腺病毒介导人VEGF_(165)和ANG-1双基因转染大鼠BMSCs及对其增殖的影响

[目的]探讨携带人血管内皮细胞生长因子165(VEGF165)和血管生成素-1(ANG-1)双基因的腺病毒表达载体pAd-VIA转染大鼠骨髓基质干细胞(BMSCs)的体外表达及对BMSCs增殖的影响。[方法]将本室构建的编码人VEGF165和ANG-1双基因的腺病毒质粒pAd-VIA在QBI-293A细胞内进行包装和扩增,体外转染大鼠BMSCs,通过绿色荧光蛋白(GFP)表达、Western-blotting、酶联免疫吸附分析(ELISA)方法检测外源基因的表达,利用MTT法检测MOI(multiplicity of infection)=50、100、200、400 pfu/cell不同浓度腺病毒转染后对BMSCs增殖活性的影响。[结果]重组腺病毒质粒pAd-VIA在QBI-293A细胞内成功包装和扩增,体外转染BMSCs后,有大量的GFP表达,Western-blotting检测显示,转染组与VEGF165和ANG-1抗体结合,在45 KD和14.4 KD左右出现印迹条带;ELISA结果显示:转染组第1、2、3 d,上清中的VEGF165浓度和ANG-1浓度持续增加,未转染组均未检测到外源基因的表达,差异...     

腺病毒介导入VEGF165和ANG-1双基因转染大鼠BMSCs及对其增殖的影响

[目的]探讨携带人血管内皮细胞生长因子165(VEGF165)和血管生成素-1(ANG-1)双基因的腺病毒表达载体pAd-VIA转染大鼠骨髓基质干细胞(BMSCs)的体外表达及对BMSCs增殖的影响.[方法]将本室构建的编码人VEGF165和ANG-1双基因的腺病毒质粒pAd-VIA在QBI-293A细胞内进行包装和扩增,体外转染大鼠BMSCs,通过绿色荧光蛋白(GFP)表达、Western-blotting、酶联免疫吸附分析(ELISA)方法检测外源基因的表达,利用MTT法检测MOI(multiplicity of infection)=50、100、200、400 pfu/cell不同浓度腺病毒转染后对BMSCs增殖活性的影响.[结果]重组腺病毒质粒pAd-VIA在QBI-293A细胞内成功包装和扩增,体外转染BMSCs后,有大量的GFP 表达,Western-blotting检测显示,转染组与VEGF165和ANG-1抗体结合,在45 KD和14.4 KD左右出现印迹条带;ELISA结果显示:转染组第1、2、3 d,上清中的VEGF165浓度和ANG-1浓度持续增加,未转染组均未检测到外源基因的表达,差异有统计学意义(P<0.01);不同浓度的腺病毒载体转染BMSCs后,促进细胞增殖,细胞生长曲线上移,在1、3、5、7 d转染组的OD值均大于未转染组,差异有统计学意义(P<0.01),第9 d,细胞进入平台期,转染组和未转染组的OD值差异无统计学意义(P>0.05).[结论]重组腺病毒pAd-VIA转染大鼠BMSCs后,外源基因在体外得到有效表达,同时在观察时间内可以促进大鼠BMSCs的增殖.     

联合转染人VEGF165和ANG-1双基因对大鼠骨髓基质干细胞成骨分化的影响

[目的]研究联合转染入血管内皮细胞生长因子165(VEGF165)和血管紧张素-1(ANG-1)双基因对大鼠骨髓基质干细胞(BMSCs)成骨分化潜能的影响.[方法]将本室构建的编码人VEGF165和ANG-1双基因腺病毒质粒pAd-VIA在QBI-293A细胞内进行包装和扩增,获得腺病毒载体pAd-VIA,将其体外转染大鼠BMSCs,应用诱导培养液定向诱导向成骨细胞分化.实验分为4组:A.腺病毒载体pAd-VIA转染BMSCs并诱导组;B.BMSCs诱导组;C.腺病毒载体pAd-VIA转染BMSCs组;D.单纯BMSCs组.通过碱性磷酸酶(ALP)染色和活性的测定、Ⅰ型胶原免疫荧光染色、茜素红染色来评价联合转染双基因对BMSCs成骨分化潜能的影响.[结果]重组腺病毒质粒pAdVIA在QBI-293A细胞内成功包装和扩增,有大量的绿色荧光蛋白表达,体外转染大鼠BMSCs后,A、B组的ALP染色、Ⅰ型胶原免疫荧光染色、茜素红染色为阳性,而C组和D组为阴性.ALP活性表达具有时间相关性,3 d开始表达,7 d到达高峰,在7、14 d,A、B组与C、D组之间ALP表达均具有统计学差异(P＜0.01),A组和B组之间,在各个时间点均无统计学差异(P＞0.05).[结论]腺病毒载体pAd-VIA转染大鼠BMSCs后,未明显影响其体外成骨分化潜能.     

