微晶削磨术后瘢痕组织的形态学观察

目的：建立兔耳增生性瘢痕模型并观察微晶削磨术后瘢痕组织病理学的改变。方法：通过建立兔耳腹面瘢痕模型并对其进行微晶磨削处理，肉眼观察瘢痕的形态变化及其在光镜下成纤维细胞的含量和各类型胶原纤维的变化。结果：对兔耳增生性瘢痕进行微晶磨削术后瘢痕面积明显缩小,瘢痕组织表面鳞状上皮棘细胞层细胞增生，表皮下真皮乳头层及其真皮层内成纤维细胞轻度增生，胶原纤维排列趋于整齐，Ⅰ型胶原含量减少，而Ⅲ型胶原含量明显增多。结论：微晶磨削术对于皮肤浅表性瘢痕的修复与改建具有积极的意义。     

三七总甙对增生性瘢痕的作用

背景：增生性瘢痕的传统治疗手段如手术切除、类固醇激素、抗代谢药、免疫抑制剂和放射疗法等,均易复发或有严重的不良作用而限制了其临床应用。近年来应用活血化瘀类中药提取成分治疗增生性瘢痕取得了较为理想的进展,而且不良反应小。 目的：通过组织染色及相关基因检测的方法,观察三七总甙对兔耳增生性瘢痕的作用以及对转化生长因子β1 mRNA表达的影响。 方法：创建兔耳增生性瘢痕模型,约术后4周,按随机数字表法将24只大耳白兔分为3组,三七总甙组、曲安奈德组及空白对照组每组8只。以曲安奈德为阳性对照,分组分次瘢痕基底局部给药,用药5次后取瘢痕组织,以苦味酸酸性复红染色法、苏木精-伊红染色法对胶原纤维、成纤维细胞进行观察,应用反转录PCR方法检测细胞内转化生长因子β1基因的表达情况。 结果与结论：苏木精-伊红染色结果示三七总甙组兔耳瘢痕中成纤维细胞数量低于空白对照组,与曲安奈德组无明显差异。苦味酸酸性复红染色结果示三七总甙组兔耳瘢痕中胶原纤维较空白对照组排列整齐,与曲安奈德组无明显差异。基因检测结果显示三七总甙组、曲安奈德组转化生长因子β1 mRNA的表达显著低于空白对照组(P < 0.05)。提示三七总甙及曲安奈德均能通过抑制转化生长因子β1 mRNA的表达,降低转化生长因子β1在瘢痕组织中的含量,从而抑制成纤维细胞的过度增殖及以胶原为主的细胞外基质过度沉积,进而降低胶原纤维合成,来达到有效抑制瘢痕组织过度增生的目的。     

不同类型克山病心肌间质纤维化的形态学分析

目的揭示克山病心肌间质中胶原的类型、分布、排列与含量。方法采用本室克山病尸检材料，应用苦味酸－天狼星红染色和偏振光显微镜观察和图像分析技术，系统地研究了克山病心肌胶原的变化。结果在急性克山病的瘢痕灶内，胶原排列成疏网状，Ⅲ型胶原比例增加。慢性克山病的瘢痕中，胶原排列致密，Ⅰ型胶原比例增加，Ⅰ／Ⅲ型胶原比值增大。在心肌细胞非坏死区域胶原含量明显高于对照组，出现间质纤维化，以慢性克山病最明显。亚急性和慢性克山病的心内膜厚度和血管周围胶原体密度明显高于对照组。结论在克山病心肌细胞变性坏死的同时伴随着胶原代谢的紊乱，胶原蓄积，心肌间质胶原发生重构，是克山病心功能不全的重要因素之一。     

早期注射丹参酮ⅡA磺酸钠可抑制兔耳增生性瘢痕

背景：丹参酮ⅡA磺酸钠具有抗氧化、抑制纤维化等作用。 目的：观察丹参酮ⅡA磺酸钠早期局部注射对兔耳增生性瘢痕的影响。 方法：于兔耳腹侧面切除全层皮肤及软骨膜建立瘢痕模型,造模后28 d,分别在创面瘢痕内注射0.05,0.1,0.2 mg的丹参酮ⅡA磺酸钠或生理盐水,每周注射1次,共3周。 结果与结论：丹参酮ⅡA磺酸钠注射3周,兔耳瘢痕增生较轻,瘢痕厚度变薄;苏木精-伊红染色见瘢痕组织中胶原纤维减少,血管数目减少;Masson染色检测见瘢痕组织胶原纤维分布面积减少;电镜下瘢痕组织成纤维细胞数量减少,体积相对变小。说明早期注射丹参酮ⅡA磺酸钠对兔耳瘢痕增生有抑制作用。     

