姜黄素抗增生性瘢痕作用及其机制的实验研究
发布时间：2018-03-30 00:36
本文选题：姜黄素 切入点：增生性瘢痕 出处：《第二军医大学》2015年硕士论文
【摘要】：增生性瘢痕常见于外科手术后的恢复阶段,其中以整形外科最为多见,常在机体组织皮肤的完整性遭到破坏后发生,现对其发生的具体原因尚不了解,但大量临床研究结果显示,组织受损后的恢复期间,机体会通过一定的途径刺激受损组织的细胞外基质分泌相应的细胞因子,从而促进受损部位的细胞进行大量的生长繁殖。根据增生性瘢痕多个部位、多个环节相互作用进行调控的机制,现从中药中提取出有效活性成分,证明在治疗增生性瘢痕方面有一定的应用价值。姜黄属于我国的一类常用中药,其药性温和但味辛、苦,其功效主要表现为通气、活血、疏经。姜黄素是从姜黄中提取的一种多酚类物质,其主要作用是消除炎症、抵抗自由基、抑制肿瘤的发生发展等。本实验通过对姜黄素治疗增生性瘢痕的作用机制研究,以期为姜黄素临床治疗增生性瘢痕提供理论依据。第一部分姜黄素减轻兔耳增生性瘢痕作用的研究目的:研究在兔耳增生性瘢痕中姜黄素发挥的作用。方法:在兔耳的腹侧面模拟增生性瘢痕,实验分为正常皮肤组(A组)、瘢痕模型组(B组)、康瑞保#174;阳性药对照组(C组)、小剂量姜黄素组(D组)、中等剂量姜黄素组(E组)、大剂量姜黄素组(F组),每组16个增生性瘢痕点(A组相同位置16个正常皮肤点),术后28 d开始涂抹给药治疗,0.1 g,1次/d,连续给药28 d后收取各组标本。HE染色、Masson染色观察,测量瘢痕增生的变化情况、成纤维细胞和胶原纤维分裂繁殖的情况。结果:姜黄素可以明显抑制瘢痕的生成与发展,使其体积缩小、质地柔软,接近正常组织,生长较快的成纤维细胞在数量方面明显下降,胶原纤维在数量方面也明显下降、分布逐渐规则、致密度逐渐下降;瘢痕增生指数随姜黄素浓度的增加显著下降,差异有统计学意义(P0.05);成纤维细胞密度、胶原纤维面密度降低,差异有统计学意义(P0.05)。结论:姜黄素在兔耳增生性瘢痕方面具有显著地治疗作用,通过进一步的药理研究、开发新剂型,姜黄素可能发展成为治疗与预防增生性瘢痕的一线药物。第二部分姜黄素对兔耳增生性瘢痕胶原合成与降解的影响目的:研究姜黄素对兔耳增生性瘢痕CollagenⅠ、CollagenⅢ、MMP-1、TIMP-1mRNA表达量和TGF-β1蛋白表达量的影响。方法:在兔耳的腹侧面模拟增生性瘢痕,实验分为正常皮肤组(A组)、瘢痕模型组(B组)、康瑞保#174;阳性药对照组(C组)、小剂量姜黄素组(D组)、中等剂量姜黄素组(E组)、大剂量姜黄素组(F组),每组16个增生性瘢痕点(A组是相同位置16个正常皮肤点),术后28 d开始涂抹给药治疗,0.1 g,1次/d,连续给药28 d后收取各组标本。Real-time PCR检测CollagenⅠ、CollagenⅢ、MMP-1、TIMP-1 mRNA表达量,Western Blot检测TGF-β1蛋白表达量。结果:姜黄素组CollagenⅠmRAN表达量呈剂量依赖性下降;姜黄素组CollagenⅢmRAN表达量比模型组显著降低;中等、大剂量姜黄素组MMP-1 mRAN表达量比模型组进一步增高;姜黄素组TIMP-1 mRAN表达量比模型组显著降低;姜黄素组TGF-β1蛋白表达量呈剂量依赖性下降。结论:姜黄素能够降低CollagenⅠ、CollagenⅢmRNA表达,增加MMP-1 mRNA表达、降低TIMP-1 mRNA表达,同时降低TGF-β1蛋白表达,这可能是姜黄素治疗兔耳增生性瘢痕的机制之一。第三部分姜黄素诱导人增生性瘢痕成纤维细胞凋亡的机制研究目的:研究姜黄素诱导增生性瘢痕成纤维细胞凋亡及其分子机制。方法:用10、20、40μmol/L姜黄素(对照组0μmol/L)处理增生性瘢痕成纤维细胞24 h,MTT法检测姜黄素对细胞增殖的影响,Annexin V-FITC/PI双标法检测细胞凋亡率,分光光度法检测Caspase-9、Caspase-3活性,Western Blot法检测Bcl-2、Bax蛋白表达。结果:MTT结果显示姜黄素可以显著控制瘢痕处成纤维细胞的生长数量,并且姜黄素的这种作用与其浓度呈正相关(P0.05);Annexin V-FITC/PI双标法结果显示成纤维细胞凋亡率随姜黄素浓度增加而上升(P0.05);经姜黄素作用,Caspase-9、Caspase-3的活性随姜黄素浓度增加而升高(P0.05);Western Blot结果显示Bcl-2蛋白表达下降,Bax蛋白表达升高,Bax/Bcl-2比值增大,差异有统计学意义(P0.05)。结论:姜黄素对增生性瘢痕成纤维细胞有抑制增殖和诱导凋亡的作用,其机制与下调Bcl-2蛋白表达、上调Bax蛋白表达,升高Caspase-9、Caspase-3活性有关。
[Abstract]:The common hypertrophic scar after surgery in the recovery phase, in which plastic surgery is the most common, often occur in the integrity of the body the skin is damaged, the specific reason of its occurrence is not known, but a large number of clinical studies showed that during the recovery of damaged tissue, body through certain the way of extracellular matrix stimulation of damaged tissue cytokine secretion accordingly, so as to promote the damaged parts of the cell growth and reproduction. According