抑制剂(lactacystin)对第二代AS进行短期(24 h)急性处理,另一批同期的AS应用较低浓度(0.5μM)的lactacystin长期(4周)慢性处理,应用RT-PCR及ELISA方法检测AS IL-6 mRNA的表达和培养基中IL-6蛋白的分泌水平.结果 纯化传代的皮层AS经GFAP免疫荧光鉴定,其阳性率可达99.45%;短期抑制组中,1,2.5,5μM蛋白酶体抑制剂可诱导AS IL-6 mRNA表达和蛋白分泌增加,与对照组比较差异有显著性(P<0.01);长期抑制组中IL-6 mRNA表达及蛋白分泌也较同期对照组增加,差异有统计学意义.结论 一定程度蛋白酶体活性抑制可诱导培养的AS IL-6 mRNA表达及蛋白分泌增加,提示蛋白酶体功能障碍后可能通过诱导AS分泌IL-6增加来参与阿尔茨海默病的病理改变. 相似文献
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目的 探讨细胞外信号调节激酶(ERK1/2)的磷酸化水平对体外星形胶质细胞(Ast)增殖及其细胞周期的影响.方法 将培养成熟的大鼠原代Ast分为两组,其中一组用ERK1/2磷酸化的特异性抑制剂U0126进行处理(U0126组),另一组不做处理,作为对照组.通过Western Blot技术、Click-iT Edu技术和流式细胞术分别检测两组Ast中ERK1/2磷酸化水平、Ast增殖细胞百分比及各期细胞百分比.结果 U0126组ERK1/2的磷酸化水平(39.13%±6.71%)明显低于对照组(100%).U0126处理24 h后,U0126组Ast增殖细胞百分比[(2.63±1.14)%]低于对照组[(21.43±3.81)%](t=21.13,P<0.01).G1期U0126组Ast[(93.67±0.68)%]高于对照组[(84.63±1.00)%](t=12.91,P<0.01);S期U0126组Ast[(2.90±0.23)%]低于对照组[(14.21±1.14)%] (t=16.87,P<0.01);G2期U0126组Ast[(3.43±0.88)%]高于对照组[(2.08±0.21%)%](t=4.35,P<0.05).结论 降低ERK1/2的磷酸化水平可抑制Ast的增殖,并将其阻断在G1期,抑制Ast从G1期向S期的转化,同时抑制其分裂将其阻断在G2期. 相似文献
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周期素依赖性激酶抑制剂(cyclin-dependent kinases inhibitors,CDKIs)分为INK4(inhibitor of CDK4,INK4)和KIP(kinase inhibition protein,KIP)两大家庭,主要通过视网膜母细胞瘤蛋白(retinoblastoma protein,pRb)或p53两条途径对细胞周期起调控作用。在脑缺血再灌注损伤后周期素依赖性激酶(cyclin-dependent kinases,cDKs)所调控的信号通路被激活,细胞周期紊乱可促使神经元凋亡,CDKIs对神经元损伤具有保护作用。 相似文献
4.
