仙人掌提取物对浅Ⅱ度烫伤治疗作用的研究

目的:探讨野生仙人掌提取物对浅Ⅱ度烫伤小鼠的治疗作用。方法:建立浅Ⅱ度烫伤小鼠模型,随机分为仙人掌提取物乙醇组A1,蒸馏水组A2和混合组A3(每组又分低、中、高3个浓度),烫伤(阴性)组B,药物(阳性)组C。每天涂药1次,观察、记录创面愈合情况,分别在5d和12d各组随机取5只小鼠做病理切片,观察并记录血管、纤维细胞、成纤维细胞等的数量,进行统计学分析。结果:①混合提取物中、高浓度组愈合时间,与烫伤组相比有显著性差异(p0.05、p0.01)。②5d时不同的仙人掌提取物组与烫伤组比,血管数量、成纤维细胞占总纤维细胞的百分含量都有差异(p0.05)。结论:野生仙人掌提取物能加速烫伤创面愈合,尤其对血管和成纤维细胞有显著的促进增生作用,并加速再上皮化。     

外源性VEGF促进大鼠Ⅱ度烫伤创面中晚期愈合

目的探究Ⅱ度烫伤时外源性血管内皮生长因子(vascular endothelial growth factor,VEGF)对皮肤愈合及其对表皮干细胞(epidermal stem cells,ESCs)迁移、分化的影响方法健康Wistar大鼠随机分为VEGF组、空白对照组、阿西替尼(Axitinib,VEGF抑制剂)组。采用水浴烫伤法制备Ⅱ度烫伤模型,分别以0.2μg/ml VEGF、PBS溶液和10μg/ml阿西替尼处理各组创面,各组均治疗7d,从烫伤至创面愈合分别在第2d、8d、14d及21d测量创面愈合情况,并取创面组织作组织学检测,运用免疫组化技术检测ECSs的分布及数量。结果①创面愈合率:烫伤后21d VEGF组>对照组>阿西替尼组;②愈合速度:烫伤后1-7d空白对照组>阿西替尼组>VEGF组,其后VEGF组愈合速度逐渐加快,第14d开始愈合速度表现为VEGF组>空白对照组>阿西替尼组;③组织学变化:烫伤后8-21d,VEGF组表皮细胞增殖明显,表皮修复和毛囊再生迅速,均早于空白对照组及阿西替尼组。④ECSs阳性细胞率变化:烫伤后第8-14dVEGF组ECSs阳性细胞率明显高于空白对照组和阿西替尼。结论Ⅱ度烫伤时,外源性VEGF在愈合中晚期加快愈合速度使愈合时间明显缩短,并且促进毛囊汗腺的再生使修复后的创面在外观、功能与正常皮肤相近,有助于提高全层皮肤创面的愈合质量。     

慢性复合应激对成年海马FGF-2的表达及神经发生的影响

目的通过观察慢性复合应激后大鼠海马FGF-2表达的变化,来探讨慢性复合应激对FGF-2表达的影响及其与海马神经发生的联系。方法成年雄性大鼠随机分为复合应激组和正常对照组。复合应激组动物每天交替无规律暴露于复合应激原中达6周。然后运用免疫组织化学方法、Western-blot和RT-PCR技术观察海马FGF-2表达的变化。结果慢性复合应激组动物海马FGF-2阳性细胞的表达量增多(P<0.05);海马FGF-2蛋白的表达明显增加(P<0.05);海马FGF-2 mRNA水平明显上调(P<0.05)。结论慢性复合应激可引起海马FGF-2阳性细胞表达量增加和FGF-2表达水平升高,提示应激后内源性FGF-2的表达增高可能是慢性复合应激促海马神经发生的因素之一。     

