本文選題：姜黃素　切入點：增生性瘢痕　出處：《第二軍醫(yī)大學(xué)》2015年碩士論文

【摘要】：增生性瘢痕常見于外科手術(shù)后的恢復(fù)階段,其中以整形外科最為多見,常在機體組織皮膚的完整性遭到破壞后發(fā)生,現(xiàn)對其發(fā)生的具體原因尚不了解,但大量臨床研究結(jié)果顯示,組織受損后的恢復(fù)期間,機體會通過一定的途徑刺激受損組織的細(xì)胞外基質(zhì)分泌相應(yīng)的細(xì)胞因子,從而促進受損部位的細(xì)胞進行大量的生長繁殖。根據(jù)增生性瘢痕多個部位、多個環(huán)節(jié)相互作用進行調(diào)控的機制,現(xiàn)從中藥中提取出有效活性成分,證明在治療增生性瘢痕方面有一定的應(yīng)用價值。姜黃屬于我國的一類常用中藥,其藥性溫和但味辛、苦,其功效主要表現(xiàn)為通氣、活血、疏經(jīng)。姜黃素是從姜黃中提取的一種多酚類物質(zhì),其主要作用是消除炎癥、抵抗自由基、抑制腫瘤的發(fā)生發(fā)展等。本實驗通過對姜黃素治療增生性瘢痕的作用機制研究,以期為姜黃素臨床治療增生性瘢痕提供理論依據(jù)。第一部分姜黃素減輕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黃素發(fā)揮的作用。方法:在兔耳的腹側(cè)面模擬增生性瘢痕,實驗分為正常皮膚組(A組)、瘢痕模型組(B組)、康瑞保#174;陽性藥對照組(C組)、小劑量姜黃素組(D組)、中等劑量姜黃素組(E組)、大劑量姜黃素組(F組),每組16個增生性瘢痕點(A組相同位置16個正常皮膚點),術(shù)后28 d開始涂抹給藥治療,0.1 g,1次/d,連續(xù)給藥28 d后收取各組標(biāo)本。HE染色、Masson染色觀察,測量瘢痕增生的變化情況、成纖維細(xì)胞和膠原纖維分裂繁殖的情況。結(jié)果:姜黃素可以明顯抑制瘢痕的生成與發(fā)展,使其體積縮小、質(zhì)地柔軟,接近正常組織,生長較快的成纖維細(xì)胞在數(shù)量方面明顯下降,膠原纖維在數(shù)量方面也明顯下降、分布逐漸規(guī)則、致密度逐漸下降;瘢痕增生指數(shù)隨姜黃素濃度的增加顯著下降,差異有統(tǒng)計學(xué)意義(P0.05);成纖維細(xì)胞密度、膠原纖維面密度降低,差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:姜黃素在兔耳增生性瘢痕方面具有顯著地治療作用,通過進一步的藥理研究、開發(fā)新劑型,姜黃素可能發(fā)展成為治療與預(yù)防增生性瘢痕的一線藥物。第二部分姜黃素對兔耳增生性瘢痕膠原合成與降解的影響目的:研究姜黃素對兔耳增生性瘢痕CollagenⅠ、CollagenⅢ、MMP-1、TIMP-1mRNA表達量和TGF-β1蛋白表達量的影響。方法:在兔耳的腹側(cè)面模擬增生性瘢痕,實驗分為正常皮膚組(A組)、瘢痕模型組(B組)、康瑞保#174;陽性藥對照組(C組)、小劑量姜黃素組(D組)、中等劑量姜黃素組(E組)、大劑量姜黃素組(F組),每組16個增生性瘢痕點(A組是相同位置16個正常皮膚點),術(shù)后28 d開始涂抹給藥治療,0.1 g,1次/d,連續(xù)給藥28 d后收取各組標(biāo)本。Real-time PCR檢測CollagenⅠ、CollagenⅢ、MMP-1、TIMP-1 mRNA表達量,Western Blot檢測TGF-β1蛋白表達量。結(jié)果:姜黃素組CollagenⅠmRAN表達量呈劑量依賴性下降;姜黃素組CollagenⅢmRAN表達量比模型組顯著降低;中等、大劑量姜黃素組MMP-1 mRAN表達量比模型組進一步增高;姜黃素組TIMP-1 mRAN表達量比模型組顯著降低;姜黃素組TGF-β1蛋白表達量呈劑量依賴性下降。結(jié)論:姜黃素能夠降低CollagenⅠ、CollagenⅢmRNA表達,增加MMP-1 mRNA表達、降低TIMP-1 mRNA表達,同時降低TGF-β1蛋白表達,這可能是姜黃素治療兔耳增生性瘢痕的機制之一。第三部分姜黃素誘導(dǎo)人增生性瘢痕成纖維細(xì)胞凋亡的機制研究目的:研究姜黃素誘導(dǎo)增生性瘢痕成纖維細(xì)胞凋亡及其分子機制。方法:用10、20、40μmol/L姜黃素(對照組0μmol/L)處理增生性瘢痕成纖維細(xì)胞24 h,MTT法檢測姜黃素對細(xì)胞增殖的影響,Annexin V-FITC/PI雙標(biāo)法檢測細(xì)胞凋亡率,分光光度法檢測Caspase-9、Caspase-3活性,Western Blot法檢測Bcl-2、Bax蛋白表達。結(jié)果:MTT結(jié)果顯示姜黃素可以顯著控制瘢痕處成纖維細(xì)胞的生長數(shù)量,并且姜黃素的這種作用與其濃度呈正相關(guān)(P0.05);Annexin V-FITC/PI雙標(biāo)法結(jié)果顯示成纖維細(xì)胞凋亡率隨姜黃素濃度增加而上升(P0.05);經(jīng)姜黃素作用,Caspase-9、Caspase-3的活性隨姜黃素濃度增加而升高(P0.05);Western Blot結(jié)果顯示Bcl-2蛋白表達下降,Bax蛋白表達升高,Bax/Bcl-2比值增大,差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:姜黃素對增生性瘢痕成纖維細(xì)胞有抑制增殖和誘導(dǎo)凋亡的作用,其機制與下調(diào)Bcl-2蛋白表達、上調(diào)Bax蛋白表達,升高Caspase-9、Caspase-3活性有關(guān)。
[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 瀵瑰厰鑰沖鐢熸，

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