Effect of Recombinant Human Epidermal Growth Factor Against Cutaneous Scar Formation in Murine Full-thickness Wound Healing

A visible cutaneous scar develops from the excess formation of immature collagen in response to an inflammatory reaction. This study examined the role of epidermal growth factor (EGF) in the formation of cutaneous scars. Twenty Crl:CD-1 (ICR) mice were used and 2 full-thickness skin wounds were made on the dorsum of each mouse. One of the wounds was treated with recombinant human EGF by local application and the other was treated with saline for control until complete healing was achieved. The EGF-treated group''s wounds healed faster than the control group''s. The width of the scar was smaller by 30% and the area was smaller by 26% in the EGF-treated group. Inflammatory cell numbers were significantly lower in the EGF-treated group. The expression of transforming growth factor (TGF)-β₁ in the EGF-treated group was increased. It was observed that the amount of collagen in the EGF-treated group was larger than the control group. In the EGF-treated group, the visible external scars were less noticeable than that in the control group. These results suggest that EGF can reduce cutaneous scars by suppressing inflammatory reactions, decreasing expression of TGF-β₁, and mediating the formation of collagen.     

Cyclical Cell Stretching of Skin-Derived Fibroblasts Downregulates Connective Tissue Growth Factor (CTGF) Production

Delayed healing of skin wounds can be caused by wound instability, whereas appropriate massage or exercise prevents sclerosis and scar contracture. However, the mechanism by which wound healing is related to mechanical stress has not been fully elucidated. The present study aimed to identify whether mechanical stretching of fibroblasts reduces their production of extracellular matrix. We transferred skin fibroblasts into collagen-coated elastic silicone chambers, cultured them on a stretching apparatus, and used RT-PCR to examine the effects of mechanical stretching on the expression levels of 17 genes related to extracellular matrix production and growth factor secretion. We found that connective tissue growth factor (CTGF) was downregulated after 24 hr of cell stretching. Specifically, the CTGF mRNA and protein levels were 50% and 48% of the control levels, respectively. These findings suggest that cyclic stretching of fibroblasts contributes to anti-fibrotic processes by reducing CTGF production.     

结缔组织生长因子与肺纤维化

最近,结缔组织生长因子(CTGF)以其在促纤维化中的决定性作用而受到广泛关注。它由TGF-β介诱导产生,是其下游效应介质,主要介导TGF-β的促细胞增生和细胞外基质合成,生物学效应较单一。肺纤维化是多种原因引起慢性肺疾病的共同结局,随着研究的深入,人们对CTGF促肺纤维化作用机制的认识越来越深刻,近年来,针对拮抗CTGF以治疗肺纤维化已做了大量研究,CTGF可能成为治疗纤维化疾病的新的分子靶点。     

结缔组织生长因子在大鼠肺纤维化模型中的蛋白表达

目的 :了解结缔组织生长因子 (connectivetissuegrowthfactor ,CT GF)在大鼠肺纤维化模型中的表达状况及其与肺纤维化之间的关系。方法 :42只Wistar大鼠随机分为二组。模型组 (M组 ) :气管内灌注博莱霉素 ,对照组 (C组 ) :气管内灌注生理盐水。于灌注后第 7、14、2 8天每组分别处死大鼠 7只。制备大鼠肺组织切片。用免疫组化SABC法分别检测二组大鼠肺组织中CTGF及转化生长因子 β1 (TGF β1 )、纤维连接蛋白 (FN )在肺内的表达水平。结果 :M组CTGF及TGF β1 、FN在肺内的表达水平在各时间点均显著高于C组 (P <0 .0 5 )。结论 :CTGF作为TGFβ1 的下游因子 ,可能通过促进细胞外基质(ECM)如FN和胶原蛋白等合成而在博莱霉素诱导的肺纤维化形成过程中起重要作用     

Growth Factors in Intervertebral Disc Homeostasis

Intervertebral disc degeneration is a complex age-related pathology associated with back pain. Research on the growth factors that regulate disc homeostasis is of critical importance for understanding the basis of the disease. Here we summarize the data on the expression and function of various growth factors in the disc from in vivo and in vitro studies, as well as on their alterations during degeneration and ageing. Such studies are becoming more crucial in the prospect of clinical application of growth factors for the treatment of disc degeneration.     