湿润烧伤膏联合重组人碱性成纤维细胞生长因子对浅Ⅱ度烧伤患者创面肉芽组织HIF-1α、VEGF蛋白表达的影响

摘要 目的：观察湿润烧伤膏联合重组人碱性成纤维细胞生长因子（rh-bFGF）凝胶治疗浅Ⅱ度烧伤患者的疗效及对创面肉芽组织缺氧诱导因子-1α（HIF-1α）、血管内皮生长因子（VEGF）蛋白表达的影响。方法：选取天津市第一中心医院2018年12月到2020年12月之间收治的浅Ⅱ度烧伤患者70例,根据住院号单双数分为对照组和实验组两组,各35例。对照组给予rh-bFGF凝胶治疗,实验组给予湿润烧伤膏联合rh-bFGF凝胶治疗,两组均治疗2周。对比两组疗效、创面愈合率、症状消失时间、不良反应发生率、血清炎症因子水平变化和创面肉芽组织HIF-1α、VEGF蛋白表达变化。结果：实验组的临床总有效率、创面愈合率均高于对照组（P<0.05）。实验组的肿胀、疼痛、红斑、水疱、渗液症状消失时间均短于对照组（P<0.05）。治疗2周后,两组患者血清白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、超敏C反应蛋白（hs-CRP）水平较治疗前下降（P<0.05）,且实验组低于对照组（P<0.05）。治疗2周后,实验组患者创面肉芽组织VEGF蛋白表达水平高于对照组,HIF-1α蛋白表达水平低于对照组（P<0.05）。对照组的不良反应发生率为11.43%,与实验组的5.71%对比差异无统计学意义（P>0.05）。结论：与单用rh-bFGF凝胶治疗相比,联合湿润烧伤膏治疗浅Ⅱ度烧伤患者可更好地促进创面愈合,改善患者症状,同时还可改善血清炎症因子水平及创面肉芽组织HIF-1α、VEGF蛋白表达,安全可靠。     

在烫伤条件下VEGF对大鼠皮肤eNOS基因表达的影响

目的:探讨大鼠烫伤愈合过程中VEGF对皮肤组织eNOS基因表达的影响。方法:采用了大鼠深II度烫伤模型,分别以VEGF和阿西替尼(VEGF抑制剂)干预,观察大鼠烫伤后愈合过程的组织学变化,PCR检测创面eNOS基因的表达。结果:肉眼观察VEGF组大鼠创面最早愈合,对照组大鼠愈合速度其次,阿西替尼组创面愈合最晚;HE染色结果显示伤后第2、8、21天VEGF组炎性细胞浸润程度、新生毛细血管数量均高于对照组和阿西替尼组,PCR结果显示烫伤后第2、8天VEGF组的eNOS基因的表达明显上调,且同时间点与对照组比较有统计学意义(P0.05)。结论:VEGF可诱导烫伤创面eNOS基因的表达,增加血管生成重要因子NO的生成,有利于创面愈合。     

基于光学相干层析成像技术的鼠皮肤II度烫伤及其修复过程监控

建立小鼠皮肤的Ⅱ度烫伤的模型,分别于烫伤前、烫伤后1 min、8 h、1 d、3 d、5 d、7 d、9 d、11 d、13 d、15 d、17 d,用光学相干层析成像技术(OCT)对烫伤的创面进行动态的监测,同时对创面进行组织病理学的检测,对结果进行分析。利用OCT单次散射模型获得皮肤在烫伤前后光衰减系数的变化情况,结果显示光衰减系数的值在烫伤后是呈先减小后增大的趋势,待修复完全后又恢复到烫伤前相近的值。该结果与组织病理学的结果相吻合,这表明OCT技术可以用于实时诊断烫伤的皮肤。     

獾油促进深Ⅱ度烫伤小鼠创面愈合作用的研究

目的:检测獾油对小鼠深Ⅱ度烫伤创面愈合的促进作用.方法:制备小鼠深Ⅱ度烫伤模型,分为烫伤对照组(A组)、植物油治疗组(B组)和獾油治疗组(C组).创面依次用生理盐水纱布、植物油纱布、獾油纱布覆盖.无菌纱布包扎固定,每日换药一次.于伤后7天、10天、15天按照Nagelschmidt法观察并记录创面愈合面积,计算创面愈合率;取创面组织,制成石蜡切片,观察病理及组织形态学变化.结果:獾油能加速烫伤创面的再上皮化,促进创面的愈合;提高烫伤组织细胞的增殖活性;促进烫伤创面表皮干细胞的增殖分化.结论:獾油对深Ⅱ度烫伤创面愈合有促进作用     

