水通道蛋白在正常大鼠椎间盘组织中的表达与分布

目的：阐明水通道蛋白在椎间盘中的表达类型及分布情况。方法：收集正常大鼠椎间盘组织，部分用于提取RNA，采用RT-PCR的方法检测水通道蛋白表达的类型；部分标本固定后行组织切片，进行AQPs(aquaporin)的免疫组织化学染色，观察其表达与分布。结果：在正常大鼠的椎间盘组织中软骨细胞、纤维环细胞和髓核细胞皆有AQP1、3的表达，软骨终板中心地带的表达强于周边区域，内层纤维环表达强于外层纤维环；其余几种水通道蛋白未见表达。结论：AQPl、3在正常大鼠椎间盘中的表达及其空间分布提示其可能与椎间盘内水、甘油的代谢有关，对维持椎间盘组织的正常功能可能有重要的作用?     

水通道蛋白1和3在小鼠前列腺组织的表达及意义

目的:揭示水通道蛋白(AQP)在小鼠前列腺组织的表达及意义,为深入研究前列腺正常及某些病理状态下AQPs表达调控及生理功能奠定基础。方法:取小鼠前列腺组织,采用RT-PCR方法检测AQP 0~4mRNA在前列腺组织的表达,同时应用Western印迹和免疫组化研究AQP1及AQP3在前列腺组织的定位表达。结果:RT-PCR显示前列腺组织中有AQP1、3 mRNA表达,而未见AQP0、2、4 mRNA的表达。Western印迹结果显示在正常前列腺组织中表达相对分子质量为28 000的AQP1蛋白,以及相对分子质量分别为35 000和27 000的糖基化和非糖基化AQP3蛋白。免疫荧光和免疫组化结果表明小鼠前列腺近管腔处分泌细胞的胞内囊泡及细胞膜均可见较强的AQP1蛋白表达;而在前列腺基质上皮细胞有AQP3的表达。结论:AQP1和AQP3基因在前列腺分泌上皮细胞表达,提示AQP1和AQP3可能通过促进前列腺上皮细胞对水的渗透,在前列腺液分泌过程中发挥重要的生理作用。     

水通道蛋白1和3在结直肠癌组织中的表达及其意义

目的 探讨水通道蛋白1（AQP-1）和AQP-3与结直肠癌发生发展的关系.方法 应用免疫组织化学染色SP法检测25例结直肠腺瘤、50例结直肠癌及相应癌旁组织中AQP-1和AQP-3的表达.结果 结直肠癌组织中AQP-1的阳性表达率为64%（32/50）,明显高于癌旁组织（38%,19/50）和腺瘤组织（32%,8/25）,差异有统计学意义（P＜0.01）;AQP-1表达与肿瘤浸润深度和淋巴结转移有关（P＜0.05）.AQP-3在结直肠癌、腺瘤及癌旁组织中的阳性表达率分别为52%（26/50）、44%（11/25）和56%（28/50）,差异无统计学意义（P＞0.05）;AQP-3表达与患者年龄、肿瘤大小及浸润深度有关（P＜0.05）.结直肠癌中AQP-1与AQP-3表达无显著相关性（P＞0.05）.结论 结直肠癌中高表达AQP-1,但AQP-3表达却未见增高;AQP-1和AQP-3表达无相关性.     

水通道蛋白1和3在结直肠癌组织中的表达及其意义

Objective To investigate the significance of aquaporin-1 (AQP-1) and aquaporin-3 (AQP -3) in the development of colorectal carcinoma. Methods The expression of AQP-1 and AQP-3 was investigated using immunohistochemical staining with Streptavidin Perdcidase in tissues from colorectal adenoma (CRA, n=25), colorectal cancer (CRC, n=50), and adjacent mucosa (CRT, n=50).Results The positive rate of AQP-1 was 64%(32/50) in CRC, significantly higher than that in CRT (38%, 19/50) and CRA(32%,8/25)(P＜0.05). The expression of AQP-1 was associated with depth of invasion and lymph node metastasis in CRC patients(P＜0.05). The positive rate of AQP-3 was 56% in CRT, 44% in CRA, and 52% in CRC. There were no significant differences(P＞0.05). The expression of AQP-3 was associated with age, tumor diameter, and depth of invasion(P＜0.05). No significant correlation between the expression of AQP-1 and AQP-3 in CRC was shown by Spearman correlation analysis(P＞0.05). Conclusions AQP-1 expression is increased in CRC while the expression of AQP-3 is not. There is no correlation between the expression of AQP-1 and AQP-3 in CRC.     

