基质细胞衍生因子（SDF-1）及其受体CXCR4对T细胞的生物学效应

趋化因子是目前很多领域研究的热点.SDF-1/CXCR4在造血干/祖细胞动员和归巢及对肿瘤细胞的生存和播散方面均起了重要作用.同时SDF-1/CXCR4对T细胞的分化发育及活化作用也日益被深入研究.本文就SD-1/CXCR4的基本结构及对T细胞的生物学效应作简要综述.     

基质细胞衍生因子(SDF-1)及其受体CXCR4对T细胞的生物学效应

趋化因子是目前很多领域研究的热点。SDF-1/CXCR4在造血干/祖细胞动员和归巢及对肿瘤细胞的生存和播散方面均起了重要作用。同时SDF-1/CXCR4对T细胞的分化发育及活化作用也日益被深入研究。本文就SD-1/CXCR4的基本结构及对T细胞的生物学效应作简要综述。     

基质细胞衍生因子对骨髓干细胞迁移分化的影响

骨髓干细胞具有很强的可塑性，可向多种组织细胞横向分化，是细胞移植的最好种子之一。体内外许多试验证实，某些细胞因子可以影响移植细胞的微环境，进而影响骨髓干细胞的动员、迁移及分化。基质细胞衍生因子属于趋化因子中CXC亚家族，特异性引起表达CXCR4的骨髓干细胞趋化反应，就基质细胞衍生因子对骨髓干细胞动员、迁移及分化的影响作一综述。     

基质细胞衍生因子-1_α/CXCR4/CXCR7轴对骨髓间充质干细胞迁移的影响

目的探讨基质细胞衍生因子-1α(SDF-1α)受体CXCR4、CXCR7在骨髓间充质干细胞(BMSCs)中蛋白和mRNA的表达;及SDF-1α/CXCR4/CXCR7轴对BMSCs迁移作用的可能机制。方法体外培养大鼠BMSCs,流式细胞术鉴定细胞表面抗原CD29、CD44和CD34。应用CXCR4特异性拮抗剂AMD3100及CXCR7中和抗体分别阻断CXCR4及CXCR7,通过Western blotting和RT-PCR分别检测BMSCs蛋白和mRNA的表达变化;Transwell法检测细胞迁移能力。本次实验分为单纯BMSCs组(A)、AMD3100预处理BMSCs组(B)、CXCR7中和抗体预处理BMSCs组(C)及AMD3100+CXCR7中和抗体预处理BMSCs组(D)。结果经鉴定第3代大鼠BMSCs中CD29和CD44均呈阳性表达,而CD34表达阴性。BMSCs中CXCR4、CXCR7蛋白和mRNA均有表达。与A组相比,B组及D组CXCR4及CXCR7蛋白表达明显受到抑制(P0.05),C组只有CXCR7蛋白表达降低(P0.05);各组CXCR4 mRNA和CXCR7 mRNA的表达差异均无显著性。SDF-1α可以诱导BMSCs迁移,与0μg/L组相比,10μg/L组和100μg/L组穿膜细胞数均显著增多(P0.01),与10μg/L组相比,100μg/L组穿膜细胞数亦明显增多(P0.01);AMD3100和CXCR7中和抗体均能抑制BMSCs的迁移作用(P0.05),当两者同时作用时,抑制效应更为显著(P0.05)。结论 BMSCs共表达CXCR4、CXCR7蛋白及mRNA;BMSCs的迁移具有SDF-1α浓度依赖性;SDF-1α/CXCR4/CXCR7轴介导BMSCs的迁移作用,CXCR4受体和CXCR7受体对BMSCs的迁移可能具有协同促进作用。     