胸腹水组织因子及组织因子途径抑制物的检测及其意义

目的研究三组疾病胸腹水组织因子(Tissue factor,TF)及组织因子途径抑制物(Tissue factor pathway inhibitor,TFPI)的表达及其鉴别诊断意义。方法TF和TFPI采用ELISA法测定抗原表达。结果胸腹水TF水平和TFPI水平,恶性肿瘤组(570.04±627.53)ng/L,(28.60±15.57)μg/L和结核病组(283.82±143.16)ng/L,(31.16±12.26)μg/L明显高于肝硬化组(60.83±66.87)ng/L,(7.84±5.45)μg/L,P<0.01。TF/TFPI比值则为恶性肿瘤组(32.17±44.19)明显高于结核病组(13.55±13.15)和肝硬化组(11.22±9.05,P<0.05)。在恶性肿瘤组中,胸腹水癌细胞阳性组的TF表达(1106.92±1244.28)ng/L高于阴性组(331.08±295.84)ng/L,P<0.05。而阳性组的TFPI水平(27.35±17.75)μg/L与阴性组(30.34±13.20)μg/L无明显差异(P>0.05)。TF/TFPI比值则为阳性组(59.59±65.10)明显高于阴性组(11.54±8.37,P<0.01)。结论检测胸腹水TF和TFPI并分析TF/TFPI比值可以作为临床实验室有鉴别诊断意义的辅助指标,同时还可了解疾病的某些病理机制,尤其是肿瘤的某些生物学行为。     

Platelet-Derived Growth Factor B-Chain of Hematopoietic Origin Is Not Necessary for Granulation Tissue Formation and Its Absence Enhances Vascularization

The hypothesis that wound repair is augmented by delivery of platelet-derived growth factor (PDGF) from platelets and macrophages is an attractive extrapolation from the known activities of PDGF in cell culture and in vivo. To test this hypothesis in mice, we prepared hematopoietic chimeras, in which the hematopoietic system of a normal adult mouse was replaced by the hematopoietic system of a PDGF B-chain -/- or +/+ donor. We initiated local granulation tissue formation either by implanting small surgical sponges to elicit a foreign body granulation tissue response, or by ligating the left common carotid to form an organized thrombus. We found that the absence of hematopoietic PDGF B-chain did not decrease the extent of granulation tissue or vascular lesion formation, and that the vascularization of both lesions increased by approximately 100%. We conclude that PDGF B-chain from cells of hematopoietic origin, including platelets and macrophages, is not important for granulation tissue formation, and that it reduces vascularization of granulation issue, probably through disabling of the short-range chemotactic gradients of PDGF that are important for recruiting pericytes/smooth muscle cells to the endothelium of new vessels.     

血管组织工程相关生长因子的控制释放研究进展

生长因子在细胞的黏附、增殖和组织的再生过程中发挥着重要的作用。将生长因子负载于高分子材料支架上，可以实现对生长因子的控制释放。其释放机制随负载方法的不同而存在着差异。本文总结了共混、凝胶、微球包埋以及化学键合法在血管组织工程中相关生长因子控制释放方面的研究进展，并指出了超细纤维包埋法的应用前景。     

Dose Responsive Effects of PDGF-BB,PDGF-AA,EGF, and bFGF on Granulation Tissue in a Guinea Pig Partial Thickness Skin Excision Model

A guinea pig partial thickness skin excistoti model wx used to evaluatc the effects of recombinant human PDGF-BB, 1′17GF-AA, ECF, and bFGF on granuLition tissue (neodermis) formation. These growth factors tended to increcisr the thickness of the granulation tissue bed when assesstd histcilogitally at day 7. Using only four animals per group, PDGF-BB at 30 and 100μg/ml consistently and significantly incwaseil the thickness of the granulation bed 2–3 limits that of control. Except for the increased thickness, the granulation tissue appmred normal. I'DGF-AA and EGF also significantly increased the granulation tissue thickness, and bFGF gave indications of an effect. Therc was no evidence of synergistic effects bthwxi PDGF-BB, EGF, and/or bFCF.     