椎间盘退变时水通道蛋白1表达变化研究

目的阐明水通道蛋白1在椎间盘退变时表达变化特点。方法采用腰椎失稳的方法建立大鼠腰椎间盘退变的动物模型,影像学观察模型建立的可靠性,收集大鼠椎间盘组织,部分用于提取RNA,采用RT-PCR的方法检测水通道蛋白1mRNA表达的变化,部分标本用western blot进行AQP1蛋白的检测。结果腰椎失稳可以建立大鼠腰椎间盘退变模型,AQP1mRNA的表达随椎间盘退变的进程进行性下降,其蛋白表达也呈现进行性下降的趋势,术后3月时同术前相比相差显著(P<0.05),术后6月和9月则下降更为明显(P<0.01);而在手术对照组而言,于术后6月时也出现AQP1基因和蛋白表达水平的下降(P<0.05)。结论腰椎间盘退变时,AQP1在基因和蛋白的表达水平下降,且与退变的时间长短相关,提示AQP1参予了椎间盘退变的发生发展,AQP1表达的下降可能是引起椎间盘退变的原因之一。     

水通道蛋白4在食管鳞癌中的表达

目的 观察水通道蛋白4(AQP4)在食管鳞癌组织中的表达并探讨其在食管癌发病中的作用.方法 取食管鳞癌组织、癌旁正常鳞状上皮组织各16例,应用免疫组织化学,逆转录.聚合酶链反应(RT-PCR)技术检测AQP4的表达及分布.结果 免疫组织化学显示,AQP4表达于正常食管黏膜鳞状上皮细胞,在食管鳞癌组中主要表达于肿瘤上皮细胞和癌巢中.RT-PCR法结果 显示,AQP4在癌旁正常组织和食管癌组织中的mRNA表达平均相对A值分别为0.45±0.12、0.70±0.23,差异有统计学意义(P＜0.05).结论 AQP4在正常食管黏膜鳞状上皮细胞以及食管鳞癌组中均有表达,而且在癌组织中表达增加;AQP4可能对人食管癌的发生、发展起促进作用.     

不同状态小肠水通道蛋白3的表达分析及其临床意义

目的 研究梗阻小肠组织中水通道蛋白3(AQP3)的表达并分析其意义。方法 对25例正常小肠、25例充血水肿小肠组织和坏死小肠组织行病理切片和苏木精-伊红染色、AQP3免疫组织化学染色,观察结构变化及AQP3表达情况。结果 正常小肠AQP3呈强阳性表达,随着小肠梗阻病程的进展,小肠充血水肿,纤维组织增生及血管扩张,AQP3的表达随之降低,差异有统计学意义（P<0.001）。进展至坏死阶段,小肠组织结构的破坏,AQP3呈零表达,AQP3表达降低的小肠梗阻患者炎性反应更严重（P<0.05）。结论 自正常小肠至坏死小肠,AQP3的表达呈阶梯式下降,提示AQP3可能是预示梗阻小肠坏死的可靠标志物。     

水通道蛋白1在不同程度退变腰椎间盘中的表达及意义

目的 观察水通道蛋白1在退变腰椎间盘组织中的表达变化,研究其与椎间盘退变的关系。方法 实验组取材于经手术治疗腰椎间盘突出症患者椎间盘标本30例,突出型、脱出型、游离型各10例;对照组取材于腰椎损伤致椎体骨折患者切除椎间盘10例。应用免疫组织化学法检测水通道蛋白1在椎间盘组织中的表达。结果 （1）苏木精-伊红染色显示,对照组椎间盘组织仍保持着致密的板层结构,实验组椎间盘组织多呈纤维化样改变。（2）各组髓核细胞中水通道蛋白1表达均明显高于同组纤维环细胞中的表达(P≤0.01)。（3）实验组突出型椎间盘标本髓核及纤维环细胞中水通道蛋白1表达明显低于对照组(P＜0.01),脱出型和游离型椎间盘标本中水通道蛋白1表达明显低于突出型 (P＜0.01)。结论 水通道蛋白1随着椎间盘突出、脱出、游离不同程度退变,表达呈逐渐下降趋势。     

