瘢痕疙瘩发病机理的研究进展

目的综述瘢痕疙瘩发病机理的研究进展。方法广泛查阅近年来有关瘢痕疙瘩发生的组织学、病理学、生理学、免疫学和遗传学方面的相关文献，并作综合分析。结果经查阅发现。最近的基因组扫描首次提供了瘢痕疙瘩易感基因位点在染色体2q23和7p11的遗传学证据。结论识别瘢痕疙瘩易感基因将显著提高学者们对瘢痕疙瘩生物学发病机制的认识，这不仅有利于瘢痕疙瘩的诊断和治疗，而且有助于对创伤愈合机理产生新的认识。     

福建省两个汉族瘢痕疙瘩家系易感基因位点与染色体7p11的连锁分析

目的：探讨福建省两个汉族瘢痕疙瘩家系易感基因位点是否与7p11存在连锁关系。方法：从来自福建省2个汉族瘢痕疙瘩家系中选出26名具有较高遗传学研究意义的成员作为研究对象,采集他们的外周静脉血样,提取基因组DNA,参照国外最近相似研究的方法,在染色体7p11上选取已知的4个最大两点LOD值的微卫星为遗传标记,经PCR扩增,产物基因分型,再进行连锁分析。结果：在重组率Θ=0～0．1时,这些微卫星标记的两点LOD值都小于-2,排除这些标记与染色体7p11的连锁关系。结论：本研究发现福建省两个汉族瘢痕疙瘩家系的易感基因位点不在染色体7p11上的遗传学证据,说明瘢痕疙瘩易感基因位点存在异质性。     

福建省两个汉族瘢痕疙瘩家系易感基因位点与染色体2q23的连锁分析

目的：探讨福建省两个汉族瘢痕疙瘩家系易感基因位点是否与2q23存在连锁关系。方法：从来自福建省2个汉族瘢痕疙瘩家系中选出26名具有较高遗传学研究意义的成员作为研究对象,采集他们的外周静脉血样,提取基因组DNA,参照国外最近相似研究的方法,在染色体2q23上选取已知的6个最大两点LOD值的微卫星为遗传标记,经PCR扩增,产物基因分型,再进行连锁分析。结果：在重组率θ=0时,这些微卫星标记的两点LOD值都小于-2;在重组率θ=0.05时,它们的两点LOD值均小于-1;可以否定这些标记与2q23的连锁关系。结论：本研究发现福建省两个汉族瘢痕疙瘩家系的易感基因位点不在染色体2q23上的遗传学证据,说明瘢痕疙瘩易感基因位点存在异质性。     

中国汉族一瘢痕疙瘩家系易感基因的定位研究

目的定位中国汉族瘢痕疙瘩家系的易感基因。方法采集1个5代发病的中国汉族瘢痕疙瘩大家系32名成员的外周静脉血样,提取基因组DNA;设定Fas基因为导致该家系发病的一个候选基因,选取位于10q23.31上Fas基因周围共约10Mbp范围内与细胞凋亡障碍或肿瘤发生有关的所有已知基因相邻的微卫星标记D10S1687、D10S1765、D10S1735和D10S1562共4个,对这些微卫星位点进行PCR扩增,产物片断基因分型和连锁分析。结果连锁分析发现微卫星标记D10S1765LODZMAX为1.74,D10S1735LODZMAX为1.51,支持连锁;D10S1562LODZMAX为0.59,不排除连锁;在θ=0.0～0.10时,D10S1687标记的所有LOD值都小于-2,排除连锁。结论我们的研究首次发现了该中国汉族瘢痕疙瘩家系的易感基因可能位于10q23.31上D10S1765与D10S1735两位点间约1Mbp区域的遗传学证据。     