基质细胞衍生因子的构效关系研究进展

基质细胞衍生因子1(stromal cell—derived factor-1，SDF—1)及其受体CXCR4广泛表达于多种组织、器官、系统，在人和动物的生理和病理过程中起着重要的作用。弄清SDF-1的结构与效应的关系是阐明诸多生理和病理过程分子机制的关键，在此基础上进行药物设计和优化并应用于临床。本就SDF-1的一级结构和高级结构与功能的关系、SDF-1与多糖的相互作用、SDF-1与CXCR4相互作用机制等方面的最新研究进展作一概述。     

心房快速起搏对犬心肌基质细胞衍生因子-1表达的影响

目的： 研究心房快速起搏犬模型心肌基质细胞衍生因子-1（SDF-1）的表达。方法： 选用成年健康杂种犬13条,随机分为2组：快速起搏组7条,假手术组6条。2组均开胸于右心耳缝植AOO型起搏器，快速起搏组以400 beats/min起搏6周，假手术组不起搏。在实验结束时，应用逆转录-多聚酶链反应（RT-PCR）测定左心耳、左心房的SDF-1 mRNA表达水平,同时进行左心房的病理及免疫组化分析。结果：快速起搏组犬的左心耳、左心房的SDF-1 mRNA表达水平明显高于假手术组（P<0.05）,分别增高18.5%和22.4%；病理结果示快速起搏组犬左心房心肌细胞变性；免疫组化测定显示快速起搏组犬的左心房SDF-1表达明显多于假手术组，主要分布在心内膜下的心肌细胞内。结论： 心房快速起搏可引起犬心肌组织SDF-1 mRNA及蛋白表达水平增高。SDF-1可能参与心房损伤时心肌组织的修复过程，并对心肌的重构起一定作用。     

基质细胞衍生因子的构效关系研究进展

基质细胞衍生因子1(stromalcell鄄derivedfactor鄄1,SDF鄄1)及其受体CXCR4广泛表达于多种组织、器官、系统,在人和动物的生理和病理过程中起着重要的作用。弄清SDF鄄1的结构与效应的关系是阐明诸多生理和病理过程分子机制的关键,在此基础上进行药物设计和优化并应用于临床。本文就SDF鄄1的一级结构和高级结构与功能的关系、SDF鄄1与多糖的相互作用、SDF鄄1与CXCR4相互作用机制等方面的最新研究进展作一概述。     

长期培养对基质细胞衍生因子1/CXCR4介导间充质干细胞迁移的影响**◆

背景：人外源性的间充质干细胞能特异性地向损伤部位迁移,参与多种组织的损伤修复,然而其定向迁移的机制尚不清楚。 目的：观察基质细胞衍生因子1对骨髓间充质干细胞定向迁移的影响。 方法：体外培养不同个体的骨髓间充质干细胞,培养至第10代,检测细胞衰老状态。RT-PCR和细胞免疫荧光检测骨髓间充质干细胞CXCR4受体表达情况;体外迁移体系(Transwell)检测基质细胞衍生因子1对骨髓间充质干细胞迁移的影响。 结果与结论：骨髓间充质干细胞在体外长期增殖后生长逐渐缓慢,在第6,7代衰老细胞增多,其CXCR4受体表达减少。基质细胞衍生因子1对骨髓间充质干细胞的趋化作用呈剂量依赖性。 关键词：趋化作用;骨髓间充质干细胞;细胞衰老;基质细胞衍生因子1;CXCR4受体 doi:10.3969/j.issn.1673-8225.2012.01.013      

