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1.
骨形态发生蛋白-7(BMP-7)是具有强诱骨活性的蛋白质因子,已通过基因工程技术在体外得到表达,较长时间以来不断被应用于骨损伤疾病的研究,得到了确切的治疗效果。通过载体将BMP-7基因转入真核细胞,与生物聚合载体复合后植入体内,能表达并分泌活性的BMP-7,诱导骨细胞的生成,促进骨组织的修复,成为一种新的有效的治疗手段。  相似文献   

2.
多种基因工程抗体、酶、激素、血浆蛋白和疫苗等都已在植物的叶、茎、根、果实、种子以及植物细胞和器官中得到表达 ,然而提取和纯化始终是大规模利用植物生产重组蛋白的主要障碍 .Borisjuk和Komamytsky等 (1999年和 2 0 0 0年 )依据内质网和内质网信号肽在蛋白质合成中的作用 ,把 3种重组蛋白 ,嗜热细菌来源的木聚糖酶、水母的绿色荧光蛋白和人胎盘分泌的碱性磷酸酶 (SEAP) ,定位到质外体中 ,通过植物根分泌和叶分泌途径获得表达 ,从而建立了 2种新的重组蛋白表达系统———植物根分泌和叶分泌 ,简化了分离和纯化程…  相似文献   

3.
可应用于骨组织修复的具有生物活性的复合医用材料——人工骨,采用60%FDA批准的高分子材料高分子材料聚乙内酯多元醇和40%陶瓷原料磷酸三钙用融合塑型技术,生产出物理性状类似骨组织的人工骨,再以交联的方式将骨形态发生蛋白BMP-2,修饰复合材料表面,增强复合材料的生物成骨活性,可促进骨组织再生。  相似文献   

4.
骨形态形成蛋白因子Ⅱ(BMP-2)属转化生长因子TGF-β超家族成员,具有活性最高和能单独诱导骨形成的特性。BMP-2能促进成骨细胞分化和增加成骨细胞标志基因的表达,这在成骨细胞分化过程中起非常关键的作用,最终促进骨形成。因此BMP-2可作为治疗骨质疏松症药物的新作用靶点。目前,已证明可上调BMP-2表达的上调剂多为Statin类药物。它们多通过抑制Rho和Rho激酶的活性;增加游离的有活性的eNOS;抑制HMG-CoA还原酶的活性等方面促骨形成。研究Statin类药物促BMP-2表达上调的机理,可为进一步寻找更有效的治疗骨质疏松症药物提供新靶点和新思路。  相似文献   

5.
目的:研究骨碎补总黄酮对切除卵巢的雌性大鼠血清骨钙素和骨形态蛋白-2(bone morphogenetic protein-2,BMP-2)在骨组织中表达的影响。方法:取3月龄雌性大鼠30只,随机分成假手术组、给药组和空白对照组。假手术组仅去除卵巢周围脂肪,给药组在去除卵巢后连续15周灌胃给服强骨胶囊,其余给等量自来水。15周后处死所有大鼠,取股骨中段组织,行BMP-2免疫组化染色,镜下观察染色标本摄像并用Image图形分析软件分析各标本阳性表达率。另取大鼠血清以放免法测定其血清骨钙素含量。结果:免疫组化染色后,图片分析显示给药组和假手术组bmp-2阳性率明显高于对照组(p<0.05);给药组和假手术组血清骨钙素的表达明显高于对照组(p<0.05)。结论:骨碎补总黄酮对卵巢切除大鼠的BMP-2和血清骨钙素表达有促进作用。  相似文献   

