凋亡及免疫激活相关基因在先兆子痫胎盘中的表达

目的：探讨细胞凋亡相关基因在先兆子痫胎盘组织中的表达谱及其影响机制。方法：采用分别包含220余种人类细胞因子相关基因或人类环境激素相关基因cDNA片段的基因芯片，检测严格配伍的先兆子痫和正常胎盘组织中基因表达谱的差异。结果：先兆子痫胎盘中与细胞增殖周期调控及凋亡相关的多种基因表达发生了变化，且大多数呈现表达增强。此外，与免疫系统激活有关的多条基因在先兆子痫胎盘中的表达也比正常胎盘高。结论：胎盘组织中多种与细胞凋亡调节有关的基因表达异常与先兆子痫的病理发生有关，而免疫系统激活也可能是其原因之一。     

代谢相关基因在先兆子痫胎盘中的表达情况分析

目的探讨细胞代谢相关基因在先兆子痫胎盘组织中的表达变化及其影响机制。方法使用含12000个与代谢、凋亡、细胞粘附、信号传导、转录因子等有关基因的cDNA表达谱芯片,检测4例重度妊娠高血压疾病子痫前期及正常的胎盘组织的基因表达谱差异,并差异表达的细胞代谢相关基因进行了Northern验证。结果4先兆子痫胎盘中同时有44种基因表达发生了变化,其中糖原磷酸化酶（GP—M）基因、瘦素（1eptin）基因、脂蛋白酯酶（LPL）基因及糖代谢相关基因（Glucose transporter）表达增强,核苷酸代谢基因（CD73）与能量代谢调节基因（Creatine kinase B）表达下降。结论多种基因表达异常与先兆子痫的病理发生有关,细胞代谢相关基因表达异常可能是血管内皮损伤的原因之一。     

透骨消痛胶囊含药血清对膝骨关节炎关节滑膜细胞uPA系统及炎性因子的影响

背景：透骨消痛胶囊是治疗骨性关节炎的临床验方,作用机制尚未完全阐明。尿激酶型纤溶酶原激活系统参与软骨细胞外基质降解和滑膜增生,在骨性关节炎发生、发展中起重要作用。目的：观察透骨消痛胶囊含药血清对体外培养的膝骨性关节炎大鼠滑膜细胞表达尿激酶型纤溶酶原激活剂、尿激酶型纤溶酶原激活剂受体、纤溶酶原激活剂抑制剂、基质金属蛋白酶3、白细胞介素1β和肿瘤坏死因子α的影响,探讨透骨消痛胶囊防治膝骨性关节炎的作用机制。方法：采用4%木瓜蛋白酶注射关节腔复制大鼠膝骨性关节炎模型;胶原酶消化法原代培养正常滑膜细胞与骨性关节炎滑膜细胞,将所培养滑膜细胞分为正常组、模型组和透骨消痛胶囊组,前2组细胞用空白血清培养,透骨消炎胶囊用含药血清培养;透骨消痛胶囊含药血清作用骨性关节炎滑膜细胞72 h后,采用Western Blot法检测滑膜细胞中尿激酶型纤溶酶原激活剂、尿激酶型纤溶酶原激活剂受体、纤溶酶原激活剂抑制剂、基质金属蛋白酶3、白细胞介素1β和肿瘤坏死因子α蛋白的表达。结果与结论：透骨消痛胶囊含药血清组尿激酶型纤溶酶原激活剂、尿激酶型纤溶酶原激活剂受体、基质金属蛋白酶3、白细胞介素1β和肿瘤坏死因子α蛋白表达量均降低,纤溶酶原激活剂抑制剂表达量升高,与模型组相比均有显著性差异,提示透骨消痛胶囊治疗骨性关节炎的部分作用机制是有效抑制骨性关节炎滑膜细胞尿激酶型纤溶酶原激活剂、尿激酶型纤溶酶原激活剂受体、基质金属蛋白酶3、白细胞介素1β和肿瘤坏死因子α的表达,提高纤溶酶原激活剂抑制剂的表达。中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

先兆子痫胎盘的基因表达谱研究

目的： 利用cDNA芯片分析先兆子痫胎盘中基因表达的变化情况,寻找新的先兆子痫相关基因。方法: 建立含12 000个与代谢、凋亡、细胞黏附、信号转导、转录因子等有关基因的cDNA表达谱芯片,将先兆子痫及正常胎盘mRNA与cDNA芯片进行杂交,得到先兆子痫的基因表达谱;对部分差异表达基因进行Northern验证。结果: 在4例先兆子痫胎盘组织中发现均有差异表达的基因44个,其中表达上调有30个,表达下调有14个,Northern杂交鉴定的结果与芯片结果相符合。结论: 重度妊娠高血压疾病的基因表达谱存在明显差异,差异的基因可能涉及信号转导、细胞代谢、炎性细胞因子等方面,这些基因可能与妊娠高血压病的发病相关。     