共表达腺病毒载体构建鉴定及其转染骨髓间充质干细胞观察

目的：构建并鉴定绿色荧光蛋白标记的人骨形态发生蛋白（bone morphogenetic protein,BMP）2和血管内皮生长因子（vascular endotheliel growth factor,VEGF）165双基因共表达的重组腺病毒,为研究该双基因对骨髓间充质干细胞成骨方向诱导和体内骨缺损修复作用奠定基础。方法：从cDNA文库中PCR扩增BMP2和VEGF165目的基因,并将其插入腺病毒穿梭质粒pAd-MCMV-GFP的多克隆位点,将构建的重组穿梭质粒pAd-MCMV-BMP2-VEGF165和腺病毒辅助质粒pBHGloxΔE1,3Cre共转染细胞HEK293细胞内,经历腺病毒基因重组及出毒后收集细胞,多次冻融离心获取病毒溶液（Ad-BMP2-VEGF165）。进一步纯化并测定病毒滴度,随后通过转染兔骨髓间充质干细胞并检测BMP2和VEGF165双目的基因表达和倒置荧光显微镜下观察绿色荧光蛋白表达。结果：基因测序、菌落PCR、Western blotting、Real-time PCR、绿色荧光蛋白表达均表明载体Ad-BMP2-VEGF165构建成功,可稳定转染骨髓间充质干细胞内并稳定表达,经测定腺病毒载体滴度达到1×10¹⁰ PFU/ml。结论：携带人BMP2和VEGF165双基因共表达重组腺病毒载体构建成功,并能获得高滴度的病毒感染液。     

ANG-1基因重组腺病毒载体的构建及其转染骨髓问充质干细胞的实验研究

目的构建携带大鼠血管生成素．1（angiopoietin-1,ANG．1）基因的重组腺病毒载体,并检测其转染对大鼠骨髓间充质干细胞（bone marrow mesenchymal stemcells,BMSCs）活力的影响。方法RT-PCR法获取大鼠ANG-1基因并亚克隆至穿梭质粒pAdTrack-CMV,经测序无误后与骨架质粒pAdEasy-1在BJ5183中同源重组。重组质粒pAdEasy-1-ANG-1经鉴定后转染293细胞进行病毒包装扩增。体外转染BMSCs,检测转染后BMSCs中ANG-1的表达。MTT法评估常氧及缺氧环境下ANG-1对BMSCs的影响。结果重组腺病毒载体pAdEasy-1-ANG-1经测序及酶切鉴定正确;BMSCs经转染ANG-1基因后表达ANG-1。MTT法检测提示常氧及缺氧条件下转染前后BMSCs活性的差异均无统计学意义（缺氧组与缺氧下转染组相比,P〉0-05;常氧组与常氧下转染组相比,P〉0-05）。结论成功构建大鼠ANG-1基因重组腺病毒载体,且其转染在体外不影响BMSCs的活性。     

人血管内皮细胞生长因子重组腺病毒载体的构建、鉴定及在骨髓间充质干细胞的表达

目的构建人血管内皮细胞生长因子(VEGF)165腺病毒表达载体,体外转染大鼠骨髓间充质干细胞研究其相关特性。方法利用细菌内同源重组技术快速构建Ad-VEGF165腺病毒重组质粒,经酶切及测序鉴定正确后转染人胚肾细胞HEK293包装成为重组Ad-VEGF165腺病毒,并进行电镜观察及滴度测定。感染大鼠骨髓间充质干细胞观察VEGF165基因的转录和表达。结果酶切鉴定及基因测序证实重组腺病毒质粒含有hVEGF165cDNA,电镜显示包装细胞中有大量病毒颗粒存在,测定包装的病毒滴度为6.3×1010TCID50/ml。逆转录-聚合酶链反应(RT-PCR)、免疫组织化学及免疫印迹检测骨髓间充质干细胞内有VEGF165的转录和表达。结论构建的VEGF腺病毒表达载体可有效感染骨髓间充质干细胞,并在体外高效表达,为将表达VEGF基因的骨髓间充质干细胞用于基因治疗提供了实验依据。     

