RGD三肽的合成与生物活性检测

在生物材料表面上先接枝RGD,然后再在其上种植和培养内皮细胞,这是目前改善心血管材料血液相容性最好的方法之一。采用液相合成法合成了RGD 三肽并对其进行了生物活性检测,证实用液相合成的 RGD 短肽具有促进材料与内皮细胞粘附的生物活性。     

化学-酶法合成RGD三肽

采用化学法和酶法相结合的合成策略,合成具有高生物活性肽RGD.先采用DCC合成GD二肽(HCl*Gly-Asp(oBzl)2),然后利用胰蛋白酶(trypsin)催化合成 RGD三肽(Z-Arg-Gly Asp(oBzl)2).实验结果表明,其中GD二肽在反应10 h,两底物G与D的摩尔比为1.5∶1时,得率为77.7%;RGD三肽在微水-1,4-丁二醇反应体系中得率为41.9%.     

掺Mg与接枝RGD的HAnps载体系统对MG63细胞胞吞功能的影响

采用水热合成法制备HAnps和掺镁HAnps(Mg-HAnps),通过硅烷化联合碳二亚胺法处理HAnps和Mg-HAnps后接枝黏附肽(RGD),以香豆素6(coumarin-6)为荧光探针标记HAnps,Mg-HAnps,RGD-HAnps和RGD-Mg-HAnps,并将其与人成骨肉瘤MG63细胞株(MG63)细胞共培养,研究掺Mg与接枝RGD的HAnps对MG63细胞胞吞作用的影响。研究结果表明:RGD-Mg-HAnps无细胞毒性,掺Mg与接枝RGD有利于MG63细胞胞吞HAnps颗粒,并有协同作用,RGD-Mg-HAnps具有介导胞吞的作用。     

细胞粘附肽Arg-Gly-Asp的合成与生物活性检测

采用固相多肽合成法合成Ag-G ly-Asp(RGD)三肽,以天门冬氨酸计RGD三肽收率为75%,采用MTT法进行合成肽RGD抑制人纤维肉瘤细胞HT1080转移机理方面的研究,证实了人工合成的RGD肽能抑制人纤维肉瘤细胞HT1080与纤维连接蛋白(FN)的粘附。     

细胞粘附肽Arg-Gly-Asp的合成与生物活性检测

采用固相多肽合成法合成Ag-Gly-Asp(RGD)三肽,以天门冬氨酸计RGD三肽收率为75%,采用MTT法进行合成肽RGD抑制人纤维肉瘤细胞HT1080转移机理方面的研究,证实了人工合成的RGD肽能抑制人纤维肉瘤细胞HT1080与纤维连接蛋白(FN)的粘附.     

RGD对人纤维肉瘤细胞HT1080增殖、黏附及转移影响的研究

利用合成的RGD(Arg-Gly-Asp)三肽研究了其抑制人纤维肉瘤细胞HT1080的增殖、黏附及转移作用.采用MTT法检测了不同浓度的RGD对HT1080细胞增殖及黏附纤维粘连蛋白(fibronectin)能力的影响;采用细胞划痕法研究了RGD对HT1080细胞在纤维粘连蛋白中迁移能力的影响.结果表明,合成的RGD能抑制HT1080细胞的增殖,并具有抗肿瘤细胞黏附、抗转移的活性.     

高岭土-丙烯酰胺系超吸水性复合材料表征

应用X－射线粉晶衍射（XRD）、红外吸收光谱（IR）的方法，对高岭土－淀粉接枝共聚丙烯酰胺超吸水性复合材料进行表征。结果表明，在超吸水性复合材料的制备中，高岭土粉体的结构依然保存，分散度提高。同时，高岭土粉体表面的羟基、淀粉上的羟基与丙烯酰胺单体等，发生了接枝共聚反应。     

