系统性硬皮病患者皮肤克隆成纤维细胞胶原代谢的异质性

目的在克隆水平上研究系统性硬皮病(SSc)和正常皮肤成纤维细胞(FB)胶原代谢异质性的特点。方法以皮肤克隆FB作为研究对象,采用实时荧光定量RT-PCR法检测其Ⅰ,Ⅲ型前胶原和基质金属蛋白酶(MMP-1,MMP-3)mRNA水平;采用酶联免疫吸附法检测其细胞培养上清中Ⅰ型胶原蛋白的表达,筛选出具有不同胶原合成能力的异质性克隆,分析其胶原代谢的特点。结果 SSc和正常皮肤克隆FB的胶原代谢均具有异质性,SSc患者克隆FBⅠ,Ⅲ型前胶原mRNA水平的异质性较正常人大(F=20.540,P=0.000;F=6.822,P=0.012),SSc患者Ⅰ型前胶原mRNA高表达克隆的比例高于正常人(48.0%vs14.3%);SSc和正常皮肤克隆FB细胞培养上清中Ⅰ型胶原蛋白水平与其mRNA水平均存在正相关关系(r=0.873,P=0.000;r=0.538,P=0.039)。结论 SSc和正常皮肤克隆FB的胶原代谢均具有异质性。     

活血化瘀中药对系统性硬皮病患者皮肤成纤维细胞胶原合成的影响

目的研究活血化瘀中药对系统性硬皮病(SSc)患者皮肤成纤维细胞胶原合成的影响.方法以组织块培养法建立3个SSc患者皮肤成纤维细胞株.将活血化瘀中药加入成纤维细胞培养液,其浓度分别为5g/L、2.5g/L和1.25g/L,于加药后第3、6 d,以分光光度法测定细胞培养上清液中羟脯氨酸浓度并折算成胶原浓度.结果积雪草和丹参在5g/L、2.5g/L及1.25g/L浓度下于作用第3 d和第6 d时能显著抑制SSc患者皮肤成纤维细胞的胶原合成(P＜0.05～0.01),红花在5 g/L、2.5g/L及1.25 g/L浓度下于作用第3 d和第6 d时能显著促进SSc患者皮肤成纤维细胞的胶原合成(P＜0.05).此外,当归在5g/L、茜草在2.5g/L浓度下于作用第6 d时也能显著抑制SSc患者皮肤成纤维细胞合成胶原(P＜0.05).结论不同的活血化瘀中药对皮肤成纤维细胞胶原合成的作用不一,该结果可为临床制定治疗SSc及其他相关疾病的中药方剂提供参考.     

丹参对系统性硬皮病患者皮肤成纤维细胞增殖和胶原合成的影响

目的　探讨丹参治疗系统性硬皮病的药理机制。方法　用四甲基偶氮唑盐 (MTT)方法和光度检测法测定了丹参对成纤维细胞增殖和培养基中胶原含量的影响。结果　丹参水溶性提取物丹参注射液 (10 m g/m l、7.5 m g/ml、 5 .0 m g/m l、2 .5 m g/ml)及其活性单体丹参素 (6 0 μg/m l、 45 μg/ml、 30 μg/m l、15 μg/m l)和原儿茶醛 (40 μg/m l、2 0 μg/m l、10 μg/m l、5 μg/m l) ,以及丹参脂溶性总提取物 (1mg/m l、0 .5 m g/m l、0 .2 5 m g/m l)及其活性单体丹参酮 A (4μg/ml,2μg/m l) ,不仅对患者皮肤成纤维细胞的增殖具有显著的抑制作用 ,还能显著降低该细胞培养基中可溶性胶原的含量 (P<0 .0 5～ 0 .0 1)。上述抑制作用随药物浓度的增高或作用时间的延长而增强。采用流式细胞仪对该细胞周期的检测显示 ,丹参素组 G0 /G1 期细胞 ,原儿茶醛组和丹参酮 A组 G2 M期细胞的比例较正常人对照组细胞显著增高 (P<0 .0 1)。结论　丹参对体外培养的系统性硬皮病患者的皮肤成纤维细胞的增殖和胶原合成具有显著的抑制作用 ,可能是其治疗系统性硬皮病的药理机理之一。     

硬皮病皮肤硬化的中西医治疗进展

硬皮病是结缔组织病中较为常见的疾病,大量研究表明它的发病与组织纤维化、自身免疫反应和血管功能异常三个基本机制有关.现有的硬皮病的治疗措施主要是基于本病发病涉及的三个基本机制进行治疗.     