中国人群瘢痕疙瘩家系与染色体 2q23 和 7p11 的连锁分析

目的探讨中国人群瘢痕疙瘩家系是否与2q23和7p11存在连锁关系。方法选择两个中国人群瘢痕疙瘩大家系,从中共选出51名成员,采集其外周静脉血样,提取基因组DNA;参照国外最近相似研究的文献报道,在染色体2q23和7p11上,分别选取6个和4个微卫星标记,经多重PCR扩增,产物片断基因分型,再进行连锁分析。结果在重组率θ=0时,这些微卫星标记的两点LOD值绝大部分都小于-2,排除连锁关系存在。结论本研究首次发现了中国人群瘢痕疙瘩家系的易感基因位点不在染色体2q23和7p11上的遗传学证据,说明瘢痕疙瘩易感基因位点存在异质性。     

中国人群瘢痕疙瘩家系与染色体 2q23 和 7p11 的连锁分析

目的 探讨中国人群瘢痕疙瘩家系是否与2q23和7p11存在连锁关系.方法 选择两个中国人群瘢痕疙瘩大家系,从中共选出51名成员,采集其外周静脉血样,提取基因组DNA;参照国外最近相似研究的文献报道,在染色体2q23和7p11上,分别选取6个和4个微卫星标记,经多重PCR扩增,产物片断基因分型,再进行连锁分析.结果 在重组率θ=0时,这些微卫星标记的两点LOD值绝大部分都小于-2,排除连锁关系存在.结论 本研究首次发现了中国人群瘢痕疙瘩家系的易感基因位点不在染色体2q23和7p11上的遗传学证据,说明瘢痕疙瘩易感基因位点存在异质性.     

中国汉族瘢痕疙瘩家系易感基因位点的定位分析研究

目的 定位中国汉族瘢痕疙瘩家系的易感基因位点.方法 采集2个4代发病的中国汉族瘢痕疙瘩大家系51例成员的外周静脉血样.提取基因组DNA;假定Fas基因为该家系致病基因的候选基因位点.选取位于10q23.31上Fas基因周围共约10Mbp范围内与细胞凋亡障碍有关的已知基因相邻的微卫星标记D10S1687、D10S1765、D10S1735和D10S1562,对这些微卫星位点进行PCR扩增,产物片断基因分型,再进行连锁分析.结果 在重组率θ=0～0.5时,这些微卫星标记的两点LOD值绝大部分都小于1,排除连锁关系存在.结论 研究发现中国汉族瘢痕疙瘩家系易感基因位点不在染色体10q23.31区域.     

瘢痕疙瘩成纤维细胞的基因组学研究

目的 寻找瘢痕疙瘩致病相关基因，探讨瘢痕疙瘩的发生机理。方法 利用含1100个人类肿瘤相关基因的cDNA芯片(cDNA—microarray)对耳垂和胸部瘢痕疙瘩及正常皮肤成纤维细胞进行检测，初步分析瘢痕疙瘩成纤维细胞与正常皮肤成纤维细胞基因总体表达的差异，并筛选出差异基因。结果 在耳垂及胸部瘢痕疙瘩成纤维细胞中，分别有8种和17种特异性表达基因被检出。在正常皮肤中特异性表达的细胞增殖抑制基因Mda-7，在耳垂及胸部瘢痕疙瘩成纤维细胞中均未被表达。结论 多种基因参与了瘢痕疙瘩的形成过程，瘢痕疙瘩成纤维细胞与正常皮肤成纤维细胞之间存在基因表达的差异，增殖因子受体PAR-1和增殖抑制基因Mda-7可能参与瘢痕疙瘩的形成。     