6.
骨形成蛋白(Bone Morphogenetic Protein,BMP)是一类能诱导异位骨及软骨形成,并在动物的发育和分化中起作用的蛋白质[1,2,3]。自Urist及其同事发现骨形成蛋白以来4。已对8种人的BMP进行了克隆,除BMP-1外[5],BMP-2至BMP-8均与TGF-β家族相关,它们能诱导细胞分化,促进骨、软骨及牙本质的形成[1,6]。并在发育、分化和形成过程中起重要作用。最新的研究认为BMP-1是一种胶原蛋白酶[7],进一步揭示了BMP家族成员的生物学作用。人的BMP-3基因定位于第4染色体上,BMP-3蛋白由472个氨基酸组成,包括N端的信号肽、中间的前肽及C端的成熟肽三部分。BMP-3的C末端与MBP-2A及BMP-2B有49%的序列相同[5]。本实验室曾检测了BMP-3和BMP-5在不同 组织和细胞中的表达情况,发现它们在一些与骨形成无关的组织和细胞中均有表达,说明了BMP在动物和人中有着其他重要的作用[8]。在此基础上,我们对BMP-3进行了克隆及在大肠杆菌中高效表达BMP-3-GST融合蛋白,并用Western印迹证明了其活性。  相似文献   

7.
目的鉴定在大肠杆菌中表达的重组人骨形态发生蛋白2(recombinant human bone morphogeneticprotein-2,rhBMP-2)的生物学活性。方法构建重组人BMP-2的原核表达质粒,在大肠杆菌中诱导表达目的蛋白,纯化复性后,采用细胞及动物实验对其生物活性进行鉴定。结果细胞实验表明,重组人BMP-2具有诱导MC3T3-E1前成骨细胞向成骨细胞分化的特性。动物实验显示,重组人BMP-2在大鼠肌肉内具有诱导异位成骨功能。结论制备的重组人BMP-2具有较好的生物学活性。  相似文献   

8.
目的:探讨骨形态发生蛋白( BMP-7)在前列腺癌组织中的表达及其与临床分期之间的关系.方法:应用免疫印迹法检测30例前列腺癌患者及30例前列腺良性增生患者前列腺组织中BMP-7的表达情况.结果:前列腺癌组织中BMP-7的表达显著高于前列腺良性增生组织,且BMP-7的表达随前列腺癌的临床分期、Gleason分级增高而增加.结论:BMP-7在前列腺癌中的表达明显增高,其表达量与临床分期相关,前列腺癌组织中BMP-7的表达增高提示预后不佳.  相似文献   

9.
植物谷胱甘肽过氧化物酶(glutathione peroxidase,GPX)是清除体内活性氧的一种关键酶,在植物抗逆反应中发挥重要作用.本研究从水稻中克隆到2个GPX基因,分别为OsGPX3和OsGPX4.OsGPX3和OsGPX4分别编码238和234个氨基酸组成的蛋白质,预测分子量分别是25.84 kD和25.07 kD.两个基因都包含5个内含子,但是两个基因所对应的内含子长度具有较大变异.组织表达谱分析发现这2个基因在根、茎、叶和叶鞘中均表达,是组成型表达基因.在大肠杆菌中表达并纯化了这2个基因的重组蛋白,酶活性分析显示OsGPX3和OsGPX4蛋白对底物H2O2、tBOOH和COOH具有较高活性,但是OsGPX3对3种底物的活性均高于OsGPX4,蛋白质酶活性的差异预示着这2个基因可能存在功能上的分化.  相似文献   

10.
目的:探讨血府逐瘀汤对股骨颈骨折术后患者转化生长因子β(TGF-β)、血管内皮生长因子(VEGF)、骨形成蛋白-2(BMP-2)表达的影响。方法:选择2014年2月至2014年8月于我院接受治疗的股骨颈骨折患者80例,随机分为实验组和对照组。对照组行常规治疗方案,实验组在常规治疗基础上加用血府逐瘀汤。观察并比较两组患者治疗前后自觉疼痛程度及骨折处骨组织TGF-β、VEGF及BMP-2表达的变化。结果:治疗后,两组患者harris评分均高于治疗前,实验组显著高于对照组(P0.05)。两组患者的骨折处骨组织TGF-β、VEGF、BMP-2水平均高于治疗前,实验组高于对照组,差异均有统计学意义(P0.05)。结论:血府逐瘀汤能有效提高股骨颈骨折患者骨折处骨组织TGF-β、VEGF、BMP-2的表达,促进股骨颈骨折的恢复,值得临床应用和推广。  相似文献   