兔椎间盘终板细胞凋亡与水通道蛋白-1的关系

目的 探讨水通道蛋白(AQP)-1与椎间盘终板细胞凋亡的关系及意义.方法 将23个含终板的完整椎间盘分为4组:A组直接把取出的新鲜完整的椎间盘作为对照(椎间盘×5),B组为无干预组[进行培养但不加白细胞介素(IL)-1β,椎间盘×6],C组加入10μg/L IL-1β(椎间盘数×6),D组加入30μg/L IL-1β(椎间盘数×6).B、C、D组培养2周后取组织的相邻切片,免疫组织化学检测AQP-1和Caspase-3的表达.结果 免疫组织化学显示A组中AQP-1和Caspase-3阳性细胞率分别为(88.4±1.1)%和(5.5±0.8)%,B组阳性细胞率分别为(86.7±1.4)%和(6.3±1.6)%,C组中阳性细胞率分别为(42.8±2.2)%和(62.0±1.7)%,D组阳性细胞率分别为(30.5±2.0)%和(71.8±1.23)%,AQP-1与Caspase-3表达呈负相关.结论 AQP-1表达减少与终板细胞的凋亡密切相关.     

功能性便秘患者结肠黏膜水通道蛋白3和水通道蛋白9的表达及意义

目的研究功能性便秘患者结肠黏膜水通道蛋白3（AQP3）和水通道蛋白9（AQP9）的表达与分布情况．并探讨其与便秘的关系。方法采用免疫组化技术和Western印迹半定量检测法检测45例功能性便秘患者（试验组）和21例无便秘患者（对照组）结肠黏膜AQP3和AQP9，把AQP3和AQP9与相应内参蛋白β-actin的灰度值之比视为AQP3和AQP9的相对含量。结果免疫组化检测显示：AQP3主要表达在结肠黏膜顶部的吸收细胞．且主要分布于细胞膜的基底侧和腔面，杯状细胞少见阳性染色，AQP9主要表达在黏膜腔面的杯状细胞，且主要分布于细胞膜的基底侧。Western印迹检测结果：试验组和对照组升结肠AQP3灰度比平均值分别为0．905和0．798（P〈0．05）。AQP9灰度比平均值分别为0．544和0．543（P〉0．05）；试验组和对照组降结肠AQP3灰度比平均值分别为0．697和0．701（P〉0．05），AQP9灰度比平均值分别为0．575和0．732（P〈0．05）。结论功能性便秘患者升结肠黏膜AQP3表达高于非便秘者，而其降结肠AQP9表达则低于非便秘者；AQP3和AQP9含量的增减可能在便秘的发生发展过程中发挥了重要作用。     

白细胞介素1对椎间盘细胞软骨特异性基因Sox9 mRNA的调节作用

目的探讨白细胞介素1（interleukin-1,IL-1）对人椎间盘细胞软骨特异性基因Sox9和Ⅱ型胶原基因表达的调节作用。方法应用RT—PCR技术检测IL-1对培养的椎间盘细胞中软骨特异性基因so西和Ⅱ型胶原基因mRNA表达的调节作用。结果在IL-1浓度为0．1ng／ml、1ng／ml和10ng／ml培养24h时,其对椎间盘细胞Sox9和Ⅱ型胶原基因mRNA可起到显著的负向调控作用;10ng／ml的IL-1随着培养时间的延长对椎间盘细胞中Sox9和Ⅱ型胶原基因mRNA出现显著的负向调控作用。结论IL-1可以按照剂量及时间依赖方式负向调节椎间盘细胞Sox9和Ⅱ型胶原基因的表达。     