福建省两个汉族瘢痕疙瘩家系易感基因的定位研究

目的：开展汉族瘢痕疙瘩家系易感基因定位研究。方法：收集来自福建省不同地区的两个汉族瘢痕疙瘩家系，分别命名为A和B家系，从中选出26名具有较高遗传学研究意义的成员作为研究对象，采集他们的外周静脉血样，提取基因组DNA；设定Fas基因为导致家系发病的一个候选基因，选取位于10q23．31上Fas基因周围共约10Mbp范围内与细胞凋亡障碍或肿瘤发生有关的所有已知基因相邻的微卫星标记D10S1687、D10S1765、D10S1735和D10S1562共4个，对这些微卫星位点进行PCR扩增，产物片断基因分型和连锁分析。结果：A家系微卫星标记D10S1765LODZMAX为1．86，D10S1735LODZMAX为1．29，支持连锁；D10S1562LODZMAX为0．42，不排除连锁；在O=0．0～0．10时，D10S1687标记的所有LOD值都小于-2，排除连锁。B家系微卫星标记D10S1765LODZMAX为1．63，D10S1735LODM为1．37，支持连锁；D10S1562LODZMAX为0．25，不排除连锁；在0=0．0～0．10时，D10S1687标记的所有LOD值都小于-2，排除连锁。结论：本研究提示这两个来自福建的汉族瘢痕疙瘩家系的易感基因可能位于10q23．31上D10S1765与D10S1735两位点间约1Mbp区域。     

p53基因第72位密码子多态与瘢痕疙瘩易感性研究

目的 探讨p53基因第72位密码子多态与瘢痕疙瘩遗传易感性的关系。方法 采用聚合酶链反应-反向斑点杂交、DNA直接测序方法，检测了15例瘢痕疙瘩患者与15名正常人对照的p53基因第72位密码子的基因型。结果 瘢痕疙瘩组的Pro等位基因及Pro／Pro基因型频率明显高于对照组(P值分别为0．035、0．030)，Pro／Arg、Arg／Arg基因型频率在病例组和对照组的分布差异无显著性。结论 p53基因第72位密码子Pro等位基因及Pro／Pro基因型可能是瘢痕疙瘩的易感因素。     

Macrophages of the M1 and M2 types play a role in keloids pathogenesis

The pathophysiology of keloid formation is unknown, however, macrophages are thought to play a role in keloid formation. Understanding the mechanism(s) of keloid development might be crucial in developing a new treatment regimen for keloids. The aim of this study was to understand possible status of M1 and M2 type macrophages in the pathogenesis of keloid. Thirty cases of Keloid tissues were selected according to our inclusion and exclusion criteria, as well as 30 normal scars, were enrolled in our study as a control group. An excisional biopsy was harvested and ELISA was done on keloid tissue and normal scar samples, with CD68, the surface marker for M1 and CD163 representing M2. The results revealed the low expression of M1 (CD68) in keloid tissue meanwhile high levels of M1 were detected in normal scars. We also detected that higher tissue expression of M2 (CD163) was significantly associated with keloid cases when compared to low M2 expression in the control group. An important finding that was discovered during our study is that the M1 and M2 are significant predictors of keloid. Every increase of 1 ng/mL in M1 decreases the risk of keloid by 0.99 while every increase of one unit in M2 increases the risk of keloid by 2.01. This study concluded that the keloid formation could be a result of an abnormal response to tissue injury where there is an excessive entry of inflammatory cells into the wound, including macrophages and that the keloid incidence might be related to a decrease in M1 and an increase in M2.     