基质细胞衍生因子1趋化神经干细胞迁移的体外效应

背景：神经干细胞的迁移在神经系统的发育和损伤修复中起着至关重要的作用,近来研究表明趋化因子参与神经干细胞的迁移,但关于其迁移机制目前尚不清楚。 目的：观察体外条件下基质细胞衍生因子1对胎鼠海马神经干细胞的趋化迁移作用。 方法：通过无血清法体外分离、培养及鉴定胎鼠海马神经干细胞;细胞免疫荧光及RT-PCR检测其CXCR4是否表达;观察不同浓度基质细胞衍生因子1对神经干细胞的趋化迁移作用,中和CXCR4受体以验证基质细胞衍生因子1趋化迁移作用的特异性。 结果与结论：胎鼠海马来源神经干细胞表达趋化因子受体CXCR4,呈阳性。RT-PCR琼脂糖凝胶电泳后出现643 bp特异性扩增条带。体外条件下基质细胞衍生因子1趋化迁移随浓度而增强,500 μg/L为最佳趋化浓度。加入抗CXCR4多克隆抗体中和后,神经干细胞迁移较基质细胞衍生因子1组明显减少,与对照组比较,差异无显著性意义(P > 0.05),提示抗CXCR4多克隆抗体可阻断趋化迁移作用。     

SDF1/CXCR4信号轴在退变髓核细胞诱导血管新生中的作用

目的探讨髓核细胞(NPCs)分泌的基质细胞衍生因子1(SDF1)对退变椎间盘血管化的影响及相关分子机制。方法培养退变椎间盘NPCs,病毒转染分别上调和下调SDF1表达,根据SDF1表达情况分为DOWN、D和UP 3组;培养人脐静脉内皮细胞(HUVECs),将不同组NPCs或其条件培养基与HUVECs进行共培养,期间还在共培养体系中加入CXCR4受体抑制剂AMD3100,CCK8检测、细胞迁移实验、管腔形成实验观察HUVECs成血管能力。结果各组NPCs构建成功;与HUVECs共培养后,利用CCK8法、细胞迁移实验、管腔形成实验检测结果显示随SDF1表达升高,内皮细胞活力、迁移能力、管腔形成能力也随之升高(P0.05);当加入AMD3100后,内皮细胞活力、迁移能力、管腔形成能力在D组和UP组均受到显著抑制(P0.05)。结论人退变椎间盘NPCs表达SDF1。SDF1/CXCR4信号轴影响内皮细胞活力、迁移能力和成管能力,并在退变椎间盘血管化中发挥重要作用。     

自体骨髓单个核细胞移植治疗缺血性心脏病临床疗效观察

目的 研究经冠状动脉内移植骨髓单个核细胞治疗缺血性心力衰竭的可行性和疗效。方法 缺血性心力衰竭病人20例，抽取骨髓用密度梯度离心法分离出骨髓单个核细胞，经外周动脉穿刺插管，将自体骨髓单个核细胞注入冠脉。比较病人的临床症状，NYHY分级和治疗后由于心血管事件造成的再住院情况。结果 病人自觉症状改善，心功能提高，ECT检查显示在缺血区有血管新生。结论 自体骨髓单个核细胞移植是安全有效的。     

Morphological and molecular analysis of angiogenesis after intramyocardial transplantation of autologous bone marrow mononuclear cells

We studied the peculiarities of angiogenesis in the postinfarction period after transmyocardial laser revascularization and intramyocardial implantation of mononuclear bone marrow cells into the pericicatrical zone of the left ventricular myocardium in dogs. Morphological manifestation of angiogenesis in the myocardium after application of laser and cell technologies are angiomatosis, formation of large thin-wall vessels and sinusoids. The angiogenic effect of implanted mononuclear bone marrow cells is determined by high content (43-47%) of CD31⁺ cells in both adherent and nonadherent fractions. More pronounced angiogenic potential of nonadherent cells is determined by intensive expression of cytokine VEGF-B and D mRNA essential for arterial vessels growth. Immunohistochemical studies showed that about 90% cells of the nonadherent fraction are endothelial precursors expressing endothelial cell markers isolectin B₄ and VEGF-R2. It was found that the use of adherent mononuclear bone marrow cells during the postinfarction period induces ossification of the epicardium and subepicardial myocardium layer, formation of cartilage plates, and focal calcification. Implantation of nonadherent mononuclear bone marrow cells into transmyocardial laser channels did not induce ectopic ossification of the myocardium.     