当归注射液及前列腺素E_2对结缔组织生长因子在大鼠肺纤维化模型中表达作用的比较研究

目的探讨结缔组织生长因子(CTGF)在大鼠肺间质纤维化中的表达及其在25%当归注射液、前列腺素E2(dmPGE2)的肺脏保护作用中的意义。方法SD大鼠随机分为对照组、模型组、当归组、PGE2组。按博莱霉素(BLM)5mg/kg体重复制大鼠肺纤维化模型,当归组、dmPGE2组术后分别给于25%当归注射液(腹腔内注射)、dmPGE2(肌肉注射),于第28天处死全部大鼠。制备大鼠肺组织切片,用原位杂交及免疫组化SABC法分别检测CTGF mRNA、纤维连结蛋白(FN)和Ⅲ型胶原(Col-Ⅲ)在肺内的表达,HE染色评价肺纤维化程度。结果模型组CTGF mRNA、FN和Col-Ⅲ的表达及肺纤维化程度明显高于对照组(P<0.01),而当归组及PGE2组明显低于模型组(P<0.01)。两治疗组间比较,除CTGF外,其余指标无显著性差异(P>0.05),各组指标的相关分析表明,CTGF表达水平与肺纤维化程度(r=0.886;P<0.01)、FN(r=0.836;P<0.01)和Col-Ⅲ(r=0.918;P<0.01)呈正相关关系。结论25%当归注射液可能通过下调CTGF而减轻肺间质纤维化。     

高氨基酸血症对肾小球系膜细胞结缔组织生长因子表达的影响及与氧化应激的关系

目的研究高氨基酸血症对肾小球系膜细胞结缔组织生长因子(connective tissue growth factor,CTGF)表达的影响,探讨氧化应激在其中的作用。方法大鼠肾小球系膜细胞分为不同培养环境下的对照组(C组)、高氨基酸组(HA组)、维生素E组(HAE组)。培养24h后用荧光染料2',7'-Dichloroluores-cin diacetate(20μmol/L)测定各组细胞内活性氧产物(reactive oxygen specie,ROS)的生成情况从而反应细胞内氧化应激水平,培养24h后采用RT-PCR检测各组CTGF mRNA表达,培养48h后采用Western印迹检测各组CTGF蛋白的表达。结果HA组氧化应激水平、CTGF mRNA及蛋白的表达均明显高于C组(P<0.05);HAE组其氧化应激水平、CTGF mRNA及蛋白表达较HA组明显降低(P<0.05),且与C组比较差异无统计学意义(P>0.05)。结论高氨基酸血症可使肾小球系膜细胞氧化应激水平升高并产生过多的CTGF,抗氧化剂维生素E可减轻系膜细胞氧化应激及CTGF的表达,提示高氨基酸诱导的CTGF表达增多可能与氧化应激有关。     

结缔组织生长因子在血吸虫病肝纤维化鼠肝窦内皮细胞中的表达及意义

目的:动态观察结缔组织生长因子(CTGF)蛋白在血吸虫病肝纤维化鼠肝窦内皮细胞表达水平的时相变化,探讨其与肝窦内皮细胞下基底膜形成的关系。方法:采用腹部敷贴法建立血吸虫病肝纤维化模型,HE、Masson染色和透射电镜观察其病理变化;免疫组化技术检测CTGF、Ⅳ型胶原(ColⅣ)和层粘连蛋白(LN)在小鼠肝脏组织中的表达和分布;并应用彩色图像分析仪进行定量分析。结果:与正常对照组比较,模型组鼠肝窦内皮细胞表达CTGF蛋白阳性或强阳性,肝窦壁LN、ColⅣ表达水平增高,且随着肝纤维化的发展,CTGF和LN、ColⅣ表达逐渐增强,肝窦内皮下基底膜逐渐增厚。图像定量分析两组平均吸光度值、平均灰度值和阳性面积比具有统计学差异;CTGF蛋白与LN、ColⅣ水平呈正相关。结论:血吸虫病肝纤维化时小鼠肝窦内皮细胞通过CTGF蛋白表达上调,调控细胞外基质产生,导致ColⅣ、LN分泌增加,参与肝窦内皮下基底膜形成,从而引起肝内微循环障碍。     

Influence of Platelet-derived Growth Factor on Lens Epithelial Cell Proliferation and Differentiation

The expression pattern of platelet-derived growth factor (PDGF) and its receptor suggest a role in lens cell proliferation. PDGF is strongly expressed in the iris and ciliary body, situated opposite the proliferative cells of the lens epithelium which express the PDGF- &#102 receptor. In this study, using lens epithelial explant cultures, we report that PDGF can induce a dose and time dependent increase in lens cell DNA synthesis. Culturing lens explants with both PDGF and FGF (a mitogen and differentiation factor for lens cells) resulted in responses greater than those induced by either growth factor alone. PDGF did not induce any changes typical of fibre differentiation; however, in combination with FGF it potentiated the fibre differentiating activity of FGF. Results obtained in this study support previous indications that PDGF has an important role in regulating lens cell proliferation. In addition, PDGF may have a role in potentiating FGF-induced lens fibre differentiation in vivo.     