MMP9及TIMP1 mRNA在先兆子痫患者胎盘中的表达及其意义

目的检测基质金属蛋白酶9及其抑制物1的mRNA(MMP9mRNA、TIMP1mRNA)在先兆子痫患者胎盘中的表达,以探求MMP9在先兆子痫发病机制中的意义.方法用半定量逆转录聚合酶链反应(RT-PCR)技术检测正常妊娠(10例),先兆子痫(22例)剖宫产后胎盘中MMP9、TIMP1基因在转录水平的表达.结果重度先兆子痫胎盘中MMP9mRNA表达量降低(0.05±0.04),TIMP1mRNA表达量增加,MMP9/TIMP1比值下降(0.04±0.03),与轻度先兆子痫及正常对照组比较有明显意义.结论 MMP9表达在基因转录水平就已经下调,同时因TIMP1表达增强其活性进一步被抑制,因而在先兆子痫的发生和发展中有重要意义.     

应用基因芯片研究代谢相关基因在先兆子痫胎盘中的表达

目的:研究部分代谢相关基因在先兆子痫胎盘组织中的表达变化。方法:采用包含220余种人类环境激素相关基因cDNA片段的基因芯片,对经过严格配伍的正常和先兆子痫胎盘组织中基因表达谱的差异进行检测。结果:先兆子痫胎盘中与氧化还原代谢相关的基因(GenBank:U78168、X16699、D32143、AL035079、AF069668、X53463、J03746、X02317、X91247等)表达增高。另外,与激素等其它代谢相关的一些基因(如GenBank:NM_-001718、Z22535、M38180、M76665、X75252、J03258、U56725)在先兆子痫胎盘中的表达也增强。结论:胎盘内基因表达的变化与先兆子痫患者体内多种代谢改变有关。     

白细胞介素及肿瘤坏死因子基因超家族在先兆子痫胎盘中的表达

为探讨白细胞介素和肿瘤坏死因子 (受体 )超家族基因表达与先兆子痫病理发生的关系 ,以包含 2 4 3种人类细胞因子相关基因cDNA片段的基因芯片 ,检测严格配对的先兆子痫和正常胎盘组织中基因表达谱的差异。结果显示受检的白细胞介素和 (或 )白细胞介素受体基因共 2 2种 ,绝大多数基因在先兆子痫胎盘中的表达增强 ,而IL 2受体 (IL 2Rα )基因 (Gen Bank :X0 10 5 7)在先兆子痫胎盘中的表达低于正常胎盘。肿瘤坏死因子 (GenBank :X0 2 910 )及其配体 (GenBank :U0 3398、U375 18、AF0 5 3712、AF0 5 5 872 )、受体 (GenBank :X6 0 5 92、X6 3717、M835 5 4、AF0 16 2 6 6、AF0 16 2 6 7、U812 32 )等 10余种肿瘤坏死因子 (受体 )超家族基因在先兆子痫胎盘中的表达也较高。说明 ,白细胞介素及肿瘤坏死因子 (受体 )基因超家族的高表达可能与先兆子痫的病理发生关系密切     

脑源性神经营养因子对细胞外蛋白水解酶激活作用的研究

 目的：研究脑源性神经营养因子 (BDNF) 对细胞外蛋白水解酶表达和激活作用的影响。 方法：体外分离并培养人脐静脉内皮细胞（HUVEC），RT-PCR法检测HUVEC基质金属蛋白酶MMP-2 、MMP-9 和基质金属蛋白酶组织抑制剂TIMP-1、TIMP-2 mRNA的表达，明胶酶谱检测MMP-2和MMP-9蛋白酶活性，纤维蛋白酶谱检测尿激酶型纤溶酶原激活剂（uPA）蛋白酶活性，Western blotting检测uPA、纤溶酶原激活剂抑制剂（PAI）、TIMP-1及TIMP-2表达。 结果：在对HUVEC增殖无明显促进作用的浓度范围内，BDNF可促进无血清培养的HUVEC MMP-2和MMP-9 mRNA表达,并可促进MMP-2和MMP-9酶原的激活产生活性明胶酶，BDNF对TIMP-1和TIMP-2的表达无明显影响。BDNF以浓度和时间依赖性方式上调HUVEC uPA和PAI-1的表达，并可促进uPA的活性。 结论：BDNF可激活MMPs和uPA/PAI相关的蛋白级联。     