麦冬不同提取物对过氧化氢损伤人血管内皮细胞ICAM-1、VEGF、Bcl-2表达的影响

目的:观察麦冬不同提取物对过氧化氢诱导的人脐静脉内皮细胞(HUVEC)间黏附分子-1(ICAM-1)和VEGF、Bc(?)-2表达的影响。方法:体外培养HUVEC,用过氧化氢(H_2O_2)制造HUVEC损伤模型。以四甲基偶氮唑蓝(MTT)比色法检测细胞存活数量,用流式细胞仪检测HUVEC表面ICAM-1的表达量;免疫细胞化学方法检测HUVEC的VEGF、Bc(?)-2的分布情况。结果:模型组较正常对照组细胞增殖活性明显降低(P<0.01)。与模型组相比,经麦冬水提物、正丁醇提取物处理组细胞增殖活性明显增加(P<0.05,P<0.01)。流式细胞仪检测显示正丁醇提取物可降低过氧化氢增加的ICAM-1基因的表达。Bc(?)-2的表达,模型组明显低于正常对照组,而正丁醇组表达明显高于模型组(P<0.01)。VEGF的表达,模型组明显高于正常对照组,麦冬水提物、正丁醇提取物处理组高于模型组(P<0.05,P<0.01)。结论:麦冬提取物具有抗凋亡、促增殖、降低细胞间黏附分子-1表达的作用,尤以正丁醇提取物效果更为显著。     

异种脱细胞真皮基质联合丝素蛋白辅料对幼儿Ⅱ度烫伤的临床疗效研究

目的分析异种脱细胞真皮基质联合丝素蛋白辅料对幼儿Ⅱ度烫伤的治疗效果。 方法随机选择湖北医药学院附属襄阳市第一人民医院104例Ⅱ度烫伤患儿,采用随机数字表法分为对照组（丝蛋白敷料覆盖）和试验组（异种脱细胞真皮基质联合丝蛋白敷料覆盖）,每组52例。实验结果中计量资料用 ±s表示,组间比较采用两样本t检验,治疗前后比较采用配对t检验;计量资料采用百分比表示,比较采用χ²检验,疼痛程度采用Z检验。 结果试验组患儿换药次数（2.87±0.98）?次、完全溶痂时间（8.26±3.18）d、温哥华评分（4.25±0.87）分,对照组分别为（5.64±1.02）次、（12.38± 3.02）d、（8.12±1.04）分,差异均有统计学意义（t = 14.121,6.775,20.582,P均< 0.001）。试验组患儿浅Ⅱ度创面感染率0.00%、愈合时间（8.53±1.49）d,对照组患儿分别为17.31%、（10.61±2.05） d,差异均有统计学意义（χ² = 9.853,P = 0.002,t = 5.918,P < 0.001）。试验组患儿深Ⅱ度创面感染率5.77%、愈合时间（12.90±1.64）d,对照组患儿分别为34.62%、（15.45±1.49）d,差异均有统计学意义（χ² = 13.425,P < 0.001,t = 8.299,P < 0.001）。治疗后比较,试验组患儿创面周围炎症评分（1.91±0.48）分、创面渗出物评分（2.86±0.45）?分,对照组患儿分别为（1.33±0.37）分、（2.09±0.53）分,差异有统计学意义（t = 6.901,7.986,P均?< 0.001）。试验组患儿疼痛程度与对照组比较,差异有统计学意义（Z = 41.657,P < 0.001）。 结论采用异种脱细胞真皮基质联合丝蛋白敷料覆盖治疗Ⅱ度烫伤患儿,可减少术后感染,缩短痊愈时间,降低疼痛程度,疗效优势明显。     

麦冬不同提取物对过氧化氢损伤人血管内皮细胞ICAM-1、VEGF、Bcl-2表达的影响

目的：观察麦冬不同提取物对过氧化氢诱导的人脐静脉内皮细胞（HUVEC）间黏附分子-1（ICAM-1）和VEGF、Bcl-2表达的影响。方法：体外培养HUVEC,用过氧化氢（H2O2）制造HUVEC损伤模型。以四甲基偶氮唑蓝（MTT）比色法检测细胞存活数量,用流式细胞仪检测HUVEC表面ICAM-1的表达量;免疫细胞化学方法检测HUVEC的VEGF、Bcl-2的分布情况。结果：模型组较正常对照组细胞增殖活性明显降低（P〈0．01）。与模型组相比,经麦冬水提物、正丁醇提取物处理组细胞增殖活性明显增加（P〈0．05,P〈0．01）。流式细胞仪检测显示正丁醇提取物可降低过氧化氢增加的ICAM-1基因的表达。Bcl-2的表达,模型组明显低于正常对照组,而正丁醇组表达明显高于模型组（P〈0．01）。VEGF的表达,模型组明显高于正常对照组,麦冬水提物、正丁醇提取物处理组高于模型纽（P〈0．05,P〈0．01）。结论：麦冬提取物具有抗凋亡、促增殖、降低细胞间黏附分子-1表达的作用,尤以正丁醇提取物效果更为显著。     