11.
The bone morphogenetic proteins (BMPs) are a family of growth factors that regulate the development of bone. BMP-2 is the most effective in the induction of bone tissue. A large amount of BMP-2 is needed for both bone tissue engineering research and clinical application. Thus, an effective way is necessary to produce sufficient BMP-2 protein. With the advance in plant biotechnology, transgenic plants have been targeted as a bioreactor to produce desired recombinant proteins. Here, the expression of recombinant human bmp-2 gene (rhbmp-2) was studied in tobacco plants using gus as a reporter gene. The difference of expression levels in root, stem and leaf tissues was analyzed by GUS activity assay, semi-quantitive RT-PCR and western blotting. The results indicated that the expression levels of fusion protein in root and stem tissues were significantly higher than those in leaf tissue. For the protein compositions in root and stem tissues were simpler than those in leaf tissue, this suggested that the purification process with root and stem tissues would potentially be easier.  相似文献   

12.
Proteomic analysis of rice leaf, stem and root tissues during growth course   总被引:4,自引:0,他引:4  
Nozu Y  Tsugita A  Kamijo K 《Proteomics》2006,6(12):3665-3670
Rice proteins were isolated from leaf, stem and root tissues, harvesting at 1, 2, 4, 8 and 10 weeks after budding. Each tissue of each age was separately pulverized in liquid nitrogen, and the resulted tissue powders were suspended in 10% TCA-acetone and followed by acetone suspension to precipitate at low temperature, which resulted in the tissue-specific and age-specific protein mixture. The protein mixtures were separated by 2-DE using polyacrylamide gels (26 x 20 cm). The protein spots were identified by N-terminal sequence analysis and by MALDI and LC-MS/MS analyses after in-gel tryptic digestion. From a total of 4532 spots, 676 unique proteins were identified, of which 80 proteins (12%) were observed in all three tissues: leaf, stem and root. In addition, 45 (7%) were common in leaf and stem, 57 (8%) in stem and root, and 10 (2%) proteins in root and leaf. Also 141 unique proteins (21%) were observed only for leaf, 96 (14%) for stem, and 247 (36%) for root tissue. Proteins playing a role for photosynthesis and energy production were most abundant in leaf and stem, and those for cell defense were rich in roots.  相似文献   

13.
Expression of the Cry2Aa2 protein was targeted specifically to the green tissues of transgenic tobacco Nicotiana tabacum cv. Xanthi plants. This deployment was achieved by using the promoter region of the gene encoding the Solanum tuberosum leaf and stem specific (ST-LS1) protein. The accumulated levels of toxin in the leaves were found to be effective in achieving 100 mortality of Heliothis virescens larvae. The levels of Cry2Aa2 expression in the leaves of these transgenic plants were up to 0.21 of the total soluble proteins. Bioassays with R1 transgenic plants indicated the inheritance of cry2Aa2 in the progeny plants. Tissue-specific expression of the Bt toxin in transgenic plants may help in controlling the potential occurrence of insect resistance by limiting the amount of toxin to only predated tissues. The results reported here validate the use of the ST-LS1 gene promoter for a targeted expression of Bt toxins in green tissues of plants.  相似文献   

14.
Bone morphogenetic proteins (BMPs) are cytokines from the TGF-β superfamily, with important roles during embryonic development and in the induction of bone and cartilage tissue differentiation in the adult body. In this contribution, we report the expression of recombinant human BMP-4, BMP-9, BMP-10, BMP-11 (or growth differentiation factor-11, GDF-11) and BMP-14 (GDF-5), using Escherichia coli pET-25b vector. BMPs were overexpressed, purified by affinity his-tag chromatography and shown to induce the expression of early markers of bone differentiation (e.g. smad-1, smad-5, runx2/cbfa1, dlx5, osterix, osteopontin, bone sialoprotein and alkaline phosphatase) in C2C12 cells and in human adipose stem cells. The described approach is a promising method for producing large amounts of different recombinant BMPs that show potential for novel biomedical applications.  相似文献   