RGD多肽接枝聚(乳酸-羟基乙酸-L-赖氨酸)的制备与表征

以3S-[4-（苄氧羰基氨基）丁基]-吗啉-2,5-二酮和丙交酯为起始原料,制备一种新型的RGD多肽接枝聚（乳酸-羟基乙酸-L-赖氨酸）共聚物（PRGD）。采用核磁共振氢谱、氨基酸分析、接触角测试、MTT实验和环境扫描电镜对其结构和性能进行表征。研究结果表明：RGD接枝量为6．9～14.3μmol／g;PRGD膜和聚乳酸（PLA）膜的水接触角分别为43．63°和62．45°;PRGD膜表面黏附的嗅鞘细胞较PLA组活性高,细胞密度大,生长状态好。PRGD较PLA具有更好的亲水性和神经细胞亲和性,有望成为一种理想的神经修复材料。     

4种含RGD的短肽对细胞粘附作用的影响

从两方面对比研究了4种含Arg-Gly-Asp(RGD)的细胞粘附肽RGD,RGDS,RGD-(NH₂)₂即RGE-NH_{2和RGDS-NH2对细胞粘附的影响. 一方面研究了固定在聚乙交酯丙交酯共聚物（PLGA）膜上的4种细胞粘附肽通过其细胞定向作用对大肠癌细胞（HCT-8）和人成骨肉瘤细胞（OS732）细胞粘附性 的促进作用; 另一方面研究了外源性细胞粘附肽对HCT-8和OS732在纤维连接蛋白（FN）上粘附的竞争性抑制作用. 结果发现,除了RGD-(NH2)2,其他3种肽都具有明显抑制细胞粘附的作用且具有一定的浓度依赖关系,其中以RGDS和RGDS-NH2的能力最强,RGD稍弱. 对于HCT-8细胞,3种肽的最大抑制率分别为53.3%, 56.3%,37.5%;而对OS732细胞,3种肽的最大抑制率分别为50.9%,49.8%,34%.}     

三氮唑羧酸酯类席夫碱的合成及生物活性

从５－氨基－１Ｈ－１，２，４－三氮唑－３－羧酸出发，合成了７种５－氨基－１Ｈ－１，２，４－三氮唑－３－羧酸甲酯类席夫碱，对其产率，物理性持作了首次报道，采用ＵＶ，ＩＲ，＾１ＨＮＭＲ和元素分析等对化合物结构进行了初步表征，实验室生物活性测试表明此类化合物具有较好的生物活性。     

ICAM, an adhesion ligand of LFA-1, is homologous to the neural cell adhesion molecule NCAM

Antigen-specific cell contacts in the immune system are strengthened by antigen-nonspecific interactions, mediated in part by lymphocyte-function associated (LFA) antigens. The LFA-1 antigen is widely expressed on cells of haematopoietic origin and is a major receptor of T cells, B cells and granulocytes. LFA-1 mediates the leukocyte adhesion reactions underlying cytolytic conjugate formation, helper T-cell interactions, and antibody-dependent killing by natural killer cells and granulocytes. Recently, ICAM-1 (intercellular adhesion molecule-1) has been defined as a ligand for LFA-1. Monoclonal antibodies to ICAM-1 block T lymphocyte adhesion to fibroblasts and endothelial cells and disrupt the interaction between cytotoxic T cells and target cells. In addition, purified ICAM-1 reconstituted into artificial membranes binds LFA-1+ cells. ICAM-1 is found on leukocytes, fibroblasts, epithelial cells and endothelial cells and its expression is regulated by inflammatory cytokines. LFA-1 has been placed in the integrin family of cell surface receptors by virtue of the high sequence similarity between the LFA-1 and integrin beta chains. The adhesion ligands of the integrin family are glycoproteins bearing the Arg-Gly-Asp (RGD) sequence motif, for example, fibronectin, fibrinogen, vitronectin and von Willebrand factor. Here we show that a complementary DNA clone ICAM-1 contains no RGD motifs, but instead is homologous to the neural cell adhesion molecule NCAM.     