吡格列酮对高脂血症大鼠主动脉结缔组织生长因子表达水平的影响

目的:观察吡格列酮对高脂血症大鼠主动脉结缔组织生长因子(CTGF)表达的影响。方法:将26只雄性SD大鼠按体重随机分成普通饲料对照组(9只)和高脂饲料组(17只),分笼饲养,12周后检测其空腹血脂水平,以确定高脂饲料组造模成功;再将成模大鼠随机分为模型组(8只)和吡格列酮组(9只)。第13周起予吡格列酮组大鼠吡格列酮(10mg/kg/天),余两组用等量生理盐水连续灌胃4周后,平行检测各组大鼠血脂水平,主动脉组织形态学(包括光镜、电镜),免疫组化检测主动脉CTGF表达,并用Image-ProPlus图像分析系统分析染色结果。结果:高脂饲料喂养12周后,大鼠总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白胆固醇(LDL-C)水平较对照组明显升高(P<0.01),造模成功。吡格列酮组给药4周后血清TC、TG水平较模型组明显降低(P<0.01),主动脉CTGF表达也显著下调(P<0.01)。光镜下可见吡格列酮组内皮细胞偶有脱落,内膜下少量泡沫细胞,平滑肌细胞轻度增生;电镜下可见吡格列酮组大鼠血管内皮层整齐,偶见线粒体水肿,中膜平滑肌细胞少见泡沫样变,均较模型组好转。结论:主动脉CTGF表达增加参与了高脂血症对大鼠血管内膜的损伤,吡格列酮干预能够降低主动脉CTGF表达,减轻主动脉内膜损伤,对动脉粥样硬化有保护作用。     

Homocysteine Toxicity in Connective Tissue: Theories,Old and New

Hyperhomocysteinemia causes connective tissue pathology. Several theories on the mechanism of homocysteine toxicity in connective tissue are reviewed briefly. A possible new mechanism was revealed recently in the discovery of a reaction in which homocysteine thiolactone is converted to mercaptopropionaldehyde. The reaction is the Strecker degradation of amino acids in which ninhydrin is replaced by the structurally similar dehydroascorbic acid. The reaction may occur in vivo and may be pathogenic to connective tissue in four ways: (1) the reaction may deplete ascorbic acid that is required for collagen synthesis, (2) the mercaptoaldehyde product may interfere with collagen synthesis, (3) the mercaptoaldehyde may cause abnormal cross-linking of collagen molecules, and (4) the mercaptoaldehyde may attach to collagen molecules rendering them antigenic and triggering an autoimmune response.     

血管内皮生长因子和骨形态发生蛋白在骨组织工程中的作用

背景：在临床中由于骨质疏松性骨折、创伤、先天性骨发育不良、进行性骨骼紊乱引起的大段结构性骨缺损非常常见,组织工程骨为骨缺损的修复提供了新希望。骨组织工程研究中关于生长因子的研究较多,已取得了一些成果,如何在时间上控制各种不同生物活性的生长因子是研究热点。目的：综述血管内皮生长因子和骨形态发生蛋白在血管化组织工程骨中的研究进展。方法：由第一作者用计算机检索中国期刊全文数据库(CNKI：1990至2015年)Medline(1990至2015年)数据库,检索词分别为“成骨因子、成血管因子、组织工程骨、骨修复、血管化、血管内皮生长因子、骨形态发生蛋白、序贯释放、种子细胞、细胞支架”和“osteogenic factors,angiogenic factors,tissue engineering bone,bone repair,vascularization,VEGF,BMPs,sequential release,seed cells,cytoskeleton”,语言分别设定为中文和英文。检索有关血管内皮生长因子和骨形态发生蛋白在骨修复作用中的研究,并总结作用机制及研究方向。结果与结论：共检索到313篇文献,按纳入及排除标准筛选后,纳入文章87篇。结果表明,骨缺损的重建伴随着多种不同的生物活性分子,各自具有不同的潜能和效率。血管和成骨是骨修复两个最为重要的过程,成骨生长因子在维持骨骼结构和骨形成中发挥重要的作用,而成血管生长因子可以为组织的生长、分化和功能化提供氧气和营养物质,双因子或多因子联合作用是目前但骨组织工程中的一个发展方向,成骨与成血管生物因子比单独使用其中一种具有更好的成骨效果,但是控制外源性成骨与成血管生物因子的释放剂量是在治疗过程中的关键研究问题。中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