Cardiac contractility after transplantation of autologous mononuclear bone marrow cells in patients with myocardial infarction

Autologous bone marrow mononuclear cells were transplanted by intracoronary infusion to patients with myocardial infarction after recovery of coronary perfusion. Controls received traditional therapy alone. Echocardiography was carried out before and 3 and 6 months after cell therapy. Cell transplantation did not appreciably improved left-ventricular contractility in comparison with the control group. In none patient cell therapy provoked malignant ventricular arrhythmias. Intracoronary infusion of bone marrow mononuclear cells in patients with myocardial infarction did not improve cardiac contractility and did not aggravate the course of the disease. __________ Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 1, pp. 15–20, January, 2006     

Neovascularization and bone regeneration by implantation of autologous bone marrow mononuclear cells

We examined whether transplantation of autologous bone marrow mononuclear cells (BM-MNCs) can augment neovascularization and bone regeneration of bone marrow in femoral bone defects of rabbits. Gelatin microspheres containing basic fibroblast growth factor (bFGF) were prepared for the controlled release of bFGF. To evaluate the in vivo effect of implanted BM-MNCs, we created bone defects in the rabbit medial femoral condyle, and implanted into them 5 x 10(6) fluorescent-labeled autologous BM-MNCs together with gelatin microspheres containing 10 microg bFGF on an atelocollagen gel scaffold. The four experimental groups, which were Atelocollagen gel (Col), Col + 5 x 10(6) BM-MNCs, Col + 10 microg bFGF, and Col + 5 x 10(6) BM-MNCs + 10 microg bFGF, were implanted into the sites of the prepared defects using Atelocollagen gel as a scaffold. The autologous BM-MNCs expressed CD31, an endothelial lineage cell marker, and induced efficient neovascularization at the implanted site 2 weeks after implantation. Capillary density in Col + BM-MNCs + bFGF was significantly large compared with other groups. This combination also enhanced regeneration of the bone defect after 8 weeks to a significantly greater extent than either BM-MNCs or bFGF on their own. In summary, these findings demonstrate that a combination of BM-MNCs and bFGF gelatin hydrogel enhance the neovascularization and the osteoinductive ability, resulting in bone regeneration.     

自体骨髓单个核细胞肝内移植治疗失代偿期肝硬化

背景：大量研究发现,骨髓间充质干细胞是肝干细胞的主要肝外来源,骨髓间充质干细胞在特定环境下可分化为肝组织和肝细胞从而参加肝脏的修复与重建。 目的：观察自体骨髓单个核细胞移植治疗失代偿期肝硬化的效果及安全性。 方法：选择肝硬化失代偿期患者20例,在无菌条件下由髂后上棘抽取骨髓200 mL,在体外分离纯化骨髓单个核细胞并制成10 mL细胞悬液,经肝动脉将细胞悬液移植入肝脏,分别在移植后第2、4、8、12周复查肝脏功能,观察实验室指标,临床症状及不良反应。 结果与结论：移植后2周内所有血清学指标均无显著改变,移植后4周白蛋白由(27.05±5.23) g/L升到(30.02±5.02) g/L,纤维蛋白原由(1.55±0.53) g/L升到(2.55±0.53) g/L,凝血酶原时间由(24.05±5.23) s降到 (17.05±5.13) s,胆碱酯酶和甲胎蛋白也有不同程度上升,氨基转移酶、总胆红素水平在移植前后无显著性变化。单个核细胞移植后食欲改善、体力好转10例,腹胀减轻6例,腹水减少7例,下肢浮肿减轻4例。全部患者在移植中未发生严重并发症,移植近期无不良反应出现。     

Reparation of the myocardium after transplantation of mononuclear bone marrow cells