不同类型克山病心肌间纤维化的形态学分析

目的 揭示克山病心肌间质中胶原的类型、分布、排列与含量。方法 采用本室克山病尸检材料,应用苦味酸－天狼星红染色和偏振兴显微镜观察和图像分析技术,系统地研究了克山病心肌胶原的变化。结果 在急性克山病的瘢痕灶内,胶原排列成疏网状,Ⅲ型胶原比例增加。慢性克山病的瘢痕中,胶原排列致密,１胶原比例增中Ⅰ／Ⅲ型腾的比值增大,在心肌晨坏死区域胶原一明显高于对照组,出现间质纤维化,以慢性克山病最明显。亚急性和慢性     

自发性高血压大鼠心肌胶原网络的重建

目的 :观察在高血压不同时期心肌胶原网络的改变 ,为高血压的治疗及心脏功能的早期保护提供基础研究的依据。方法 :采用Masson三色法染色光镜观察和天狼星红 苦味酸饱和液染色偏振光显微镜观察两种方法 ,对 8周、1 2周、2 4周自发性高血压大鼠心肌胶原进行观察 ,采用计算机图象分析方法 ,对高血压不同时期心肌胶原总量和Ⅰ /Ⅲ型胶原比例的改变进行观测。结果 :1 2周时心肌胶原总量较 8周时增多 ,Ⅰ /Ⅲ型胶原量比例有轻度增加。2 4周时 ,心肌胶原增生明显 ,局部出现心肌纤维化 ,胶原网络构筑杂乱无章。血管周围胶原增生更为显著 ,且向周围心肌间质中大面积蔓延。Ⅰ型胶原增多 ,Ⅰ /Ⅲ型胶原量比例增大。结论 :在高血压时 ,心肌胶原网络重建 ,与病程关系密切 ,病程越长 ,胶原网络破坏越严重     

负极性驻极体5氟尿嘧啶贴剂抑制兔耳增生性瘢痕的生长

目的 :探讨不同干预手段对兔耳增生性瘢痕的生长影响。方法 :负极性驻极体、5-氟尿嘧啶(5-Fu)以及联合负极性驻极体与5-Fu分别作用于兔耳创面,4周后检测瘢痕增生指数,H-E染色和免疫组织化学显色检测转化生长因子-β(TGF-β)、Ⅰ型和Ⅲ型胶原的平均光密度。结果 :负极性驻极体、负极性驻极体与5-Fu联合均可通过抑制TGF-β的表达抑制Ⅰ型和Ⅲ型胶原的合成;且均可通过影响成纤维细胞的密度、胶原的合成和胶原纤维束的排布,抑制创面形成增生性瘢痕。结论 :负极性驻极体可影响胶原束的排布,且通过抑制TGF-β的表达抑制胶原的合成。驻极体和5-Fu联用作用机制与驻极体类似,但抑制瘢痕形成的效果更佳。     

绝经后骨质疏松症病人Ⅰ型胶原的变化

目的　研究Ⅰ型胶原合成及代谢在绝经后骨质疏松症发病中的作用。　方法　以绝经后骨质疏松性骨折病人股骨头置换术弃去的股骨头为样本 ;采用苦味酸天狼星红 偏振光及Ⅰ型胶原和基质金属蛋白酶 1(MMP 1)抗体免疫组织化学染色 ,从蛋白水平观察骨中Ⅰ型胶原的分布和量的变化 ;地高辛标记的Ⅰ型胶原和MMP 1cDNA基因探针进行原位杂交 ,从mRNA水平观察Ⅰ型胶原和MMP 1的分布和量的变化 ;从骨中提取Ⅰ型胶原测定肽链的羟基化水平 ,观察Ⅰ型胶原羟基化程度的改变。　结果　绝经后骨质疏松性骨折病人胶原含量降低 ,Ⅰ型胶原蛋白纤维变细、断裂 ,结构紊乱。而细胞内Ⅰ型胶原和MMP 1mRNA以及MMP 1蛋白水平均升高。Ⅰ型胶原蛋白赖氨酸羟基化程度升高而脯氨酸羟基化程度不变。　结论　Ⅰ型胶原合成及代谢在质和量上的改变在绝经后骨质疏松症发病中起重要作用。     