结缔组织生长因子siRNA对大鼠血管平滑肌细胞增殖和迁移的影响

目的观察结缔组织生长因子(CTGF)siRNA对CTGF表达及对大鼠血管平滑肌细胞(VSMC)增殖和迁移的影响。方法培养大鼠血管平滑肌细胞,用脂质体法将抗CTGF483siRNA进行转染,Western blot法检测转染效果,MTT法及划痕实验测定对VSMC增殖和迁移的影响。结果转染CTGF483siRNA组的CTGF表达明显低于未转染组,MTT法显示转染CTGF483siRNA后VSMCs增殖减慢,划痕试验证实转染CTGF483siRNA后VSMCs迁移受到抑制。结论CTGFsiRNA可以抑制VSMC增殖和迁移,有望为颈动脉粥样硬化性缺血性脑血管病的预防和治疗新靶点。     

冠心病患者血浆组织因子、组织因子途径抑制物水平及抗凝血酶-Ⅲ活性的研究

目的 :检测不同类型冠心病 (CHD)患者血浆组织因子 (TF)、组织因子途径抑制物 (TFPI)抗原水平及抗凝血酶 Ⅲ(AT Ⅲ)活性的变化并探讨其临床意义。方法 :TF抗原 (TF Ag)、TFPI抗原 (TFPI Ag)均采用双抗夹心ELISA法测定 ,AT Ⅲ活性采用发色底物法。结果 :急性心肌梗死 (AMI)组、不稳定型心绞痛 (UAP)组和稳定型心绞痛 (SAP)组TF抗原水平治疗前后均显著高于对照组 (P <0 .0 5 ) ,陈旧性心肌梗死(OMI)组治疗前后与对照组水平接近 (P >0 .0 5 ) ;各组治疗前后TFPI抗原水平均高于对照组 (P <0 .0 5 ) ;各组治疗前AT Ⅲ活性均显著低于对照组 (P <0 .0 5 ) ,治疗后 ,AMI组和UAP组AT Ⅲ活性升高至对照组水平 (P >0 .0 5 ) ,SAP组和OMI组仍低于对照组 (P <0 .0 5 )。结论 :AMI及心绞痛患者发作期 ,外源性凝血途径被启动 ,TF过度表达 ,TFPI反馈性增高 ,AT Ⅲ活性因被过多拮抗和消耗而降低 ,血液处于高凝状态。     

支气管哮喘患者血清中VEGF、 EGF等相关生长因子表达及与临床表型、肺功能以及病情程度相关性研究

目的 研究支气管哮喘患者血清中表皮生长因子(EGF)、碱性成纤维细胞生长因子(bFGF)、血小板衍生生长因子(PDGF)以及血管内皮生长因子(VEGF)的表达特征及其与BA发生、BA患者肺功能、病情程度以及临床表型间的相关性.方法 选取本院2016年1月至2017年1月间收治的72例支气管哮喘患者纳入研究组,以同期在本院接受健康体检的72例健康受检者作为对照组,对比两组各项生长因子的表达水平;分别根据临床表型、肺功能以及病情程度将研究组患者划分为相应亚组,对比各亚组间各项生长因子的表达水平,以明确支气管哮喘患者血清中各项相关生长因子的表达特征;分析各项生长因子表达水平与各亚组间的相关性.结果 研究组患者VEGF、EGF、bFGF、PDGF-AA、PDGF-BB表达水平与对照组受检者间差异均具有统计学意义,P＜0.05;研究组中嗜酸粒细胞表型患者的VEGF、EGF、bFGF、PDGF-AA、PDGF-BB表达水平与中性粒细胞表达患者差异均具有统计学意义,P＜0.05;研究组中FEV1＜50％患者的VEGF、EGF、bFGF、PDGF-AA、PDGF-BB表达水平与FEV1≥50％患者差异均具有统计学意义,P＜0.05;轻度亚组、中度亚组与重度亚组间VEGF、EGF、bFGF、PDGF-AA、PDGF-BB表达水平差异均具有统计学意义,P＜O.05.VEGF、EGF、PDGF-AA表达水平与BA疾病发生间具有高度相关性,P＜0.05;VEGF、EGF、bFGF、PDGF-AA、PDGF-BB与BA临床不同表型、肺功能及病情程度间均具有相关性,P＜0.05.结论 支气管哮喘患者血清中EGF、b FGF、PDGF、VEGF表达水平与健康者比较具有明显特征,并且各项生长因子的表达水平与患者的临床表型、肺功能以及病情程度具有明确的相关性.