Safety and efficiency of intracoronary transventricular transplantation of autologous mononuclear bone marrow cells in rats with postinfarction cardiosclerosis were studied. The cells migrated to the damaged area and were detected only in the cicatricial tissue; they have fibroblast-like phenotype and some of them were stained with Fapα (marker of reactive fibroblasts). More active proliferation of non-muscular cells and formation and maturation of collagen fibers in the cicatricial tissue were observed after transplantation of mononuclear cells. This led to thickening of the cicatricial wall, but the size of the scar and index of dilatation of the left ventricle remained unchanged. The number and volume density of newly formed blood vessels in the damaged area increased after transplantation, but no labeled cells were seen in the vascular walls. It can be hypothesized that stimulation of neoangiogenesis is mediated by paracrine mechanisms, which also explains improvement of global contractility of the left ventricle (increased contractility index in functional tests). Thus, transplantation of mononuclear bone marrow cells leads to thickening and strengthening of the cicatricial wall, stimulates angiogenesis, and improves global myocardial contractility. However, no morphological signs of reverse remodeling of the left-ventricular myocardium were revealed.     

SDF-1/CXCR4增强骨髓间质干细胞的造血支持作用

目的探讨基质细胞衍生因子-1(SDF-1)/CXCR4在骨髓间质干细胞(MSCs)支持CD34 造血干/祖细胞(HSPCs)扩增中的作用。方法在长期培养基(LTC)中,以大鼠骨髓MSCs作为饲养层体外扩增骨髓CD34 细胞,每周分别加入SDF-1、SDF-1抗体或CXCR4抗体至5周。计算CD34 细胞数和集落形成细胞(CFC)数,以评价造血支持功能。为评估SDF-1/CXCR4对CD34 细胞增殖周期的影响,进行了杀伤试验以计算增殖指数。流式细胞术检测MSCs和CD34 细胞中SDF-1与CXCR4的表达;ELISA检测MSCs和CD34 细胞培养基中SDF-1的含量。结果CD34 细胞数、CFC数和增殖指数在加入SDF-1后明显增加(P<0.01),加入SDF-1抗体或CXCR4抗体后明显减少(分别为P<0.05,P<0.01)。CD34 细胞表面表达CXCR4,MSCs则不表达;MSCs细胞内表达SDF-1,而CD34 细胞不表达。在MSCs培养基中检测到SDF-1,在CD34 细胞培养基中未发现。结论SDF-1/CXCR4在骨髓MSCs支持HSPCs扩增中起重要作用。     

自体骨髓单个核细胞移植治疗大鼠后肢缺血

目的　探讨自体骨髓单个核细胞（BM-MNC）移植用于大鼠缺血后肢的治疗后实现血管再生的能力。方法　建立大鼠后肢缺血动物模型,将取于自体的BM-MNC制成细胞悬液注射于缺血部位,分别在移植后2和30d时行动脉造影,用免疫组化方法检测内皮祖细胞（EPC）,毛细血管密度以及测定血管内皮生长因子(VEGF)的表达。结果　缺血肌组织中的EPC含量增高（P<0.01）。BM-MNC移植组在移植早期VEGF表达显著增高（P<0.01）。细胞移植后30dBM-MNC移植组毛细血管密度明显高于其他组（P<0.01）,血管造影可见侧支循环建立。结论　自体骨髓单个核细胞移植于大鼠后肢缺血区能促进血管新生,改善侧支循环,可望成为一种简单有效的治疗下肢缺血的方法。     

Scedosporium apiospermum pneumonia after autologous bone marrow transplantation

Although opportunistic infections after bone marrow transplantation (BMT) are very common, only five cases ofPseudallescheria boydii infection have been reported in the literature, two of which were autopsy findings. A case ofScedosporium apiospermum infection after BMT, treated initially with amphotericin B (total dose of 2.5 g) and then with itraconazole (for 25 days), is reported here. When the patient failed to improve,Scedosporium apiospermum pneumonia was diagnosed and therapy was changed. The patient was treated successfully with miconazole (600 mg/8h for 32 days) and ketoconazole (200 mg/8h for 7 days) plus surgery.