小鼠间质胶原和基膜成分在诱导排卵过程中的变化

目的　研究间质胶原和基膜成分在诱导排卵过程中的变化。　方法　苦味酸 -天狼星红偏振光法 ,生物素 -抗生物素蛋白间接免疫荧光 ,原位杂交。　结果　排卵前后 , 型胶原、 型胶原、层粘连蛋白的分布、数量 ,以及 型胶原和间质金属蛋白酶 - 2 m RNA的表达有一定改变。　结论　间质胶原和基膜成分对排卵过程有一定调控作用。     

miR-2116-3p在人增生性瘢痕中的表达及作用

背景：目前多项研究证实了miRNA影响增生性瘢痕的发生发展,Ⅰ型胶原与增生性瘢痕成纤维细胞密切相关,猜测miR-2116-3p也有可能与增生性瘢痕的发生发展有关系。目的：探讨miR-2116-3p在人增生性瘢痕中的表达及作用。方法：收集新疆医科大学第一附属医院6例患者的增生性瘢痕组织与6例患者重睑术后正常皮肤组织,采用qRT-PCR法检测miR-2116-3p与Ⅰ型胶原的表达。取增生性瘢痕组织,原代培养第3-6代成纤维细胞,分为阴性对照组、miR-2116-3p模拟物组和miR-2116-3p抑制物组,分别转染对应的序列,用CCK-8法与EdU试剂盒检测细胞增殖活力,划痕实验检测细胞迁移能力,流式细胞术检测细胞凋亡,qRT-PCR法与Western blot法检测Ⅰ型胶原、Ⅲ型胶原和α平滑肌肌动蛋白的基因与蛋白表达,双荧光素酶实验验证其靶向结合。结果与结论：(1)增生性瘢痕组织中miR-2116-3p的mRNA表达量低于正常皮肤组织(P <0.01),Ⅰ型胶原的mRNA表达量低于正常皮肤组织(P <0.01);(2)转染后24,48,72 h,与阴性对照组比较,miR-21...     

Adenosine promotes wound healing and mediates angiogenesis in response to tissue injury via occupancy of A(2A) receptors

Recent evidence indicates that topical application of adenosine A(2A) receptor agonists, unlike growth factors, increases the rate at which wounds close in normal animals and promotes wound healing in diabetic animals as well as growth factors, yet neither the specific adenosine receptor involved nor the mechanism(s) by which adenosine receptor occupancy promotes wound healing have been fully established. To determine which adenosine receptor is involved and whether adenosine receptor-mediated stimulation of angiogenesis plays a role in promotion of wound closure we compared the effect of topical application of the adenosine receptor agonist CGS-21680 (2-p-[2-carboxyethyl]phenethyl-amino-5'-N-ethylcarboxamido-adenosine) on wound closure and angiogenesis in adenosine A(2A) receptor knockout mice and their wild-type littermates. There was no change in the rate of wound closure in the A(2A) receptor knockout mice compared to their wild-type littermates although granulation tissue formation was nonhomogeneous and there seemed to be greater inflammation at the base of the wound. Topical application of CGS-21680 increased the rate of wound closure and increased the number of microvessels in the wounds of wild-type mice but did not affect the rate of wound closure in A(2A) receptor knockout mice. Similarly, in a model of internal trauma and repair (murine air pouch model), endogenously produced adenosine released into areas of internal tissue injury stimulates angiogenesis because there was a marked reduction in blood vessels in the walls of healing air pouches of A(2A) receptor knockout mice compared to their wild-type controls. Inflammatory vascular leakage and leukocyte accumulation in the inflamed air pouch were similarly reduced in the A(2A) receptor knockout mice reflecting the reduced vascularity. Thus, targeting the adenosine A(2A) receptor is a novel approach to promoting wound healing and angiogenesis in normal individuals and those suffering from chronic wounds.     