细胞外信号调节激酶及应激活化蛋白激酶通路蛋白在瘢痕疙瘩成纤维细胞中的表达

目的通过比较瘢痕疙瘩及正常皮肤中细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)、应激活化蛋白激酶(c-Jun amino-terminal kinase,JNK)信号通路的表达,探讨其在瘢痕疙瘩形成中的作用。方法取四川大学华西医院烧伤整形科收治的26例患者皮肤组织,其中行瘢痕疙瘩切除患者(实验组)16例,瘢痕疙瘩形成时间8个月～10年;胸部6例,耳垂4例,会阴部2例,肩部3例,腹部1例;均经病理检查确诊为瘢痕疙瘩。10例整形手术患者自愿捐赠的正常皮肤作为对照组,腹部4例,大腿3例,肩部2例,背部1例。取标本采用Envision二步法行免疫组织化学染色,观察磷酸化及非磷酸化JNK和ERK表达情况,并采用Image Pro Plus 4.5图像分析系统测定积分吸光度(IA)值,观察阳性染色强度。结果免疫组织化学染色观察显示,对照组正常皮肤成纤维细胞中未见明显的磷酸化及非磷酸化ERK、JNK阳性表达;而实验组主要在成纤维细胞中表达,阳性颗粒主要位于细胞质和细胞核内。实验组磷酸化ERK及JNK的IA值明显高于对照组(P<0.05),非磷酸化ERK及JNK两组间差异无统计学意义(P>0.05)。结论磷酸化JNK、ERK信号通路蛋白在瘢痕疙瘩中异常高表达,提示该通路可能与瘢痕疙瘩形成密切相关。     

瘢痕疙瘩成纤维细胞低密度培养的克隆能力分析

目的：比较瘢痕疙瘩成纤维细胞（KFs）和正常真皮成纤维细胞（NFs）间克隆形成能力的差异,探讨瘢痕疙瘩中病理性干细胞是否存在,及其对瘢痕疙瘩发生发展的影响。方法利用酶消化法获得瘢痕疙瘩和正常皮肤组织的原代细胞,以4000个/皿的密度接种,进行低密度培养,2周后观察细胞克隆的形成及形态变化。结果低密度培养条件下的瘢痕疙瘩成纤维细胞和正常皮肤成纤维细胞都可形成克隆,但KFs可形成明显的克隆集落,NFs形成的克隆不明显且松散。KFs克隆形成率高于NFs,为（0.80±0.21）%,而NFs为（0.18±0.06）%,两者间差异显著（P〈0.05）。结论低密度培养条件下,瘢痕疙瘩成纤维细胞的克隆形成能力高于正常皮肤成纤维细胞,可能与瘢痕疙瘩组织中存在病理性瘢痕疙瘩干细胞有关。     

A role for vitamin D and the vitamin D receptor in keloid disorder

Keloids are disfiguring fibroproliferative lesions that can occur in susceptible individuals following any skin injury. They are extremely challenging to treat, with relatively low response rates to current therapies and high rates of recurrence after treatment. Although several distinct genetic loci have been associated with keloid formation in different populations, there has been no single causative gene yet identified and the molecular mechanisms guiding keloid development are incompletely understood. Further, although it is well known that keloids are more commonly observed in populations with dark skin pigmentation, the basis for increased keloid risk in skin of colour is not yet known. Because individuals with dark skin pigmentation are at higher risk for vitamin D deficiency, the role of vitamin D in keloid pathology has gained interest in the keloid research community. A limited number of studies have found lower serum vitamin D levels in patients with keloids, and reduced expression of the vitamin D receptor (VDR) in keloid lesions compared with uninjured skin. Vitamin D has documented anti-inflammatory, anti-proliferative and pro-differentiation activities, suggesting it may have a therapeutic role in suppression of keloid fibrosis. Here we review the evidence supporting a role for vitamin D and VDR in keloid pathology.     

A wound contraction experimental model for studying keloids and wound-healing modulators

Preventing and treating hypertrophic and keloid scars is difficult because of the lack of knowledge about their genesis. Tissue repair can be studied with biocompatible matrices and ex vivo cultures of different cell types. We used an experimental model where collagen gels populated by human fibroblasts underwent progressive contraction, allowing the study of wound healing remodeling. The fibroblast-populated lattices showed the greater contraction of the gel populated by fibroblasts from keloids versus fibroblasts from normal skin. Moreover, fibroblast growth factor (FGF) and transforming growth factor beta (TGF-beta) involved in scar formation were added to the collagen gels populated by normal skin fibroblasts. TGF-beta caused an increase in gel contraction; FGF did not. The mean percentages of contraction of the gels populated by keloid fibroblasts were very similar to the percentages of gels populated by normal skin fibroblasts with added TGF-beta. These observations confirm the existing hypothesis that TGF-beta may be involved in keloid formation.     