目的比较研究成年大鼠细胞周期蛋白依赖性激酶抑制因子在神经元和星形胶质细胞的表达差异。方法应用免疫荧光和激光扫描共聚焦显微镜观察成年大鼠生理状态下大脑皮层或海马CA1、CA3、DG区神经元和星形胶质细胞细胞周期蛋白依赖性激酶抑制因子(CDKI)p15Ink4b、p21cipl的表达。结果成年大鼠海马区和大脑皮层的神经元有p15Ink4b和p21cipl的表达,细胞核和细胞浆均有表达,且以胞核为主;星形胶质细胞也有上述细胞周期调控蛋白的表达,便细胞数目较少,并且表达这些指标的星形胶质细胞多聚集在海马区。结论成年大鼠大脑皮层和海马区的神经元和星形胶质细胞均表达p15Ink4b和p21cipl,而其在神经元的表达较星形胶质细胞更为普遍。 相似文献
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目的 探讨人脑胶质瘤中脾酪氨酸激酶(Syk)的表达及其与周期索1(CyclinDl)水平的相关性.方法 收集贵州航天医院神经外科自2005年1月至2010年1月间手术切除并经病理证实的脑胶质瘤标本46例,其中Ⅰ级13例,Ⅱ级9例,Ⅲ级5例,Ⅳ级胶质母细胞瘤19例.另取5例因脑创伤行内减压术患者的正常脑组织标本作为对照,RT-PCR检测脑组织标本Syk mRNA、CyclinD1 mRNA的表达并分析二者的相关性.结果 与正常脑组织比较,Ⅰ、Ⅱ、Ⅲ、Ⅳ级脑胶质细胞瘤标本Syk mRNA表达较低,CyclinDl mRNA表达较高,差异有统计学意义(P<0.05),而且胶质瘤的病理级别越高,Syk mRNA的表达越低,CyclinDl mRNA表达较高,差异均有统计学意义(P<0.05);胶质瘤中Syk与CyclinDl的表达呈负相关关系(r=-0.832,P=0.000).结论 Syk在脑胶质瘤中表达较低或缺失,提示其可能具有抑癌基因功能,其机制可能与下调胶质瘤中CyclinD1的表达有关. 相似文献
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目的 观察沉默细胞周期蛋白依赖性激酶5(Cdk5)对星形胶质细胞活化增殖和细胞周期的影响。方法 培养及鉴定SD大鼠星形胶质细胞,设计合成针对星形胶质细胞Cdk5的小干扰RNA(siRNA),将Cdk5 siRNA转染入星形胶质细胞,通过Real-time PCR和Western blot检测沉默效率; 分为对照组及Cdk5 siRNA干预组,在不同时间点(3、6、12及24 h)采用Edu染色及流式细胞术检测细胞增殖及细胞周期情况。结果 成功利用Cdk5 siRNA沉默星形胶质细胞Cdk5; Edu染色显示Cdk5 siRNA干预组干预3、6、12 h Edu染色阳性率较对照组显著降低(P<0.01); 流式细胞术显示 Cdk5 siRNA干预组干预3、6、12 h处于S期的星形胶质细胞比例较对照组显著降低(P<0.05)。结论 沉默Cdk5可抑制星形胶质细胞的增殖及细胞周期进展,提示Cdk5在星形胶质细胞增殖过程中起到重要作用。 相似文献
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目的 分析不同放疗敏感性星形胶质细胞瘤蛋白表达特点,筛选与放射耐受关系密切的蛋白质.方法 通过随访胶质瘤切除术后接受三维适型调强放射治疗的患者,按其存活状况及生活质量分为放疗敏感组(7例)和耐受组(8例),病理诊断均为星形胶质细胞瘤(WHOⅡ级).采用二维液相色谱串联质谱法(2D-LC-MS/MS)对入组肿瘤标本进行蛋白组学分析,筛选差异表达蛋白.并对放疗耐受相关蛋白丝切蛋白-1(cofilin-1)及磷酸甘油酸激酶1(phosphoglycerate kinase 1,PGK1)用免疫印记法(Western blot)检测其表达量.结果 2D-LC-MS/MS共鉴定出两组间差异表达蛋白36个,以cofilin-1与PGK1含量最为丰富,在放疗耐受组肿瘤中呈高表达状态.Western blot 检测结果与蛋白组学分析相吻合.结论 放疗后预后不同的同级别星形胶质细胞瘤间蛋白表达差异明显,以cofilin-1及PGK1为代表,可能成为星形胶质细胞瘤放疗耐受的潜在生物标志物. 相似文献
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目的 探讨缺氧/复氧对于体外培养星形胶质细胞表达AQP4的影响及缺血性脑血管病脑水肿的发病机制.方法 选取新生24 h Wistar大鼠脑组织行星形胶质细胞原代培养并建立缺氧/复氧模型,应用Western blot和免疫细胞化学法检测星形胶质细胞AQP4的表达变化.结果 与对照组比较,缺氧3 h和6 h星形胶质细胞AQP4蛋白表达减少(P<0.01),复氧后6 h和9 h其表达明显增高(P<0.01).结论 AQP4表达变化与细胞损害相关. 相似文献
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目的探讨丹参酮ⅡA对体外培养星形胶质细胞细胞周期和增殖的影响。方法本研究采用体外细胞培养方法获得新生SD小鼠大脑皮质星形胶质细胞,然后在培养液中分别加入不同浓度的丹参酮ⅡA,作用8 h及24 h后,通过流式细胞仪检测丹参酮ⅡA对星形胶质细胞细胞周期的影响。结果丹参酮ⅡA作用于星形胶质细胞后,可引起细胞周期的显著变化,表现为G0/G1期细胞数减少,S期、G2/M期细胞数增多。结论丹参酮ⅡA可促进体外培养的星形胶质细胞增殖。 相似文献
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目的:探讨人脑胶质瘤中βcatenin的表达及与细胞增殖的相关性。方法:用免疫组化法观察了47例胶质瘤标本βcatenin、cyclinD1、增殖细胞核抗原(PCNA)的表达,并分析其相关性。结果:在胶质瘤中βcatenin、cyclinD1、PCNA均随肿瘤恶性程度增高而增高(P分别为0.017、0.017、0.027),相关分析发现βcatenin、cyclinD1、PCNA的表达两两相关(P<0.001,P<0.001和P=0.005)。结论:βcatenin表达水平的异常与胶质瘤细胞周期调节紊乱、增殖指数上升有关,因此可能是控制肿瘤细胞增殖活性的候选基因之一。 相似文献