Adenosine A2A, but not A1, receptors mediate the arousal effect of caffeine

Caffeine, a component of tea, coffee and cola, induces wakefulness. It binds to adenosine A1 and A2A receptors as an antagonist, but the receptor subtype mediating caffeine-induced wakefulness remains unclear. Here we report that caffeine at 5, 10 and 15 mg kg(-1) increased wakefulness in both wild-type mice and A1 receptor knockout mice, but not in A2A receptor knockout mice. Thus, caffeine-induced wakefulness depends on adenosine A2A receptors.     

胶原蛋白具有促进肿瘤相关巨噬细胞向M1型极化的作用

目的 以临床结直肠肿瘤患者标本和原位癌小鼠模型以及小鼠腹腔巨噬细胞为研究对象,探讨结直肠肿瘤微环境中胶原蛋白促进肿瘤相关巨噬细胞极化的作用。方法 应用免疫荧光染色检测结直肠癌患者肿瘤组织巨噬细胞的分布,检测原位癌模型小鼠肿瘤组织巨噬细胞的量和分布;Masson染色肿瘤组织内胶原纤维。提取小鼠腹腔巨噬细胞,接种于不同浓度Ⅰ型胶原蛋白包被的培养皿,Real-time PCR 检测巨噬细胞的肿瘤坏死因子（TNF）、诱导型一氧化氮合酶（iNOS）、CD163、CD206等基因表达。 结果 人结肠癌组织标本和小鼠原位癌模型可见巨噬细胞主要分布于肿瘤组织的间质,主要为M1型,肿瘤实质部分巨噬细胞主要为M2型;细胞实验表明,胶原蛋白明显促进巨噬细胞向促进炎症抑制肿瘤的M1型巨噬细胞极化,抑制巨噬细胞向促进肿瘤的M2型极化;Masson染色可见间质内含有大量胶原纤维,而肿瘤实质细胞周围胶原纤维较少。 结论 胶原蛋白具有促进巨噬细胞向M1型极化的作用,而肿瘤实质细胞周围胶原纤维减少,可能与巨噬细胞向M2型极化,发挥促进肿瘤发生发展的作用有关。     

增生性瘢痕中时序变化的骨形态发生蛋白及其受体

背景：骨形态发生蛋白与瘢痕的形成和发展相关,而成纤维细胞与瘢痕的增殖、成熟关系密切,但骨形态发生蛋白在不同时期增生性瘢痕成纤维细胞中的表达变化尚少见报道。 目的：检测骨形态发生蛋白2/4、骨形态发生蛋白7、骨形态发生蛋白IA型受体在不同时期增生性瘢痕成纤维细胞中的表达水平。 方法：应用免疫组织化SP法检测来自2008年3月至2010年7月广西壮族自治区人民医院美容整形外科患者的增殖期增生性瘢痕和成熟期增生性瘢痕中成纤维细胞中骨形态发生蛋白2/4、骨形态发生蛋白7、骨形态发生蛋白IA型受体的表达。 结果与结论：增生性瘢痕增殖期成纤维细胞中骨形态发生蛋白2/4的表达水平显著高于成熟期(P < 0.05);但不同时期增生性瘢痕成纤维细胞中骨形态发生蛋白7和骨形态发生蛋白IA型受体的表达差异无显著性意义  (P > 0.05)。表明在增生性瘢痕由增生期至成熟期的发展过程中,其成纤维细胞中的骨形态发生蛋白2/4的表达下调,而骨形态发生蛋白7、骨形态发生蛋白IA型受体的表达无变化。 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;组织工程全文链接：     