RECK、基质金属蛋白酶-9和血管内皮生长因子在子痫前期患者胎盘组织中的表达

目的检测子痫前期患者胎盘组织中RECK、基质金属蛋白酶-9(MMP-9)、血管内皮生长因子(VEGF)的基因表达,探讨它们与胎盘滋养细胞浸润过程的调控关系。方法采用RT-PCR、Western blotting、免疫组织化学检测120例妊娠足月剖宫产(正常妊娠、轻度子痫前期、中度子痫前期、重度子痫前期各30例)胎盘组织中RECK、MMP-9、VEGF的基因表达。结果 3组子痫前期患者胎盘组织中MMP-9及VEGF的mRNA表达水平均显著低于正常妊娠组(P0.05);重度子痫前期组中RECK mRNA的表达显著高于正常妊娠组(P0.05);3组子痫前期患者胎盘组织中MMP-9及VEGF蛋白表达均显著低于正常妊娠组(P0.05),中度和重度子痫前期组中RECK蛋白表达显著高于正常妊娠组(P0.05)。结论子痫前期患者胎盘中RECK与MMP-9、VEGF之间存在负相关性,它们可能参与了胎盘滋养细胞浅浸润过程的调控。     

基质金属蛋白酶-9和血管内皮生长因子在子痫前期患者胎盘组织中的表达

目的 检测子痫前期患者胎盘组织中RECK、基质金属蛋白酶-9（MMP-9）、血管内皮生长因子（VEGF）的基因表达,探讨它们与胎盘滋养细胞浸润过程的调控关系。 方法 采用RT-PCR、Western blotting、免疫组织化学检测120例妊娠足月剖宫产（正常妊娠、轻度子痫前期、中度子痫前期、重度子痫前期各30例）胎盘组织中RECK、MMP-9、VEGF的基因表达。结果 3组子痫前期患者胎盘组织中MMP-9及VEGF的mRNA表达水平均显著低于正常妊娠组（P<0.05）;重度子痫前期组中RECK mRNA的表达显著高于正常妊娠组（P<0.05）;3组子痫前期患者胎盘组织中MMP-9及VEGF蛋白表达均显著低于正常妊娠组（P<0.05）,中度和重度子痫前期组中RECK蛋白表达显著高于正常妊娠组（P<0.05）。结论 子痫前期患者胎盘中RECK与MMP-9、VEGF之间存在负相关性,它们可能参与了胎盘滋养细胞浅浸润过程的调控。     

Expression profile of trophoblast invasion-associated genes in the pre-eclamptic placenta

The primary pathology of pre-eclampsia is thought be a defect in placentation due to failure of trophoblast invasion. Here, we aim to identify the expression profile of invasion-associated genes in the pre-eclamptic placenta. Messenger RNA (mRNA) expression levels of extracellular matrix molecule-related genes in five pre-eclamptic placentas and in five strictly matched normal placentas were assayed using complementary DNA (cDNA) microarrays representing over 220 human cytokine-associated or hormone-associated genes. Results demonstrated greater than two-fold higher expression of 18 extracellular matrix molecule genes, including cadherin, collagen, integrin and selectin, in the pre-eclamptic placenta. Extracellular matrix molecule degradation-related genes, including matrix metalloproteinase (MMP)-10, MMP-13, MMP-15, tissue inhibitor of metalloproteinase (TIMP)-2, TIMP-3, plasminogen and plasminogen activator, were also highly expressed in the pre-eclamptic placenta, compared to the normal placenta. Results suggest that the abnormal expression profiles of extracellular matrix molecules and degrading proteinases might be associated with the pathogenesis of pre-eclampsia.     