15.
Osteogenesis versus chondrogenesis by BMP-2 and BMP-7 in adipose stem cells   总被引:7,自引:0,他引:7  
Bone morphogenetic proteins (BMPs) initiate, promote, and maintain chondrogenesis and osteogenesis. We hypothesize that BMP-2 induces an osteogenic, and BMP-7 a chondrogenic phenotype in adipose tissue-derived mesenchymal stem cells (AT-MSCs). We compared the effects of a short 15min BMP-2 or BMP-7 (10ng/ml) treatment on osteogenic and chondrogenic differentiation of AT-MSCs. Gene expression was studied 4 and 14 days after BMP-treatment. At day 4 BMP-2, but not BMP-7, stimulated runx-2 and osteopontin gene expression, and at day 14 BMP-7 down-regulated expression of these genes. At day 4 BMP-2 and BMP-7 stimulated biglycan gene expression, which was down-regulated by BMP-7 at day 14. BMP-7 stimulated aggrecan gene expression at day 14. Our data indicate that BMP-2 treatment for 15min induces osteogenic differentiation, whereas BMP-7 stimulates a chondrogenic phenotype of AT-MSCs. Therefore, AT-MSCs triggered for only 15min with BMP-2 or BMP-7 provide a feasible tool for bone and cartilage tissue engineering.  相似文献   

16.
Kwak MS  Oh MJ  Lee SW  Shin JS  Paek KH  Bae JM 《Plant cell reports》2007,26(8):1253-1262
To develop a strong constitutive gene expression system, the activities of ibAGP1 promoter and its transit peptide were investigated using transgenic Arabidopsis and a GUS reporter gene. The ibAGP1 promoter directed GUS expression in almost entire tissues including rosette leaf, inflorescence stem, inflorescence, cauline leaf and root, suggesting that the ibAGP1 promoter is a constitutive promoter. GUS expression mediated by ibAGP1 promoter was weaker than that by CaMV35S promoter in all tissue types, but when GUS protein was targeted to plastids with the aid of the ibAGP1 transit peptide, GUS levels increased to higher levels in lamina, petiole and cauline leaf compared to those produced by CaMV35S promoter. The enhancing effect of ibAGP1 transit peptide on the accumulation of foreign protein was tissue-specific; accumulation was high in lamina and inflorescence, but low in root and primary inflorescence stem. The transit peptide effect in the leaves was maintained highly regardless of developmental stages of plants. The ibAGP1 promoter and its transit peptide also directed strong GUS gene expression in transiently expressed tobacco leaves. These results suggest that the ibAGP1 promoter and its transit peptide are a strong constitutive foreign gene expression system for transgenesis of dicot plants.  相似文献   

17.
Yang M  Ma QJ  Dang GT  Ma Kt  Chen P  Zhou CY 《Cytotherapy》2005,7(3):273-281
BACKGROUND: Adipose-derived adult stem (ADAS) cells are multipotent cells capable of differentiating into osteoblasts, adipocytes and chondrocytes. The aim of this study was to determine whether BMP-7-expressing ADAS cells would elicit bone formation invitro and in vivo. METHODS: ADAS cells were harvested from Lewis rats and transduced with adenovirus carrying the recombinant human bone morphogenetic protein-7 (Ad-BMP-7) gene. Untransduced cells and cells transduced with adenovirus carrying the enhanced green fluorescence protein (Ad-EGFP) gene served as controls. BMP-7 expression was assessed by RT-PCR, immunofluorescence on day 1, and Western blot on days 4, 8 and 12. Alkaline phosphatase (ALP) activity was assayed on days 2, 4, 6, 8, 10 and 12. Osteocalcin production and bone nodule formation were detected by immunohistochemistry and von Kossa stain on day 12. A total of 1 x 10(6) cells mixed with type I collagen were implanted into the subcutaneous pocket in Lewis rat and subjected to histologic analysis 1, 2 and 4 weeks post-implantation. RESULTS: The Ad-BMP-7-transduced ADAS cells expressed BMP-7 at both mRNA and protein levels. ALP activity was detected in Ad-BMP-7-transduced cells from day 2 to day 12, peaking on day 8. Osteocalcin production and matrix mineralization further confirmed that these cells differentiated into osteoblasts and induced bone formation in vitro. Histologic examination revealed that implantation of BMP-7-expressing ADAS cells could induce new bone formation in vivo. DISCUSSION: ADAS cells would be a promising source of adult autologous stem cells for BMP gene therapy and tissue engineering.  相似文献   