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Cyclin D1 in astrocytic tumours: an immunohistochemical study 总被引:1,自引:0,他引:1
Forty-eight astrocytic tumours were stained immunohistochemically with antibodies to the cell cycle-regulating protein, cyclin D1, and to the proliferation marker MIB1 (Ki-67) using formalin fixed paraffin embedded tissue and a microwave antigen retrieval system. Cases were classified by the WHO system (1993). The labelling indices (LI) for both antibodies were compared with each other and with the tumour type. The mean labelling indices for both antibodies increased with the degree of malignancy, and a significant difference was seen between the pilocytic astrocytoma and diffuse astrocytoma together vs anaplastic astrocytoma and glioblastoma together. However, within each tumour type there was considerable variation in the labelling indices and a clear cut off value could not be demonstrated. There was a strong positive correlation between labelling indices for cyclin D1 and MIB1 in diffuse astrocytoma, but this correlation broke down increasingly in anaplastic astrocytoma and glioblastoma. There was poor correlation between cyclin D1 and MIB1 in pilocytic astrocytoma, a feature which appeared to separate them from the diffuse astrocytoma. Average labelling indices for cyclin D1 were higher than those of MIB1, which suggests that cyclin D1 positive cells represent a pool of cells from which proliferation and hence MIB1 expression can take place. In conclusion, cyclin D1 is overexpressed in astrocytic tumours, more so with increasing grade of malignancy and in a way which approximately correlates with MIB1 expression. 相似文献
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目的 探讨cyclin D1/D2/D3在大鼠脑胶质瘤组织中的表达.方法 将体外培养的大鼠C6胶质瘤细胞(细胞数为3×105个)借助动物立体定向仪接种于Wistar大鼠左侧尾状核区,解剖标本,行组织病理学检查及GFAP、cyclin D1/D2/D3免疫组化检测.结果 cyclinD1/D2/D3均呈阳性表达.cyclin D1/D3主要在细胞胞核表达,cyclin D2主要在胞浆表达,部分在胞核表达,不同存活期荷瘤鼠瘤组织中cyclin D1/D2/D3表达量不同.结论 cyclinD1/D2/D3过分表达参与调节大鼠脑胶质瘤增殖.荷瘤鼠生存期与cyclin D家族蛋白的表达呈负相关关系. 相似文献
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目的观察脑缺血后细胞周期蛋白(cyclin)D1和它的酶CDK4基因表达,以及这种表达的改变是否影响神经细胞凋亡。方法成年雄性SD大鼠32只随机分为假手术组(n=4)和实验组(n=28),实验组再进一步分为7个亚组(再灌注2h,6h,12h,1d,3d,7d和14d,每组n=4)。应用线栓法建立SD大鼠大脑中动脉阻塞/再灌注模型.TUNEL法检测神经细胞凋亡。原位杂交检测cyclinD1和CDK4mRNA的表达。结果CyclinD1mRNA和CDK4mRNA的表达与凋亡细胞的区域基本相同。再灌注2h脑组织即开始出现神经细胞凋亡,并于1d分别在皮层区和纹状体区达高峰(分别为72.80±4.66和87.75±0.85)。神经细胞cyclinD1mRNA和CDK4mRNA的表达分别于再灌注2h和6h开始逐渐增强,并于12h和1d达高峰(皮质区分别为94.50±2.75和85.75±3.73,纹状体区分别为88.25±5.06和89.80±2.93)。结论CyclinD1和CDK4选择性地在形态学完整或已经有改变的缺血侧神经元和少突胶质细胞内表达。CyclinD1/CD1(4mRNA表达可能是诱导细胞凋亡的重要因素之一。 相似文献