Surface strain on human intervertebral discs

The biomechanical functions of the internal components of the intervertebral disc are not well understood. The surface deformation of 17 human cadaveric lumbar intervertebral discs was studied by photogrammetry by adhering small optical targets to the disc surface and thereby recording the length, bulge, and vertical height of lines on the disc surface representing annular fibers. Discs were studied in pure compression, flexion and extension, axial rotation, and shear. Two definitions of a fiber were investigated: first with the end-points of the fiber on the vertebra ("bone-to-bone" definition), second, where the end points of the fiber were just before the disc vertebra junction (the "disc-only" definition). Measurements were compared with a "constant-volume" physical model and with a mathematical model of the intervertebral disc. Fiber strains were 6% or less under physiological conditions. Comparison of results from the two definitions of fiber length showed greater strains for the disc-only definition in compressive loading. Fiber strains were less than in the constant-volume model of comparable dimensions in compressive loading by a factor of about two, thus suggesting fluid loss or end-plate deformations in the physiologic conditions. The mathematical model indicated that the surface strain for intervertebral discs is very sensitive to the disc-height: diameter ratio and to fluid loss from the disc but is less sensitive to the helix angle of the fibers.     

Gene expression profile of degenerated cervical intervertebral disc tissues in rats

egenerationofintervertebraldisciscausedbymanyfactorsandisacomplicatedwebsystemofdifferentialexpressionofmanygenes .Throughstudyingthegeneexpressionprofile ,theanalysisresultsofaseriesofphysiologicalstatesofintervertebraldisccellsortissuescanbeobtainedsensitivelyandcompletely .1,2 Thesedataaremuchvaluabletotheearlydiagnosis ,thecharacteristicpreventionandtherealizationandpopularizationoftherapeuticmethodsforthisdisease ,aswellasprovideprerequisiteandimportantbasisforstudyingthetherapeuticdrugsan…     

Temporo-spatial distribution of blood vessels in human lumbar intervertebral discs

While there is consensus in the literature that blood vessels are confined to the outer anulus fibrosus of normal adult intervertebral disc, debate continues whether there is a vascular in-growths into inner parts of the intervertebral disc during degeneration. We therefore tested the hypothesis that vascular in-growth is not a distinct feature of disc degeneration. The specific endothelial cell marker CD 31 (PECAM) was used to immunohistochemically investigate 42 paraffin-embedded complete mid-sagittal human intervertebral disc sections of various ages (0–86 years) and varying extent of histomorphological degeneration. Additionally, 20 surgical disc samples from individuals (26–69 years) were included in this study. In discs of fetal to infantile age, blood vessels perforated the cartilaginous end plate and extended into the inner and outer anulus fibrosus, but not into the nucleus pulposus. In adolescents and adults, no blood vessels were seen except for the outer zone of the anulus fibrosus adjacent to the insertion to ligaments. The cartilaginous end plate remained free of vessels, except for areas with circumscribed destruction of the end plate. In advanced disc degeneration, no vessels were observed except for those few cases with complete, scar-like disc destruction. However, some rim lesions and occasionally major clefts were surrounded by a small network of capillary blood vessels extending into deeper zones of the anulus fibrosus. A subsequent morphometric analysis, revealed slightly “deeper” blood vessel extension in juvenile/adolescent discs when compared to young, mature and senile adult individuals with significantly “deeper” extension in the posterior than anterior anulus. The analysis of the surgical specimens showed that only sparse capillary blood vessels which did not extend into the nucleus pulposus even in major disc disruption. Our results show that vascular invasion deeper than the periphery was not observed during disc degeneration, which supports the hypothesis that vascular in-growth is not a distinct feature of disc degeneration. This study was supported by a grant from the AO/ASIF Foundation Switzerland (00-B72) and a grant from the AO Spine (SRN 02/103).     