Gene expression patterns in isolated keloid fibroblasts

Keloid scars after skin trauma are a significant clinical problem, especially in black populations, in which the incidence of keloids has been estimated at 4-16%. Keloids are abnormal dermal proliferative scars secondary to dysregulated wound healing. Despite several biochemical studies on the role of extracellular matrix proteins and growth factors during keloid formation, we still do not know what molecules and signals induce this change. Fibroblasts are thought to be the major inductive cell for keloid scar formation. The aim of this study was to identify gene expression patterns that characterize keloid fibroblasts; identifying such genetic disequilibrium may shed light on the molecular signaling events responsible for keloid formation. In this study, we performed gene expression analysis of fibroblasts isolated from keloid lesions from three individuals in comparison with the fibroblasts isolated from normal skin using the Affymetrix U133a chip (22,284 genes and expression sequence tags). We found through J5 test score expression analysis that among 22,284 genes, there were 43 genes that were overexpressed and five genes were underexpressed in keloid fibroblasts when compared with dermal fibroblasts from persons without keloids. The overexpression of three genes not previously reported as being up-regulated in keloids (annexin A2, Transgelin, and RPS18) was confirmed by real-time polymerase chain reaction. Certain overexpressed genes were similar to previous biochemical observations on the protein levels of these overexpressed genes during keloid formation. We also report for the first time that a few tumor-related genes are overexpressed in keloid fibroblasts.     

瘢痕疙瘩家系Fas基因死亡域突变的实验研究

目的 探讨瘢痕疙瘩家系标本中Fas基因（外显子7～9）有无突变以及Fas基因突变在瘢痕疙瘩形成中的意义。方法 实验标本来自南方医科大学南方医院整形外科2005年收集的A、B两个瘢痕疙瘩家系。采用PCR及基因测序技术，分别以A家系2例男性患者的瘢痕疙瘩组织为研究对象，以其周围正常皮肤及外周静脉血作为自身对照，其配偶的外周静脉血作为正常对照，并以B家系2例患者（母亲与儿子）的外周静脉血作为不同家系之间的对照，共检测10份标本中Fas基因外显子7～9基因序列。结果 10份瘢痕疙瘩家系标本Fas基因的7、8外显子均未发生突变，2例瘢痕疙瘩组织标本均在外显子9编码区的11bp和53bp两个位点上存在单个碱基基因突变或多态性改变。结论 瘢痕疙瘩Fas基因死亡域外显子9区段的基因结构异常，极有可能与Fas蛋白的功能改变有关，从而导致局部瘢痕疙瘩形成。     

瘢痕疙瘩相关基因的生物信息学分析

目的比较瘢痕疙瘩与正常皮肤的基因表达差异,从分子水平探讨瘢痕疙瘩的发病机制,为临床治疗提供新思路。方法用PubMed数据库文献检索瘢痕疙瘩与正常皮肤的差异表达基因,对与瘢痕疙瘩相关的基因进行蛋白-蛋白相互作用网络、生物学通路、基因本体(gene ontology,GO)和功能注释聚类的生物信息学分析。结果获得差异表达基因谱8个和文献922篇,筛选瘢痕疙瘩相关基因94个(71个上调,23个下调)。86个基因构成蛋白-蛋白相互作用网络,TGFB1、FN1、COL1A1、MMP9、VEGFA、TP53、IL6和MMP2为核心蛋白。瘢痕疙瘩相关基因参与信号转导、肿瘤形成等生物学通路,细胞凋亡、细胞运动等生物过程,形成细胞膜结构和细胞外基质、胶原等组分。结论 TGFB1、FN1、COL1A1、MMP9、VEGFA、TP53、IL6和MMP2等关键基因,TGF-β信号转导、细胞增殖和凋亡、肿瘤形成相关通路在瘢痕疙瘩的发生发展中可能起重要作用。