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Ivana Bosone Paola Cavalla Loredana Chiad‐Piat Nicoletta Di Vito Davide Schiffer 《Neuropathology》2001,21(3):155-161
Cyclin D1 regulates G1–S progression. In many carcinomas it is overexpressed and it might even correlate with prognosis. However, the amplification of CCND1 contributes to the loss of cell cycle control only in a small fraction of malignant gliomas. Cyclin D1 can be immunohistochemically demonstrated by DCS‐6 mAb. In astrocytic gliomas the fraction of tumor cells with positive nuclei is almost null in well differentiated tumors and increases with the increase of proliferation rate that occurs in anaplasia. The correct evaluation of this fraction is hindered by the positive staining of normal oligodendrocytes and microglia cells. The cyclin D1‐positive staining of normal oligodendrocytes and microglia cells has been studied in a series of 20 oligodendrogliomas, five diffuse astrocytomas and five oligoastrocytomas and in 10 samples of normal cortex and white matter, using cyclin D1 DCS‐6 mAb, Feulgen reaction and CR3.43 mAb for microglia cells. As well as microglial nuclei, the nuclei of normal oligodendrocytes of the cortex and white matter, including peri‐neuronal satellites and pericapillary cells, were immunostained by DCS‐6 mAb. In infiltrative areas of oligodendrogliomas, normal, cyclin D1‐positive oligodendrocytes and cyclin D1‐negative tumor cells coexisted. In anaplastic oligodendrogliomas, cycling tumor oligodendrocytes may regain the capacity to express cyclin D1, which is thus positive in some tumor cells. The occurrence of positive oligodendrocytes in the peripheral parts of tumors can be useful in distinguishing astrocytomas from oligoastrocytomas. 相似文献
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血红素加氧酶在阿尔茨海默病中的表达及意义 总被引:1,自引:0,他引:1
目的:研究血红素加氧酶(HO-1)在阿尔茨海默病(AD)额叶和海马中的表达,探讨其神经保护作用及机制.方法:选取诊断明确的AD尸检脑标本14例(AD组);另选取中枢神经系统以外疾病、无显著脑病理改变的死亡病例32例为对照组.采用免疫组织化学染色法,以阳性表达程度和积分吸光度(IA)值观察HO-1在AD及对照组额叶和海马中的表达;采用免疫荧光双重染色方法,观察AD脑中HO-1与GFAP、Tau的共同表达.结果:AD组额叶与海马HO-1阳性率较对照组增高,且差异有统计学意义( P〈0.05);对照组不同年龄段HO-1阳性率随年龄增高,青年组与高龄组比阳性率差异有统计学意义(P〈0.05);AD组IA值较对照组增高( P〈0.05),AD组海马IA值较额叶增高且差异有统计学意义( P〈0.05);部分神经元胞质内HO-1与Tau共同表达,星形胶质细胞内HO-1与GFAT共同表达.结论:HO-1可能通过启动内源性神经保护机制在AD中起脑保护作用. 相似文献
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Effect of growth factors and steroid hormones on heme oxygenase and cyclin D1 expression in primary astroglial cell cultures 下载免费PDF全文
V. Bramanti S. Grasso D. Tomassoni E. Traini G. Raciti M. Viola G. Li Volti A. Campisi F. Amenta R. Avola 《Journal of neuroscience research》2015,93(3):521-529