基于基因表达汇编数据库的外周血中脊柱椎间盘退行性变诊断标志物的生物信息学分析

目的 通过生物信息学分析椎间盘退行性变（IDD）相关的差异表达基因（DEG）,寻找疾病的新型诊断标志物。方法 通过基因表达汇编（GEO）数据库GSE124272、GSE150408数据集下载IDD相关的外周血样本芯片数据,筛选出IDD组和正常组之间的DEG。使用DAVID在线数据库对DEG进行基因本体（GO）功能富集和京都基因与基因组百科全书（KEGG）信号通路富集,然后利用STRING在线数据库和Cytoscape软件构建蛋白质-蛋白质相互作用（PPI）网络并获取关键基因,并利用GSE23130数据集中的纤维环样本芯片数据进行验证。利用GSE124272、GSE150408数据集中的数据,采用受试者工作特征（ROC）曲线评估外周血中关键基因的诊断效能。结果 联合分析后筛选出597个DEG,包含363个上调基因和234个下调基因。GO功能富集分析发现DEG主要参与细胞黏附、细胞凋亡、趋化作用和细胞迁移等功能,KEGG分析发现DEG主要参与细胞外基质受体相互作用和癌症中的信号通路。PPI网络分析筛选出17个关键基因,经验证获得RBMX、EEF1A1、SSR1和POLR2C这4个基因,ROC曲线分析显示这4个基因对IDD诊断效能显著,曲线下面积分别为0.763、0.741、0.710、0.702。结论 RBMX、EEF1A1、SSR1和POLR2C或可成为IDD的新型诊断标志物,为该病进一步的功能研究提供理论依据。     

The compressive creep properties of normal and degenerated murine intervertebral discs.

Identifying mechanisms by which degeneration alters intervertebral disc material properties and biomechanical behavior is important for clarifying back pain risk factors as well as for evaluating the efficacy of novel interventions. Our goal was to quantify and characterize degeneration-dependent changes in the disc's response to compression using a previously established murine model of disc degeneration. We performed compressive creep tests on normal and degenerated murine intervertebral discs and parameterized the biomechanical response using a previously established fluid-transport model. Using a series of biochemical and histological assays, we sought to determine how biomechanical alterations were attributable to degeneration-related changes in tissue morphology. We observed that with moderate degeneration, discs lost height (mean+/-std. dev. of 0.44+/-0.01 vs. 0.36+/-0.01 mm, p<0.0001), increased in proteoglycan content (31+/-4 vs. 43+/-2 microg/ml of extract, p<0.0002), became less stiff (2.17+/-0.66 vs. 1.56+/-0.44 MPa, p<0.053), and crept more. Model results suggested that the increased creep response was mainly due to a diminished strain-dependent nuclear swelling pressure. We also noted that the model-derived tissue properties varied with the applied load magnitude for both normal and degenerated discs. Overall, our data demonstrate that architectural remodeling stimulated by excessive loading diminishes the disc's ability to resist compression. These results are similar to degeneration-dependent changes reported for human discs.     

骨形态发生蛋白-2与椎间盘细胞Sox9 和Ⅱ型胶原基因的调控关系

目的：探讨骨形态发生蛋白-2（BMP-2）对椎问盘细胞Sox9和Ⅱ型胶原基因表达的调节作用。方法：应用逆转录聚合酶链反应和Western印迹检测不同浓度BMP-2对培养的人椎间盘细胞中软骨特异性基因Sox9和Ⅱ型胶原基因表达的调节作用。结果：BMP-2浓度为100ng／ml和1000ng／ml时，椎间盘细胞中Sox9和Ⅱ型胶原基因mRNA表达与0n∥ml组（对照组）相比明显增加（P〈0．05）；在蛋白水平的检测中，也得到了一致的结果。结论：BMP-2可以按照剂量依赖方式正向调节椎间盘细胞Sox9和Ⅱ型胶原基因的表达，提示BMP-2可能对退变早期的椎间盘具有修复功能。     

骨形态发生蛋白-2对椎间盘细胞软骨特异性基因 mRNA的调节作用

目的探讨骨形态发生蛋白(BMP)-2对椎间盘细胞软骨特异性基因Sox9、Ⅱ型胶原和蛋白聚糖基因的调控作用.方法应用逆转录-聚合酶链反应(RT-PCR)技术检测BMP-2对培养的人椎间盘细胞中Sox9、Ⅱ型胶原和蛋白聚糖基因 mRNA表达的调控作用.结果在BMP-2浓度为100 μg/L(0.149±0.006,P<0.05)和1 000 μg/L(0.163±0.006,P<0.01)时,其对椎间盘细胞中Sox9基因 mRNA可起到显著的正向调控作用;在此浓度下,它也可以对Ⅱ型胶原和蛋白聚糖基因mRNA起到正向调控作用.结论 BMP-2可以按照剂量依赖方式正向调控椎间盘细胞中Sox9、Ⅱ型胶原和蛋白聚糖基因的表达.