AMP-activated protein kinase positively regulates FGF-2-stimulated VEGF synthesis in osteoblasts

AMP-activated protein kinase (AMPK) is recognized as a regulator of energy homeostasis. We have previously reported that basic fibroblast growth factor (FGF-2) stimulates vascular endothelial growth factor (VEGF) release through the activation of p44/p42 mitogen-activated protein (MAP) kinase and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) in osteoblast-like MC3T3-E1 cells. In the present study, we investigated the involvement of AMPK in FGF-2-stimulated VEGF release in these cells. FGF-2 time-dependently induced the phosphorylation of AMPK α-subunit (Thr-172). Compound C, an AMPK inhibitor, which suppressed the FGF-2-induced phosphorylation of AMPK, significantly inhibited the VEGF release stimulated by FGF-2. The AMPK inhibitor also reduced the mRNA expression of VEGF induced by FGF-2. The FGF-2-induced phosphorylation of both p44/p42 MAP kinase and SAPK/JNK was attenuated by compound C. These results strongly suggest that AMPK positively regulates the FGF-2-stimulated VEGF synthesis via p44/p42 MAP kinase and SAPK/JNK in osteoblasts.     

VEGF is a chemoattractant for FGF-2-stimulated neural progenitors

Migration of undifferentiated neural progenitors is critical for the development and repair of the nervous system. However, the mechanisms and factors that regulate migration are not well understood. Here, we show that vascular endothelial growth factor (VEGF)-A, a major angiogenic factor, guides the directed migration of neural progenitors that do not display antigenic markers for neuron- or glia-restricted precursor cells. We demonstrate that progenitor cells express both VEGF receptor (VEGFR) 1 and VEGFR2, but signaling through VEGFR2 specifically mediates the chemotactic effect of VEGF. The expression of VEGFRs and the chemotaxis of progenitors in response to VEGF require the presence of fibroblast growth factor 2. These results demonstrate that VEGF is an attractive guidance cue for the migration of undifferentiated neural progenitors and offer a mechanistic link between neurogenesis and angiogenesis in the nervous system.     

Effect of FGF-1 and FGF-2 on VEGF binding to human umbilical vein endothelial cells

When FGF-1 or FGF-2 and VEGF were added together, the mitogenic effect of FGF-1 or FGF-2 and VEGF on HUVEC was additive. However, when HUVECs were preincubated for 2 days with 10 ng/ml FGF-1 in the absence of VEGF, the Scatchard plot of [125I]VEGF binding sites was shifted to the right: both affinity classes of VEGF binding sites were equally affected, such that the total number of sites increased twofold. It is suggested that this type of interaction may be related to tumor angiogenesis and wound repair.     

Up-regulation by zinc of FGF-2-induced VEGF release through enhancing p44/p42 MAP kinase activation in osteoblasts

We previously reported that basic fibroblast growth factor (FGF-2) activates stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) and p44/p42 mitogen-activated protein (MAP) kinase resulting in the stimulation of vascular endothelial growth factor (VEGF) release in osteoblast-like MC3T3-E1 cells. In the present study, we investigated whether zinc affects the VEGF release by FGF-2 in MC3T3-E1 cells. The FGF-2-induced VEGF release was significantly enhanced by ZnSO(4) but not Na(2)SO(4). The enhancing effect of ZnSO(4) was dose-dependent between 1 and 100 muM. ZnSO(4) markedly enhanced the FGF-2-induced phosphorylation of p44/p42 MAP kinase while having little effect on the SAPK/JNK phosphorylation. PD98059 significantly reduced the amplification by ZnSO(4) of the FGF-2-stimulated VEGF release. Taken together, our findings strongly suggest that zinc enhances FGF-2-stimulated VEGF release resulting from up-regulating activation of p44/p42 MAP kinase in osteoblasts.     

肝素促进深Ⅱ度烧伤小鼠bFGF及PDGF表达的研究

目的:探讨肝素对深Ⅱ度烧伤小鼠内源性碱性成纤维生长因子(bFGF)和血小板源生长因子(PDGF)表达的影响.方法:制作小鼠深Ⅱ度烫伤模型,肝素组用3000U/mL肝素外敷伤口2min,每日2次,观察创面愈合状况,并在给药后第3天分别从肝素组(烫伤后用肝素处理)、烫伤组(烫伤后自然愈合)及对照组(未进行烫伤处理)取5只小鼠处死,提取烫伤及其周围皮肤组织的蛋白质,用Western-blotting检测bFGF和PDGF的表达状况.结果:肝素组bFGF和PDGF表达均高于烫伤组;而烫伤组的两种生长因子的表达均高于对照组.结论:烫伤后bFGF和PDGF的表达量增高,且肝素能促进深Ⅱ度烧伤内源性生长因子bFGF、PDGF的表达.     