黄韧带退变与腰椎管狭窄症椎管形态的变化

背景：目前研究发现Ⅰ型及Ⅱ型胶原在结缔组织的增殖与分化、软骨的形成和吸收等过程中起十分重要的作用。 目的：探讨腰椎黄韧带退变与腰椎管狭窄症椎管形态变化的相关性。 方法：收集中央型腰椎管狭窄症36例和外伤性腰椎骨折20例患者的黄韧带标本分别为实验组和对照组,采用酶联免疫吸附法测定Ⅰ型和Ⅱ型胶原;CT测量腰椎硬膜横截面积;黄韧带行苏木精-伊红和Masson染色,观察组织病理学变化。 结果与结论：实验组黄韧带厚度、Ⅰ型及Ⅱ型胶原含量高于对照组,腰椎硬膜横截面积和Ⅰ型/Ⅱ型比值低于对照组(P <　0.05);病理学显示实验组黄韧带弹性纤维排列紊乱,数量减少,胶原纤维增生。提示腰椎黄韧带Ⅰ型、Ⅱ型胶原含量及Ⅰ型/Ⅱ型比值发生改变,可能引起黄韧带厚度增加,导致椎管横截面积减小,参与腰椎管狭窄症的发生。关键词：黄韧带;腰椎管狭窄症;Ⅰ型胶原;Ⅱ型胶原;病理学 型/II型比值 doi:10.3969/j.issn.1673-8225.2012.17.001     

细菌纤维素减轻兔耳增生性瘢痕的作用

背景：国内外已有研究报道细菌纤维素对皮肤创伤愈合具有促进作用,但是其对增生性瘢痕是否有治疗作用尚不清楚。 目的：观察细菌纤维素对兔耳增生性瘢痕的疗效。 方法：建立兔耳腹侧增生性瘢痕模型,术后第21天创面上皮化后,对每只兔耳5个不同瘢痕面随机给予5种不同处理方式：持水性分别为1∶5,1∶6,1∶8细菌纤维素组、阳性对照组(贴敷瘢痕贴)、阴性对照组(未贴任何敷料且瘢痕自然生长)。观察不同处理后第0,14,21,28,42,56天瘢痕面大体形态学及组织学变化。 结果与结论：持水性1∶5,1∶6,1∶8细菌纤维素组瘢痕增生厚度低于阴性对照组,但高于阳性对照组(P < 0.01)。与阴性对照组比较,持水性1∶5,1∶6,1∶8细菌纤维素组瘢痕组织中真皮层薄,成纤维细胞少,胶原纤维较细、排列较整齐;与阳性对照组组比较,持水性1∶5,1∶6,1∶8细菌纤维素组成纤维细胞数稍多,胶原也稍粗、排列也稍不整齐。3种细菌纤维素组间瘢痕厚度及成纤维细胞数量为1∶5细菌纤维素组> 1∶6细菌纤维素组> 1∶8细菌纤维素组(P  < 0.05)。说明细菌纤维素有效抑制了兔耳创面愈合后增生性瘢痕的形成,并且持水性越高,效果越好。     

Morphological and immunochemical differences between keloid and hypertrophic scar.

There are two types of excessive scarring, keloid and hypertrophic scar. Contrary to hypertrophic scars, keloids do not regress with time, are difficult to revise surgically, and do not provoke scar contractures. These two lesions require different therapeutic approaches but are often confused because of an apparent lack of morphological differences. We have investigated the collagen organization and the possible presence of alpha-smooth muscle (SM) actin-expressing myofibroblasts in these conditions. Keloids contain large, thick collagen fibers composed of numerous fibrils closely packed together. In contrast hypertrophic scars exhibit modular structures in which fibroblastic cells, small vessels, and fine, randomly organized collagen fibers are present. We confirm that such nodular structures are always present in hypertrophic scar and rarely in keloid. Furthermore, only nodules of hypertrophic scars contain alpha-SM actin-expressing myofibroblasts. Electron microscopic examination supports the above-mentioned differences in collagen organization and in fibroblastic features and shows the presence of an amorphous extracellular material surrounding fibroblastic cells in keloid. The presence in hypertrophic scar myofibroblasts of alpha-SM actin, the actin isoform typical of vascular SM cells, may represent an important element in the pathogenesis of contraction. Interestingly, when placed in culture fibroblasts from hypertrophic scars and keloid express similar amounts of alpha-SM actin, suggesting that local microenvironmental factors influence in vivo the expression of this protein. Thus several morphological and immunohistochemical differences exist between hypertrophic scar and keloid that are useful for the biological and pathological characterization of the two lesions.     