脂质体介导血管内皮生长因子基因片段在犬骨髓基质干细胞中的表达及其影响

目的 探讨脂质体介导的重组血管内皮生长因子165(vascularendothelial growthfactor 165,VEGF165)基因转染后的犬骨髓基质干细胞分泌血管内皮生长因子(vascular endothelialgrowth factor,VEGF)的能力及细胞增殖能力的改变,为改善骨缺血坏死的干细胞治疗方法提供基础.方法 实验分VEGF165基因转染组与空白对照组,从成年犬的胫骨抽取骨髓,体外分离纯化,贴壁培养骨髓基质干细胞,传代培养后以脂质体法将携带荧光蛋白VEGF165基因的穿梭质粒转入实验组骨髓基质干细胞中,在荧光显微镜视下观察转染效率,并通过ELISA法分别检测转染后1、3、5、7、9、11、13 d实验组与对照组培养液中VEGF的含量,对结果进行统计分析.MTT法检测转染细胞的增殖能力,绘制增殖曲线.结果 重组人血管内皮生长因子165(recombinate the human being vasular endotheial growth factor165,hVEGF165)基因通过脂质体能够成功转染犬骨髓基质干细胞并分泌VEGF.h-VEGF165基因转染组培养液中VEGF蛋白含有量较未转染组增高,ELISA结果显示转染后上清液中第2天即可出现VEGF浓度的增高,到第7天时达到分泌高峰,筛选后的细胞上清液中VEGF浓度稳定在300ng/L左右,与对照组相比升高约10倍,其差别具有显著的统计学意义.骨髓基质干细胞转染基因后,细胞的增殖能力同对照组相比无明显差别.结论 采用基因转染技术可将hVEGF165基因转染到犬骨髓间充质干细胞中并可有效表达VEGF. hVEGF165基因转染犬骨髓基质干细胞对细胞的增殖能力无明显影响.     

携带人血管内皮细胞生长因子基因细菌内重组腺病毒转染兔骨髓基质干细胞的表达

目的应用高效细菌内同源重组系统pAd-Easy,制备含人血管内皮细胞生长因子121 (VEGF121)基因的重组腺病毒,感染兔骨髓基质干细胞(BMSCs),并检测外源基因的表达,为其进一步应用于血管化组织工程骨组织的构建打下基础。方法自pCDNA-VEGF121质粒中切取VEGF121基因,将VEGF基因克隆到穿梭质粒,在BJ5183细菌内重组,在293细胞中构建携带VEGF基因的重组腺病毒,将腺病毒感染BMSCs,利用ELISA、免疫组织化学的方法检测VEGF121在BMSCs中的表达。结果通过pAd-Easy系统成功构建高滴度的携带VEGF121基因重组腺病毒pAd-VEGF,ELISA检测显示经转染的BMSCs中VEGF121的表达均明显增强,随着感染病毒MOI值的增高,VEGF121的表达量相应增高,差异有统计学意义(P<0．01),免疫组化法显示经基因转染的BMSCs中有强阳性信号,未转染组出现阴性结果。结论Ad-VEGF转染BMSCs后能稳定提高VEGF蛋白的表达。     

腺病毒介导VEGF基因在人骨髓基质干细胞中的表达

[目的] 体外培养人骨髓基质干细胞(hBMSc),将VEGF165基因腺病毒载体共转染hBMSc,观察其表达.[方法] 共转染实验分为3组:VEGF165和eGFP转染组、空腺病毒载体转染组和未转染组.取重组腺病毒Ad-VEGF165(MOI=20 pfu/cell)感染hBMSe.荧光显微镜下观察eGFP的表达,判断感染效率及细胞生长情况,RT-PCR检测,ELISA和Westernblot检测分别用于观察基因转染的表达.[结果] MOI=20 pfu的转染率已达80%以上,取此值为转染滴度.提取出的总RNA质量较好.以双链cDNA为模板扩增VEGF165基因,产物经1%琼脂糖凝胶电泳,均可见到549 bp(GAPDH)2条带.转染组可见VEGF165表达,空载体组和空白对照组未检测到VEGF165表达.ELISA结果显示基因转染组与转染空载体组和未转染组结果差异呈显著性(P<0.05).Western blot显示:转染组在50 KD处出现特征性条带,而空载体组和空白对照组未见条带.[结论] 外源性VEGF基因在人骨髓基质干细胞(hBMSc)内获得稳定表达,在本实验条件下培养的hBMSc具有典型成骨细胞的形态学特征和较强的生物学活性,向成骨细胞方向分化效率较高,细胞数量可以满足骨组织工程研究的要求.     