早孕和足月滋养细胞中侵袭相关性基因的表达变化

目的：探讨早孕和足月滋养细胞侵袭能力差异的机制。方法：分离并在体外培养早孕和足月的滋养细胞为，采用RT-PCR法，检测其侵袭相关性基因的表达。结果：早孕滋养细胞表达MMP-2、MMP-9、TIMP-1和UPA；而晚孕滋养细胞表达MMP-2、TIMP-1、TIMP-2和PAI。晚孕滋养细胞TTMP—1的表达显著高于早孕滋养细胞，而MMP-2的表达则无明显变化。结论：MMP-9等蛋白酶及其抑制因子表达的变化，可能是影响滋养细胞侵袭性的重要原因。     

MT1‐MMP evaluation in neointimal hyperplasia in the late follow‐up after prosthesis implantation

Vascular surgical interventions are often burdened with late complications, including thrombosis or restenosis. The latter is generally caused by neointimal hyperplasia. Although extracellular matrix (ECM) remodelling is an important part of neointima formation, this process is not clearly understood. The aim of the study was to assess the content and activity of membrane‐type 1 matrix metalloproteinase in human neointima in the late stages of its development. Matrix metalloproteinase‐2 and tissue inhibitor of matrix metalloproteinase‐2 were also evaluated. The research was performed on neointima samples collected during secondary vascular interventions from patients with chronic limb ischaemia who developed vascular occlusion at 6‐18 months after aorto/ilio‐femoral bypass grafting. The control material consisted of segments of femoral arteries collected from organ donors. Western blot and/or ELISA were used for the determination of MT1‐MMP and TIMP‐2 expression. The activity of MT1‐MMP was measured by fluorometric assay and that of MMP‐2 by zymography. We demonstrated significantly increased MT1‐MMP protein content in neointima when compared to normal arteries. However, the activity of MT1‐MMP was significantly lower in neointima than in control samples. The decreased MT1‐MMP activity was concomitant with reduced activity of MMP‐2. The TIMP‐2 protein levels in neointima and normal arteries were not significantly different. The results of our study suggest that the reduced activity of MT1‐MMP and consequently MMP‐2 in human neointima may play a role in decreased degradation of ECM components and thus promote neointimal overgrowth.     

Could the Defects in the Endometrial Extracellular Matrix During the Implantation Be a Cause for Impaired fertility?

PROBLEM: Assessment of concentration of the metalloproteinase 9 (MMP9), tissue inhibitor of metalloproteinases 1 (TIMP1), urokinase plasminogen activator (uPA) and its receptor (uPAR) and leukaemia inhibitory factor (LIF) in the uterine fluid in women with idiopathic infertility and unknown cause for recurrent miscarriage. METHOD OF STUDY: Uterine fluid obtained from patients with idiopathic infertility (n = 13), patients with unknown cause of recurrent miscarriages (n = 16) and healthy fertile women (n = 14). The assessment of MMP9, TIMP1, uPA, uPAR and LIF were performed with enzyme-linked immunosorbent assay. RESULTS: Statistically lower expression of MMP9 and TIMP1 in uterine fluid during implantation window in women with infertility and recurrent miscarriage of unknown cause. CONCLUSION: Changed turnover of the extracellular matrix might be a cause of improper endometrial transition and in effect cause faulty implantation.     

Vascular endothelial growth factor expression in the basal subtype of breast carcinoma

The recognition of subtypes of breast carcinomas based on their molecular features has brought new perspectives in breast cancer research. Some key regulators of angiogenesis and tumor infiltration were evaluated in breast carcinomas of basal phenotype (cytokeratin [CK]5+). Immunohistochemical analysis with 14 primary antibodies was performed in 100 formalin-fixed, paraffin-embedded samples of invasive ductal carcinomas. CK5 correlated with indicators of poor outcome, including precocious age, high histologic grade, lymph node positivity, advanced pathologic stage, negativity for hormonal receptors, and a high proliferative rate (Ki-67 labeling index). CK5 also correlated with vascular endothelial growth factor (VEGF) expression but not with the microvessel density. Considering that VEGF-overexpressing neoplastic mammary cells display increased proliferative activity in vitro regardless of the angiogenic effect of VEGF, the differential expression of VEGF might contribute to the more aggressive behavior of these neoplasms. CK5 correlated with tissue inhibitor of metalloproteinase (TIMP)-1, but not matrix metalloproteinase (MMP)-1, MMP-2, extracellular matrix metalloproteinase inducer, TIMP-2 or plasminogen activator inhibitor, indicating that antiproteolytic stimuli might be preponderant in these neoplasms.     