清除血浆中LDL用生物活性短肽SDE的合成研究

高脂血症是引起心脑血管疾病的主要原因之一。Ser-Asp-Glu 三肽(SDE)在对血浆中低密度脂蛋白(LDL)的识别及特异性吸附起关键性作用。本实验采用 DCC/HOBt 液相法,合成三肽(SDE)。重点研究反应温度、反应时间和原料配比对 SDE 产率的影响。结果表明本实验成功合成出SDE 三肽。     

紫外辐照在PET膜表面接枝氨基酸的研究

生物材料的表面性能是影响材料血液相容性的重要原因,生物材料的表面改性对提高其血上容性有重要意义,目前国内外公认的材料表面内皮细胞化是提高生物材料血液相容性最为理想的途径。     

丝素包埋SPA成膜的理化性能和生物效应研究

研究将丝素溶液包埋金黄色葡萄球菌A蛋白(staphylococcal protein A)(简称A蛋白或SPA ),制成不溶性SPA丝素膜.用粘附生长因子RGD(序列为GLY-ARG-GLY-ASP-SER-PRO-L YS)的抗体IgG结合IgG到该丝素膜的表面,再将RGD结合到不溶性SPA丝素膜表面RGD抗体IgG 上,制备成RGD-IgG-SPA丝素膜.用酶联免疫吸附试验(简称ELISA)方法,证明RGD-IgG -SPA丝素膜是稳定的.种植培养血管内皮细胞(vascular endothelial cell,简称EC细胞) 于该改性丝素膜的表面,用四甲基偶氮唑盐比色方法(简称MTT法)检测细胞的生长,证明改性丝素膜能有效地促进EC细胞的生长.     

RGD peptides induce apoptosis by direct caspase-3 activation

Synthetic peptides containing the arginine-glycine-aspartate (RGD) motif have been used extensively as inhibitors of integrin-ligand interactions in studies of cell adhesion, migration, growth and differentiation, because the RGD motif is an integrin-recognition motif found in many ligands. Here we report that RGD-containing peptides are able to directly induce apoptosis without any requirement for integrin-mediated cell clustering or signals. We show that RGD-containing peptides enter cells and directly induce autoprocessing and enzymatic activity of procaspase-3, a pro-apoptotic protein. Using the breast carcinoma cell line MCF-7, which has a functional deletion of the caspase-3 gene, we confirm that caspase-3 is required for RGD-mediated cell death. In addition to an RGD motif, pro-caspase-3 also contains a potential RGD-binding motif, aspartate-aspartate-methionine (DDM), near the site of processing to produce the p12 and p17 subunits. On the basis of the ability of RGD-DDX interactions to trigger integrin activation, we suggest that RGD peptides induce apoptosis by triggering conformational changes that promote pro-caspase-3 autoprocessing and activation. These findings provide an alternative molecular explanation for the potent proapoptotic properties of RGD peptides in models of angiogenesis, inflammation and cancer metastasis.     

Cbz全保护RGD-OH的合成方法研究

采用碳端到氮端依次连接策略,以天冬氨酸、甘氨酸、精氨酸为起始原料,以HBTU为缩合剂,Asp(OBzl)-OAllyl顺次分别与Boc-甘氨酸和Cbz-Arg(Cbz)2偶联生成碳端和氮端保护的RGD。进一步在N-甲基吗啉存在下以Pd(PPh3)4催化脱去烯丙基,得到目标产物。中间产物和目标产物的分离、提纯采用重结晶和柱层析法,应用1 H NMR,13C NMR和MS等对其结构进行了表征。     

Molecular imaging of tumor angiogenesis using RGD-labeled iron oxide nanoparticles

Integrin is often significantly up­regulated in activated endothelial cells during tumor angiogenesis. The arginine-glycine-aspartic acid (RGD) peptide sequence is a specific recognition motif to α_νβ₃ integrin. In this study, a RGD labeled, Poly lactic acid (PLA) coated ultrasmall paramagnetic iron oxide (USPIO) (referred to as RGD-PLA-USPIO) were developed and the ability to detect tumor angiogenesis was investigated in vitro and in vivo. Increased uptake of RGD-PLA-USPIO by human umbilical vein endothelial cells (HUVECs) was detected by Prussian blue stain and transmission electronic microscopy (TEM). Pronounced signal decrease in T₂^*-weighted magnetic resonance image (MRI) and heterogeneous arrangement of neovasculature of tumor tissue were clearly identified in Vx-2 tumor model. The MR signal of contralateral muscle only could be seen a slight background change after either RGD-PLA-USPIO or PLA-USPIO injection. These studies demonstrate the efficiency of RGD-PLA-USPIO to visualize α_νβ₃ integrin in activated tumor endothelial cells and its potential for detecting and monitoring tumor vasculature change after therapy.