Astrocyte activity may be modulated by steroid hormones and GFs. This study investigates the interaction between glucocorticoids or estrogens and GFs on the expression of heme oxygenase‐1 (HO‐1) and cyclin D1 in astrocyte cultures at 14 days treated for 48 or 60 hr with dexamethasone (DEX) or 48 hr with 17β‐estradiol (E2) alone or with GFs added only in the last 12 or 24 hr. Twelve‐ or twenty‐four‐hour epidermal growth factor (EGF) treatment significantly enhanced HO‐1 expression in astrocyte cultures pretreated for 48 hr with DEX. A highly significant increase in HO‐1 expression was obtained after the last‐12‐hr EGF treatment in 48‐hr E2‐pretreated astrocyte cultures; this enhancement was particularly significant in 48‐hr E2‐pretreated cultures as well as in the last‐12‐hr insulin‐treated ones pretreated for 48 hr with E2. Sixty‐hour DEX‐alone pretreatment as well as the last‐12‐hr EGF treatment in 60‐hr DEX‐pretreated astrocyte cultures showed a significant increase of cyclin D1 expression. A significant decrease of cyclin D1 expression in the last‐12‐hr insulin‐like growth factor‐I (IGF‐1)‐treated cultures pretreated for 60 hr with DEX was observed. A highly significant enhancement in cyclin D1 expression in 14 days in vitro astrocyte cultures pretreated with E2 alone for 48 hr and treated in the last 12 hr with IGF‐1 in 48‐hr E2‐pretreated cultures was found. Finally, the data highlight an interactive dialogue between the growth factors and glucocorticoids or estrogens during the maturation of astroglial cells in culture that may control the HO‐1 and cyclin D1 expression as well as proliferating astroglial cells during the cell cycle. © 2014 Wiley Periodicals, Inc. 相似文献
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目的探究星形胶质细胞内PTEN诱导假定激酶1(PINK1)缺失对缺血时神经保护作用的影响及其作用机制。方法离体培养原代星形胶质细胞,使用小干扰RNA(si RNA)沉默PINK1表达,氧糖剥夺(OGD)建立细胞缺氧模型,分为4组:PINK1沉默组(si RNA+转染剂)、空质粒组(空质粒+转染剂)、转染剂组(只加转染剂)和对照组(星形胶质细胞),各组均与神经元共培养;另设立神经元单独培养组。免疫荧光染色观察神经元凋亡情况。定量PCR及ELISA检测星形胶质细胞促红细胞生成素(EPO)及血管内皮生长因子(VEGF)表达量;Western blot检测星形胶质细胞内缺血诱导因子(HIF)及核因子κB(NF-κB)通路相关蛋白水平。结果 OGD损伤后神经元凋亡率较高,与星形胶质细胞共培养后神经元凋亡率显著降低(P0.05)。PINK1基因沉默后共培养神经元凋亡增加,星形胶质细胞EPO及VEGF分泌量减少、胞内EPO及VEGF转录水平降低(P0.05);HIF-1、HIF-2与NF-κB通路活化水平均显著降低(P0.05)。结论星形胶质细胞对OGD损伤神经元有保护作用,其作用通过EPO及VEGF实现;PINK1基因沉默后星形胶质细胞对缺血神经元保护作用减弱,可能与NF-κB通路活化水平降低、HIF激活受损进而下调EPO和VEGF表达量有关。 相似文献
19.
胶质瘤PTEN和细胞周期蛋白D1表达及其意义 总被引:6,自引:5,他引:1
目的 探讨胶质瘤组织中PTEN和细胞周期蛋白D1(CyclinD1)表达的意义。 方法 应用免疫组化结合计算机图像分析方法检测PTEN和CyclinD1蛋白在 80例胶质瘤的表达及其相关关系。结果 PTEN阳性率为 6 3.8% (5 1/ 80 ) ,低分级胶质瘤 (I~II)阳性率显著高于III级和IV级 (P<0 .0 1) ;胶质瘤不同病理分级间CyclinD1蛋白表达存在显著性差异 (P <0 .0 1)。胶质瘤细胞CyclinD1与PTEN蛋白表达呈显著性负相关关系 (r=- 0 .5 4 3,P <0 .0 1)。结论 PTEN蛋白表达可能在胶质瘤细胞周期调节和细胞增殖中发挥重要作用 相似文献