to a large number of hypertrophic scar parts. The mechanism of multiple aspects of interaction control, the active ingredient extracted from Chinese herbal medicine, that has a certain application value in the treatment of hyperplasia scar. A kind of traditional Chinese medicine Jiang Huang belongs to China, but its mild spicy, bitter, its function mainly for ventilation, blood, dredging. Curcumin is extracted from a yellow ginger. Polyphenols, its main role is to eliminate inflammation, resist free radical and inhibit tumor development. Study on Mechanism of curcumin in the treatment of hypertrophic scar by this experiment, in order to provide theoretical basis for clinical treatment of hypertrophic scar. Curcumin on the first part of Jiang Huang - reducing effect of hypertrophic scar in rabbit ears: Study on the play the effect of curcumin on hypertrophic scar of rabbit ear in vitro. Methods: Simulation of hypertrophic scar in rabbit ear ventral side, the experiment was divided into normal skin group (A group), scar model group (B group), positive drug Contractubex #174; the control group (C group), low dose curcumin group (D group) that middle dose curcumin group (E group), high dose of curcumin group (F group), each group of 16 points of hypertrophic scar (group A, 16 normal skin at the same position), 28 d after the operation began to apply to the treatment, 0.1 g, 1 /d, 28 d after continuous administration of collection of specimens.HE dye Color, Masson staining, changes of hypertrophic scar measurement, fibroblasts and collagen fibers proliferation situation. Results: Curcumin could significantly inhibit the formation and development of scar, to reduce volume, soft texture, close to the normal tissue, growth of fibroblasts decreased significantly in terms of quantity, the collagen fibers were decreased in terms of quantity, distribution gradually rules, density gradually decreased; increase the hypertrophic index with the concentration of curcumin significantly decreased, the difference was statistically significant (P0.05); fibroblast density, collagen fiber density decreased, the difference was statistically significant (P0.05). Conclusion: curcumin has a significant role in the treatment of hypertrophic scar of rabbit ear hyperplasia, through further pharmacological studies, the development of new form, curcumin may become a first-line drug for the treatment and prevention of hypertrophic scar. The second part of Curcumin 瀵瑰厰鑰冲鐢熸，
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