系统性硬皮病患者缺氧与皮肤硬化关系的初探

目的 探讨缺氧对系统性硬皮病 (SSc)患者皮肤硬化的影响。方法 测定 SSc患者血氧分压和皮肤硬度积分,对二者进行直线相关分析,并测定低氧 (2.5％氧浓度 )对体外培养的 SSc患者皮肤成纤维细胞增殖的影响。结果 30例 SSc患者血氧分压为 6.38～ 12.06 kPa,平均 (9.53± 1.93)kPa; 20例正常对照组为 (10.67～ 13.33)kPa,平均为 (12.01± 0.81)kPa。前者比后者显著降低 (t=5.276,P     

不同性别系统性硬皮病患者临床表现分析

系统性硬皮病(systemic sclerosis,SSc)是一种以皮肤及各系统胶原纤维进行性硬化为特征的结缔组织疾病,好发于女性,男性较少.为了解男性与女性SSc患者在临床表现与实验室检查方面的异同.     

系统性硬皮病患者血清肿瘤坏死因子的测定

系统性硬皮病患者血清肿瘤坏死因子的测定蔡茂庆①王守义②钮斌①孙家英①屠文震①程培俊③张维萍③沈建人③肿瘤坏死因子（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ，ＴＮＦ）是８０年代发现的由巨噬细胞和单核细胞分泌的一种重要的细胞因子。具有多种生物学效应，主要是...     

低氧对系统性硬化病患者皮肤成纤维细胞胶原蛋白合成的影响

目的 研究低氧对系统性硬化病(SSc)患者和正常人皮肤成纤维细胞胶原蛋白合成及胶原基因表达的影响。方法 分别以5例SSc患者和5例正常人皮肤成纤维细胞系作为实验对象。将成纤维细胞分别放入20%正常氧和2%低氧分压环境中培养,用分光光度仪测定细胞培养基上清液中羟脯氨酸含量,并折算成胶原蛋白含量。用Trizol提取细胞总RNA,用RT-PCR方法测定其中Ⅰ型和Ⅲ型前胶原mRNA的表达。结果 培养至第6天,低氧分压条件下,SSc患者皮肤成纤维细胞培养基中胶原蛋白含量比正常氧分压条件下显著增加(t=3.97,P=0.0041);正常人皮肤成纤维细胞培养基中胶原蛋白含量此时也显著增加(t=2.63,P=0.0302)。培养至第3天,低氧分压条件下,SSc成纤维细胞Ⅰ型和Ⅲ型前胶原mRNA表达量比正常氧分压条件下均有显著增加(Ⅰ型:t=5.81,P=0.0004;Ⅲ型:t=6.44,P=0.0002);正常人皮肤成纤维细胞此时的Ⅰ型和Ⅲ型前胶原mRNA表达量与SSc患者相似,低氧分压条件下比正常氧分压条件下均有显著增加(Ⅰ型:t=4.40,P=0.0023;Ⅲ型:t=2.24,P=0.0453)。结论 低氧状态下,SSc患者和正常人皮肤成纤维细胞胶原蛋白合成均有增加,Ⅰ型和Ⅲ型前胶原mRNA表达也均增加.低氧可能是皮肤硬化的一个相关因素。     

Inhibition of collagen expression by azelastine hydrochloride in cultured skin fibroblasts from normal individuals and scleroderma patients

The effects of azelastine hydrochloride on cell proliferation and collagen synthesis in cultured human skin fibroblasts were studied. Azelastine inhibited cell proliferation during proliferating cell phases. Azelastine was found to inhibit collagen synthesis without altering cell proliferation during quiescent phases. It did not alter the ratio of type I to III collagen synthesis. Northern blot analysis of collagen chain mRNAs revealed that the levels of α₁ (I), α₁ (III) and α₁ (VI) mRNAs were reduced by azelastine treatment, whereas the level of α₂ (VI), α₃ (VI) mRNAs were not significantly changed. These results suggest that azelastine modulates collagen synthesis at a pretranslational level. Azelastine inhibited collagen synthesis in fibroblasts from scleroderma patients to the same extent as in normal skin fibroblasts. This drug may be useful in the treatment of fibrotic diseases.     

局限性硬皮病发病机制的研究进展

局限性硬皮病是一种以局部皮肤及皮下组织纤维化为特征的疾病.病因不明,发病机制可能涉及小血管的损伤、T细胞的活化以及结缔组织生成增加等多个过程.T细胞活化释放各种细胞因子,趋化因子,主要包括肿瘤坏死因子α、转化生长因子β、可溶性白细胞介素受体2和可溶性白细胞介素受体6等.血管内皮细胞的损伤也可介导纤维化相关的细胞因子释放.局限性硬皮病和系统性硬皮病在发病机制上有一定的区别和联系,概述局限性硬皮病发病机制的最新进展.     