人BMP-7及VEGF基因共表达腺病毒的构建及在兔骨髓基质干细胞中的共表达

目的:构建人BMP-7及VEGF165基因共表达腺病毒并观察其在兔骨髓基质干细胞(rBMSC)中的表达.方法:利用AdEasy腺病毒表达系统构建人BMP-7及VEGF165基因共表达腺病毒AdB7V,体外感染rBMSC后通过观察绿色荧光蛋白表达及RT-PCR、免疫组化方法鉴定外源基因的表达.结果:rBMSC经AdB7V感染后72h可观察到绿色荧光蛋白(GFP)表达,RT-PCR、免疫细胞化学方法证实外源性BMP-7及VEGF165基因均可在rBMSC中表达.结论:成功构建人BMP-7及VEGF165基因共表达重组腺病毒,证实了其在rBMSC的表达,为骨再生的局部联合基因治疗打下基础.     

VEGF165和Ang-1协同改善急性心梗大鼠心功能

目的探讨血管内皮细胞生长因子和促血管生成素联合使用治疗大鼠急性心肌梗死及其机制.方法扩增携带人VEGF165或Ang-1的复制缺陷型病毒;制作大鼠急性心肌梗死模型,结扎后立即于缺血的心肌区域注射Ad-VEGF165或/和Ad-Ang-1,术后注射Brdu;免疫组织化学双染色观察新生内皮细胞和心肌细胞数;用超声心动图仪检测左心功能.结果 VEGF165和Ang-1联合使用可显著改善左心功能.梗死区发现新生心肌细胞,并且新生的内皮细胞和心肌细胞数显著多于其他组,梗死区细胞表达C-kit阳性.结论 VEGF165和Ang-1可显著改善急性心肌梗死大鼠心功能,其机制与骨髓造血干细胞的动员密切联系.     

血管内皮生长因子和促血管生成素及其受体Tie2在肝细胞癌发生发展中的作用

目的探讨血管内皮生长因子(VEGF)和促血管生成素(Angiopietin)及其受体Tie2在启动肝细胞癌(HCC)血管生成中的调控机制及在HCC发生发展中的作用。方法新鲜HCC标本及癌旁肝组织38例,用实时定量逆转录-聚合酶链反应(RT-PCR)的方法检测Ang-1、Ang-2、Tie2和VEGF在各组织标本中的表达,以CD34标记新生血管内皮并计数微血管密度(MVD),分析上述因子在HCC组织和非癌肝组织中的表达差异、相互作用及其与MVD、临床病理特征之间的关系。结果Ang-1、Tie2在HCC和非癌肝组织中的表达差异无统计学意义(0．194 7±0．086 2比0．232 6±0．109 8,1．601 6±0．900 7比1．340 0±0．703 7,P均>0．05),而VEGF和Ang-2在HCC组织的表达高于非癌肝组织(1．038 0±0．572 0比0.832 3±0．182 4,0．621 3±0.417 6比0．442 9±0．330 1,P均<0．05);VEGF、Ang-2、Ang-2／1都与MVD和临床病理特征相关(P<0．01),但与组织分化程度无关(P>0．05)。结论Ang-2／1的失衡表达及其与VEGF和Tie2的共同作用是启动肝组织血管生成并诱发HCC发生、发展的重要因素;这种因素在HCC中的持续作用进一步促进了肿瘤血管生成和恶性生物学行为。     

Tie2 ligands angiopoietin-1 and angiopoietin-2 are coexpressed with vascular endothelial cell growth factor in growing human bone