Pharmacological blockade of A2A receptors prevents dermal fibrosis in a model of elevated tissue adenosine

Adenosine is a potent modulator of inflammation and tissue repair. We have recently reported that activation of adenosine A(2A) receptors promotes collagen synthesis by human dermal fibroblasts and that blockade or deletion of this receptor in mice protects against bleomycin-induced dermal fibrosis, a murine model of scleroderma. Adenosine deaminase (ADA) is the principal catabolic enzyme for adenosine in vivo, and its deficiency leads to the spontaneous development of pulmonary fibrosis in mice. The aim of this study was to characterize further the contributions of endogenous adenosine and adenosine A(2A) receptors to skin fibrosis. Taking advantage of genetically modified ADA-deficient mice, we herein report a direct fibrogenic effect of adenosine on the skin, in which increased collagen deposition is accompanied by increased levels of key mediators of fibrosis, including transforming growth factor beta1, connective tissue growth factor, and interleukin-13. Pharmacological treatment of ADA-deficient mice with the A(2A) receptor antagonist ZM-241385 prevented the development of dermal fibrosis in this model of elevated tissue adenosine, by reducing dermal collagen content and expression of profibrotic cytokines and growth factors. These data confirm a fibrogenic role for adenosine in the skin and reveal A(2A) receptor antagonists as novel therapeutic agents for the modulation of dermal fibrotic disorders.     

Selective attenuation of psychostimulant-induced behavioral responses in mice lacking A(2A) adenosine receptors

A(2A) adenosine receptors are highly expressed in the striatum where they modulate dopaminergic activity. The role of A(2A) receptors in psychostimulant action is less well understood because of the lack of A(2A)-selective compounds with access to the central nervous system. To investigate the A(2A) adenosinergic regulation of psychostimulant responses, we examined the consequences of genetic deletion of A(2A) receptors on psychostimulant-induced behavioral responses. The extent of dopaminergic innervation and expression of dopamine receptors in the striatum were indistinguishable between A(2A) receptor knockout and wild-type mice. However, locomotor responses to amphetamine and cocaine were attenuated in A(2A) knockout mice. In contrast, D(1)-like receptor agonists SKF81297 and SKF38393 produced identical locomotor stimulation and grooming, respectively, in wild-type and A(2A) knockout mice. Similarly, the D(2)-like agonist quinpirole produced motor-depression and stereotypy that were indistinguishable between A(2A) knockout and wild-type mice. Furthermore, attenuated amphetamine- (but not SKF81297-) induced locomotion was observed in pure 129-Steel as well as hybrid 129-SteelxC57BL/6 mice, confirming A(2A) receptor deficiency (and not genetic background) as the cause of the blunted psychostimulant responses in A(2A) knockout mice.These results demonstrate that A(2A) receptor deficiency selectively attenuates psychostimulant-induced behavioral responses and support an important role for the A(2A) receptor in modulating psychostimulant effects.     

Genetic interdependence of adenosine and dopamine receptors: evidence from receptor knockout mice

Dopamine and adenosine receptors are known to share a considerable overlap in their regional distribution, being especially rich in the basal ganglia. Dopamine and adenosine receptors have been demonstrated to exhibit a parallel distribution on certain neuronal populations, and even when not directly co-localized, relationships (both antagonistic and synergistic) have been described. This study was designed to investigate dopaminergic and purinergic systems in mice with ablations of individual dopamine or adenosine receptors. In situ hybridization histochemistry and autoradiography was used to examine the level of mRNA and protein expression of specific receptors and transporters in dopaminergic pathways. Expression of the mRNA encoding the dopamine D2 receptor was elevated in the caudate putamen of D1, D3 and A2A receptor knockout mice; this was mirrored by an increase in D2 receptor protein in D1 and D3 receptor knockout mice, but not in A2A knockout mice. Dopamine D1 receptor binding was decreased in the caudate putamen, nucleus accumbens, olfactory tubercle and ventral pallidum of D2 receptor knockout mice. In substantia nigra pars compacta, dopamine transporter mRNA expression was dramatically decreased in D3 receptor knockout mice, but elevated in A2A receptor knockout mice. All dopamine receptor knockout mice examined exhibited increased A2A receptor binding in the caudate putamen, nucleus accumbens and olfactory tubercle. These data are consistent with the existence of functional interactions between dopaminergic and purinergic systems in these reward and motor-related brain regions.