The distribution of the growth factors FGF-2 and VEGF, and their receptors, in growing red deer antler

The cellular distributions of the growth factors FGF-2 and VEGF, and their receptors FGFR1, FGFR2 and FGFR3, and VEGFR-2 respectively, were visualized by immunohistochemistry and light microscopy in sections of growing red deer antler. Both of these signalling systems were widely expressed in the integument and osteocartilaginous compartments. FGF-2 was found in the same cells as all three FGFRs, indicating that FGF signalling may be principally autocrine. The patterns of labelling for VEGF and its receptor were similar to those seen for FGF-2 and FGFR-3, in both compartments. Our data are consistent with the findings of others in suggesting that FGF-2 induces expression of VEGF, to stimulate and maintain high rates of neovascularisation and angiogenesis, thereby providing nutrients to both velvet and bone as they rapidly grow and develop. The presence of FGF and VEGF and their receptors in epithelial cells suggests that these signalling systems play a role in skin development, raising the possibility that one or both may be involved in the close coupling of the coordinated growth of the integument and osteocartilage of antler, a process which is poorly understood at present.     

Ephrin-B2促进大鼠局灶脑缺血再灌注后VEGF表达及血管新生

目的:探讨Ephrin-B2对大鼠脑缺血再灌注后脑组织中血管新生的调节作用及其可能的机制。方法:雄性SD大鼠随机分为正常组及、缺血再灌注组及Ephrin-B2干预组,后两组再分为4天、7天、14天、28天亚组;线栓法制备局灶性大脑中动脉缺血再灌注模型;改良神经功能评分(modified neurological severity scores mNSS)评分法对各时间点模型进行评分;Western blot及荧光定量PCR检测缺血脑组织中血管内皮生长因子(Vascular Endothelial Growth Factor VEGF)的表达;以免疫荧光双标法定位VEGF表达的细胞类型;以CD31+BrdU计数缺血半暗带中新生微血管密度(microvessel densityMVD)。结果:Ephrin-B2干预组与缺血再灌注组各时间点亚组比较,新生微血管密度测定计数较缺血再灌注组均显著增加(P0.05),神经功能评分均显著降低(P0.05),VEGF mRNA水平及蛋白表达水平均显著增加(P0.05),VEGF主要表达于CD31阳性的血管内皮细胞。结论:Ephrin-B2通过上调VEGF的表达促进脑缺血再灌注后缺血半暗带血管新生,从而促进神经功能缺失的修复。     

ELV对青春期大鼠睾丸中VEGF与VEGFR2蛋白表达的影响

目的研究血管内皮生长因子(VEGF)及其受体2(VEGFR2)在实验性左侧精索静脉曲张大鼠睾丸中的表达和定位,探讨精索静脉曲张中VEGF和VEGFR2的可能作用。方法通过部分结扎左肾静脉建立大鼠实验性左侧精索静脉曲张模型,于术后2周和4周取材,采用免疫组化法检测VEGF、VEGFR2在睾丸上的表达变化。结果 ELV2周与4周组大鼠两侧睾丸中VEGF蛋白表达均上调,但ELV组间VEGF蛋白表达没有明显变化;ELV2周组大鼠睾丸中VEGFR2蛋白的表达与对照组比较增强,而4周组比对照组和2周组均显著增强。结论实验性左侧精索静脉曲张对VEGF、VEGFR2蛋白的表达有影响,说明它们与男性不育可能有一定的关系。     

Neurons produce FGF2 and VEGF and secrete them at least in part by shedding extracellular vesicles

We previously found that neurons are able to affect the ability of brain capillary endothelial cells to form in vitro a monolayer with properties resembling the blood-brain barrier. We then looked, by immunofluorescence and western analysis, for factors, produced by neurons, with the potential to influence growth and differentiation of endothelial cells. In the present paper, we report that neurons produce both vascular endothelial growth factor and fibroblast growth factor 2, two well-known angiogenic factors. More interestingly, we gained evidence that both factors are released by neurons, at least in part, by shedding of extracellular vesicles, that contain beta1 integrin, a membrane protein already known to be part of extracellular vesicles released by tumour cells. Shedding of extracellular vesicles by neurons was also confirmed by scanner electron microscopy.