Angiogenesis is essential for bone growth and repair. Recent studies have shown that the endothelial-specific mitogen vascular endothelial growth factor (VEGF) is a key regulator of vascular invasion into the growth plate in infant and adolescent animals. In order to identify mechanisms regulating VEGF-induced angiogenesis in growing bone, we have investigated the expression of the angiopoietins (Ang-1 and Ang-2) in human neonatal ribs. Ang-1 and Ang-2 exhibited similar patterns of staining in the growing rib. In the cartilage, expression of Ang-1 and Ang-2 increased with chondrocyte maturation. Ang-1, Ang-2, and VEGF were not detected in the resting zone except adjacent to vascular canals, and maximum expression was detected at the cartilage bone interface. In the cartilage, Ang-2 was more highly expressed than Ang-1 or VEGF, with staining observed in the proliferating, hypertrophic, and mineralized zones. In the bone, Ang-1, Ang-2, and VEGF were detected in modeling and remodeling sites. Ang-1 was detected in the majority of osteoblasts, osteoclasts, and in some marrow space cells. Ang-2 was expressed at variable levels by osteoblasts and osteoclasts in modeling and remodeling bone. VEGF was detected in cells at bone surfaces and in the marrow spaces. Strong staining for VEGF was observed in osteoblasts and osteoclasts in modeling and remodeling bone. In the perichondrium, Ang-1, Ang-2, and VEGF were most highly expressed adjacent to the hypertrophic zone and at sites of bone collar formation. In the periosteum, Ang-1, Ang-2, and VEGF expression colocalized with alkaline phosphatase expression. These observations provide the first evidence for the expression of the angiopoietins in growing human bone in vivo. The distribution of Ang-1, Ang-2, and VEGF indicate these factors may play key roles in the regulation of angiogenesis at sites of endochondral ossification, intramembranous ossification, and bone turnover in the growing human skeleton.     

Expression of the Vascular Endothelial Growth Factor and Angiopoietins in Mucoepidermoid Carcinoma of Salivary Gland

Disrupted coordination of angiogenesis regulating signals, among them the vascular endothelial growth factor (VEGF) and angiopoietins (Angs), has been associated with abnormal angiogenesis and tumor progression. While VEGF induces endothelial cell proliferation, thereby initiating vessel formation, Angs are subsequently required for mural cell attachment, thus influencing remodeling and maturation of this vasculature. In addition to tumor cell, endothelial and mural cells, as well as myofibroblasts may also contribute to the secretion of these factors. In this study, we have analyzed by immunohistochemistry the expression of VEGF, Ang-1, Ang-2 and the Angs receptor Tie2 in both the stroma and tumor cells of mucoepidermoid carcinoma (MEC) of salivary gland. We have demonstrated that when myofibroblasts were detected adjacent to the cancer cells, they were frequently associated with intense positive staining for Ang-1 and Ang-2, and no reactivity to VEGF and Tie2. These myofibroblast-rich Ang-1 and Ang-2-stained areas were more commonly found in high-grade MEC cases than in low-grade ones. As for the malignant cells, they frequently expressed all proteins studied, but Ang-2 and VEGF were detected at higher levels compared to Ang-1 and Tie2. Our results indicate that the MEC environment favors cooperative activity between Angs and VEGF in modulating vascular growth and tumor aggressiveness.     

Angiopoietin-2, marker and mediator of endothelial activation with prognostic significance early after trauma?

OBJECTIVE: To measure plasma levels of angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2), and vascular endothelial growth factor (VEGF) early after trauma and to determine their clinical significance. BACKGROUND: Angiopoietins and VEGF play a central role in the physiology and pathophysiology of endothelial cells. Ang-2 has recently been shown to have pathogenetic significance in sepsis and acute lung injury. Little is known about the role of angiopoietins and VEGF early after trauma. METHODS: Blood specimens from consecutive major trauma patients were obtained immediately upon arrival in the emergency department and plasma samples assayed for Ang-1, Ang-2, VEGF, markers of endothelial activation, protein C pathway, fibrinolytic system, and complement. Base deficit was used as a measure of tissue hypoperfusion. Data were collected prospectively. RESULTS: Blood samples were obtained from 208 adult trauma patients within 30 minutes after injury before any significant fluid resuscitation. Plasma levels of Ang-2, but not Ang-1 and VEGF were increased and correlated independently with severity of injury and tissue hypoperfusion. Furthermore, plasma levels of Ang-2 correlated with markers of endothelial activation, coagulation abnormalities, and activation of the complement cascade and were associated with worse clinical outcome. CONCLUSIONS: Ang-2 is released early after trauma with the degree proportional to both injury severity and systemic hypoperfusion. High levels of Ang-2 were associated with an activated endothelium, coagulation abnormalities, complement activation, and worse clinical outcome. These data indicate that Ang-2 is a marker and possibly a direct mediator of endothelial activation and dysfunction after severe trauma.     