ACE inhibition reduces activity of the plasminogen/plasmin and MMP systems in the brain of spontaneous hypertensive stroke-prone rats

The spontaneously hypertensive stroke-prone rat (SHR-SP) is an experimental model of malignant hypertension which lead to secondary alterations of the extracellular matrix. Our aim was to determine ACE-inhibitor related changes of proteases involved in the reconstruction of the extracellular matrix in the brain. Twelve SHR-SP rats were randomized into two groups. Each group was treated with either an antihypertensive dose of ramipril or placebo for 6 months. Brain tissue plasminogen activator (t-PA) and urokinase (u-PA) were quantified by using casein-dependent plasminogen zymography, matrix metalloproteinase (MMP)-2 and MMP-9, by MMP-zymography, and tissue inhibitor of MMP (TIMP)-1 and -2, by reverse zymography. The amounts of u-PA, t-PA, and MMPs were significantly reduced in animals treated with ACE inhibitor. Plasminogen zymography showed a 39% reduction of u-PA in the basal ganglia (p < 0.0001); t-PA expression was reduced by 26% in the cortex and by 33% in the basal ganglia (p < 0.0001). MMP-2 expression was reduced by 15% in the cortex (p < 0.05) and by 10% in the basal ganglia (p < 0.05); MMP-9 expression significantly decreased by 37% in the cortex and by 25% in the basal ganglia (p < 0.0001 each). No differences were observed in the amount of TIMP-1 or TIMP-2. These findings provide new insights into the biochemical mechanisms underlying extracellular matrix proliferation and its modulation by ACE inhibitors. Therapeutic alterations that influence the proteolytic systems might prove important in the prevention of extracellular matrix accumulation and secondary microvascular vessel wall changes.     

黄芪多糖对颈椎病模型大鼠颈椎间盘纤维环MMP2和MMP9表达的影响

目的:探究黄芪多糖(AP)对颈椎病模型大鼠颈椎间盘纤维环中基质金属蛋白酶2(MMP2)和MMP9表达的影响。方法:建立动静力失衡性颈椎间盘退变大鼠模型,将造模成功大鼠随机分为模型组(M组)及AP低、高剂量处理组(L-AP组和H-AP组),以假手术组大鼠作为阴性对照组(NC组),另取各组大鼠颈椎间盘纤维环组织细胞进行原代细胞培养。应用HE染色和藏红O染色进行组织学分析。应用免疫组织化学染色、Western blot和RT-qPCR法检测MMP2、MMP9、金属蛋白酶组织抑制物2(TIMP2)和Ⅳ型胶原(collagenⅣ)的mRNA和蛋白表达。应用细胞-胶原黏附实验检测纤维环细胞-胶原黏附作用。结果:M组大鼠椎间盘出现退行性病变,黄芪多糖能够改善颈椎病大鼠椎间盘退行性病变。与NC组相比,M组大鼠纤维环组织中MMP2和MMP9表达水平显著增加,而TIMP2和collagenⅣ表达水平均显著降低(P 0. 05);与M组相比,L-AP组和H-AP组的MMP2和MMP9表达水平显著降低,而TIMP2和collagenⅣ的表达水平均显著增加(P 0. 05)。M组大鼠纤维环细胞-胶原黏附作用显著低于NC组(P 0. 05);与M组相比,L-AP组和H-AP组的纤维环细胞-胶原黏附作用均显著上升(P 0. 05)。与NC组相比,M组纤维环细胞中的MMP2和MMP9表达水平显著增加,而TIMP2和collagenⅣ的表达水平均显著降低(P 0. 05);与M组相比,L-AP组和H-AP组纤维环细胞中的MMP2和MMP9表达水平显著降低,而TIMP2和collagenⅣ表达水平均显著增加(P 0. 05)。结论:黄芪多糖能够抑制颈椎病模型大鼠纤维环组织中MMP2和MMP9表达,调节细胞外基质中MMPs与TIMPs的动态平衡,从而抑制MMPs对椎间盘基质中胶原的降解,在椎间盘退变的治疗中具有潜在研究价值。     

牙周炎牙龈组织MMP-2、TIMP-1的免疫组化研究

目的　探讨来源于宿主的基质金属蛋白酶MMPs及其抑制因子在牙周炎发病中的作用机制。方法　本研究利用免疫组化方法检测 15例牙周炎患者牙龈组织中和 4例健康牙龈中基质金属蛋白酶 2(MMP 2 )、组织抑制因子 1(TIMP 1)的蛋白表达。结果　 15例成人牙周炎组牙龈组织中MMP 2在上皮内及上皮下的炎性组织中均有阳性表达 ,而且阳性率明显高于正常牙龈 (P <0 .0 5 ) ;TIMP 1在牙周炎以及正常的牙龈组织中表达无明显差异。结论　本研究提示基质金属蛋白酶MMP 2参与牙周组织破坏过程 ,参与牙周炎的病理生理机制