Aminoterminal extension peptides from type I procollagen normalize excessive collagen synthesis of scleroderma fibroblasts

Summary Fibroblasts derived from a skin biopsy of a patient with scleroderma in the sclerotic stage were shown to have a higher rate of DNA synthesis, and to synthesize more collagen than fibroblasts from a healthy control. The addition of procollagen peptides to the culture medium of scleroderma fibroblasts almost normalized the collagen synthesis. This observation indicates that the mechanism for the regulation of collagen synthesis by feed back inhibition of prollagen peptides is functioning in this disease. It is suggested that the level of biologically active procollagen peptides is lowered.

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Abbreviations BAPN -aminopropriontrile - CM carboxymethyl - DNA desoxyribonucleic acid - NCP non-collagenous proteins - MEM minimum essential medium     

体外培养硬皮病成纤维细胞的超微结构观察

为探讨硬皮病纤维化硬化形成的机制，应用透射电镜对培养的硬皮病皮损和正常皮肤中的成纤维细胞进行了超微结构观察。结果示硬皮病成纤维细胞中有丰富的，高度发达的粗面内质网，且多个区域有高尔基复合体及其形成的分泌泡存在。正常皮肤中的成纤维细胞内粗面内质网少，未见高尔基复合体的分泌泡。提示硬皮病成纤维细胞的蛋白合成及分泌功能活跃，因此胶原合成增高。     

Collagen specific amino acids in skin in localized scleroderma

Hydroxyproline, hydroxylysine and proline were determined on skin from 18 patients with localized scleroderma (10 with localized morphoea plaque and 8 with generalized morphoea). Three skin biopsies (4mm punch) were obtained from each patient: One from the center of a sclerotic plaque, one from the perilesional area, and one (control) from unaffected skin of the same region. Clinically, the sclerosis was more pronounced (p less than 0.01) in localized morphoea plaque as compared to generalized morphoea. Patients with localized morphoea plaque had an increased concentration of hydroxylysine (p less than 0.01) and an increased ratio of hydroxylysine to hydroxyproline (p less than 0.01) in the plaques. Hydroxylysine concentration was not changed in patients with generalized morphoea. In the entire material, increased hydroxylysine concentration were related to shorter age of the plaques (p less than 0.05) and to advanced degree of sclerosis (p less than 0.05). The hydroxylysine and hydroxyproline content per mm2 skin surface, and the weight of the dried defatted biopsy cores were increased in sclerotic plaques (p less than 0.01) in localized as well as generalized morphoea. There were no changes in the hydroxyproline and proline concentrations in any of the groups. Specimens from perilesional area showed intermediate changes. The results were compared with selected cases of lichen sclerosis et atrophicus and atrophic skin diseases. The increase in hydroxylysine concentration and ratio to hydroxyproline indicate that patients with localized morphoea plaque contain an increased proportion of newly synthesized collagen in the fibrotic plaque.     

Clinical evaluation of scleroderma spectrum disorders using a points system

We have established a new diagnostic method using a points system to evaluate patients with early scleroderma and those with scleroderma spectrum disorders (SSD). To examine the clinical usefulness of this method, it was applied to a total of 215 cases including 97 patients with scleroderma, 32 with SSD, 28 with presumed primary Raynaud's phenomenon (RP) and 58 with other connective tissue disorders (CTD). A total score was obtained for each patient as the sum of the following five factors: (1) extent of skin sclerosis (maximum, 10 points); (2) pulmonary changes (maximum, 4 points); (3) antinuclear antibodies (maximum, 5 points); (4) pattern of Raynaud's phenomenon (maximum, 3 points); and (5) nailfold bleeding (maximum, 2 points). Of the 97 scleroderma patients, 86 (89%) had 9 or more points, and of the 32 SSD patients, 28 (88%) had 5 to 8 points. In contrast, all patients with presumed primary RP and 54 of 58 (93%) patients with other CTD had 0 to 4 points. These data suggest that this diagnostic method is very useful not only for clinical evaluation of SSD, but also for the differentiation of scleroderma and SSD from other CTD and primary RP.     

系统性硬皮病105例临床分析

对１０５例系统性硬皮病患的一般情况、临床表现了详细分析。男女患者之比为１：４，发病年龄１５岁－７２岁，２１－５０岁患者占大多数，所有患者均有不同程度的皮肤改变，皮受累范围大小不一。９９％，患者嗓手皮肤受累，８１％患者有雷诺现象，６７例患者内脏器官受累。还对其实验室检查，伴发病、病因、诊断和治疗作了分析和讨论。     

硬皮病动物模型的应用进展

硬皮病是一种自身免疫性结缔组织疾病，严重影响患者的生活质量。该病的病因和发病机制尚未完全明了，临床上也无理想的治疗方法，在进行基础研究时患者依从性相对较差。因此建立硬皮病的动物模型，对于深入研究本病的发病机制和评价治疗方法有重要意义。综述近十年来利用不同品系的鼠科动物建立的各种硬皮病模型，以及利用这些模型从不同的角度阐明硬皮病可能的发病机制．并在新药筛选和新疗法评价方面取得的一些进展。