Angiogenesis in the brain during development: the effects of vascular endothelial growth factor and angiopoietin-2 in an animal model

OBJECT: The goal of this study was to examine the roles of vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) in the formation of blood vessels in the brain in a developmental animal model not routinely used for such a study. METHODS: Either VEGF, Ang-2, or a combination of the two factors were injected into the optic tectum of 4-day-old quail embryos. Immunohistochemical analysis and laser confocal microscopy were used to observe the effects on endothelial cells in the brain. Vascular endothelial growth factor and Ang-2 had very different effects on the development of blood vessels; the former caused expansion and the latter retraction of these vessels. Treatment with a combination of VEGF and Ang-2 caused retroorbital or intraventricular hemorrhage, and brain blood vessels appeared enlarged and dysmorphic, with dramatically extended filopodia. CONCLUSIONS: Some of these observations may provide insight into how one may develop a better model of brain arteriovenous malformations.     

Ang-1基因转染的内皮祖细胞移植对大鼠肾脏缺血再灌注后的保护作用

目的 评估Ang-1基因转染的内源性内皮祖细胞(endothelial progenitor cells,EPCs)移植治疗肾脏缺血再灌注损伤的作用,进一步完善EPCs对缺血肾脏产生保护作用的旁分泌机制.方法利用重组腺病毒载体Ad-Ang-1感染EPCs,并进行转染效率鉴定,进一步移植治疗大鼠缺血肾脏,术后72 h给予肾功能评估,并检测大鼠缺血肾脏中VEGF、Ang-1以及Ang-2表达情况.结果EPCs移植治疗后,大鼠肾脏功能明显改善,术后72 h检测大鼠患肾,其内VEGF、Ang-1以及Ang-2表达均明显提升.结论 Ang-1基因转染的EPCs移植治疗可以有效地治疗大鼠肾脏IRI,其作用机制可能与EPC归巢后旁分泌过量Ang-1相关,并且促使肾脏血管新生.     

血管新生调控因子及其受体随增龄在小鼠肾组织内的表达

目的 探讨血管内皮生长因子（VEGF）及其受体（VEGFR）、血管生成素（Ang-1、Ang-2）及其受体（Tie2）在增龄小鼠肾脏中的表达,以及它们在肾脏衰老中的作用。 方法 选取4月龄、9月龄、12月龄及20月龄C57小鼠各6只,留取尿液及血液标本,应用常规生化法测定各组小鼠肾功能。用PAS染色对各组小鼠进行肾脏病理染色及分析;荧光染料肾脏灌注对肾小球毛细血管丛密度进行分析。应用免疫组化、免疫荧光、Western印迹及实时定量PCR方法分析肾脏转化生长因子（TGF-β1）及VEGF、VEGFR2（血管内皮生长因子受体2,Flk-1）、Ang-1、Ang-2、Tie2的蛋白及基因表达的变化。 结果 随着增龄,小鼠肾小球硬化指数（GSI）增加,其中20月龄约为4月龄的5倍（P < 0.05）。荧光染料肾脏灌注后见肾小球毛细血管内荧光强度呈减少趋势,20月龄显著低于4月龄（P < 0.05）。免疫组化结果显示,TGF-β1在肾小球及肾小管间质中的表达呈增多趋势,且以肾小球内增多较显著,各组间差异有统计学意义（P < 0.05）。免疫荧光结果显示,Ang-1在肾小球内的表达呈减少趋势,20月龄较4月龄显著减少（P < 0.05）。实时定量PCR结果显示,不同月龄小鼠肾组织内VEGF、Flk-1、Ang-1、Ang-2、Tie2的mRNA表达水平均呈下降趋势,20月龄与4月龄间VEGF、Flk-1及Ang-2差异无统计学意义,而Ang-1和Tie2差异有统计学意义（均P < 0.05）。Western印迹结果显示,4组小鼠肾脏VEGF、Flk-1、Ang-1、Ang-2、Tie2的蛋白表达水平也呈下降趋势,且20月龄与4月龄差异均有统计学意义（均P < 0.05）;TGF-β1的表达量呈增多趋势,其中20月龄较4月龄增多约40%（P < 0.05）。相关分析结果显示,肾小球毛细血管密度、Ang-1、Ang-2、Tie2、VEGF、Flk-1的mRNA及蛋白水平均与Scr呈负相关。 结论 随衰老程度加重,小鼠肾组织内血管内皮生长因子及其受体、血管生成素及其受体的表达减少,且与增龄肾脏的形态学改变及肾功能改变相关。