18.
BACKGROUND AND AIMS: Drought causes a decline of root hydraulic conductance, which aside from embolisms, is governed ultimately by aquaporins. Multiple factors probably regulate aquaporin expression, abundance and activity in leaf and root tissues during drought; among these are the leaf transpiration rate, leaf water status, abscisic acid (ABA) and soil water content. Here a study is made of how these factors could influence the response of aquaporin to drought. METHODS: Three plasma membrane intrinsic proteins (PIPs) or aquaporins were cloned from Phaseolus vulgaris plants and their expression was analysed after 4 d of water deprivation and also 1 d after re-watering. The effects of ABA and of methotrexate (MTX), an inhibitor of stomatal opening, on gene expression and protein abundance were also analysed. Protein abundance was examined using antibodies against PIP1 and PIP2 aquaporins. At the same time, root hydraulic conductance (L), transpiration rate, leaf water status and ABA tissue concentration were measured. KEY RESULTS: None of the treatments (drought, ABA or MTX) changed the leaf water status or tissue ABA concentration. The three treatments caused a decline in the transpiration rate and raised PVPIP2;1 gene expression and PIP1 protein abundance in the leaves. In the roots, only the drought treatment raised the expression of the three PIP genes examined, while at the same time diminishing PIP2 protein abundance and L. On the other hand, ABA raised both root PIP1 protein abundance and L. CONCLUSIONS: The rise of PvPIP2;1 gene expression and PIP1 protein abundance in the leaves of P. vulgaris plants subjected to drought was correlated with a decline in the transpiration rate. At the same time, the increase in the expression of the three PIP genes examined caused by drought and the decline of PIP2 protein abundance in the root tissues were not correlated with any of the parameters measured.  相似文献   

19.
20.
Avascular necrosis (AVN) is a disorder of the bone repair process which usually results in femoral head (FH) destruction. Bone morphogenetic proteins (BMPs) are the key proteins regulating bone remodelling and healing. BMPs gene expression levels were analyzed in the normal and necrotic sites of osteonecrotic FHs. Quantitative RT-PCR for BMP-2, -4, -6, -7 genes was performed in bone tissue samples from 47 osteonecrotic FHs. Protein levels of BMP-2, -4, -6 were estimated by Western Blot. Statistical analysis was performed using the Wilcoxon signed rank test. BMP-2 and BMP-6 mRNA levels were higher in the normal than the necrotic site (normal/necrotic: 16.8/6.8 and 1.75/1.64, respectively). On the contrary, BMP-4 mRNA levels were higher in the necrotic (0.75) than the normal (0.62), while BMP-7 mRNA levels were extremely low. At the protein level, BMP-2 continued to have a higher expression in the normal region (normal/necrotic: 0.67/0.64). BMP-4 and -6 were detected at higher levels in the necrotic site (normal/necrotic: 0.51/0.61 for BMP-4, 0.51/0.56 for BMP-6), while BMP-7 was not detectable. Different BMP levels between the normal and necrotic site, as well as discrepancies between the gene and protein expression pattern suggest a different regulation mechanism for BMPs between the two regions of FHs. The understanding of the expression pattern and the correlation of BMPs could lead to a more successful use in the prevention and treatment of AVN.  相似文献   

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