本文選題：小梁切除　切入點(diǎn)：抗瘢痕　出處：《第三軍醫(yī)大學(xué)》2010年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】： 青光眼濾過(guò)術(shù)是青光眼的一種常規(guī)手術(shù)方式,手術(shù)的成功率與術(shù)區(qū)的瘢痕化密切相關(guān)。為了提高手術(shù)成功率,術(shù)中常應(yīng)用抗代謝藥物來(lái)預(yù)防術(shù)后傷口瘢痕化。雖然絲裂霉素-C(MMC)、5-氟尿嘧啶(5-Fu)等抗代謝藥物術(shù)中應(yīng)用提高了手術(shù)成功率,但術(shù)后發(fā)生的持續(xù)性低眼壓、濾過(guò)泡滲漏、眼內(nèi)炎等并發(fā)癥也隨之增多。為了減少術(shù)后濾過(guò)道瘢痕形成,各種抗瘢痕化學(xué)、生物制劑及物理粘附屏障已經(jīng)運(yùn)用于動(dòng)物濾過(guò)術(shù)模型中,并取得了不錯(cuò)的治療效果,但是由于各種原因,都還沒(méi)有應(yīng)用于臨床。 ski基因是一種原癌基因,在體內(nèi)作為轉(zhuǎn)錄的輔活化子或輔阻遏子調(diào)節(jié)多個(gè)重要轉(zhuǎn)錄因子的表達(dá),對(duì)創(chuàng)口組織再生起著重要的作用。大坪醫(yī)院野戰(zhàn)外科研究所第七研究室前期研究發(fā)現(xiàn),C-ski能促進(jìn)體外培養(yǎng)的大鼠皮膚成纖維細(xì)胞增殖和降低膠原分泌,對(duì)兔耳創(chuàng)口也有促進(jìn)愈合,降低瘢痕的作用,因此推測(cè)C-ski基因可能對(duì)促進(jìn)創(chuàng)傷愈合,降低瘢痕形成有重要意義,為此,我們將其用于兔眼小梁切除術(shù)后的抗瘢痕研究,觀察其療效,為以后的臨床應(yīng)用奠定基礎(chǔ)。 本實(shí)驗(yàn)主要進(jìn)行了以下三個(gè)方面的研究:1.對(duì)C-ski基因重組pcDNA3.0質(zhì)粒及pcDNA3.0空載質(zhì)粒進(jìn)行擴(kuò)增、提純并加以鑒定;2.不同劑量的C-ski基因重組pcDNA3.0質(zhì)粒及PBS對(duì)兔眼小梁切除后濾過(guò)道瘢痕形成的影響;3. 500ug劑量C-ski基因重組pcDNA3.0質(zhì)粒及絲裂霉素、PBS、空載質(zhì)粒對(duì)兔眼小梁切除后濾過(guò)道瘢痕形成的影響。 主要結(jié)果和結(jié)論如下: 1.通過(guò)對(duì)C-ski基因重組pcDNA3.0質(zhì)粒及pcDNA3.0空載質(zhì)粒進(jìn)行擴(kuò)增、純化和鑒定,結(jié)果證實(shí):采用堿裂解法提取的C-ski基因重組pcDNA3.0質(zhì)粒及pcDNA3.0空載質(zhì)粒的質(zhì)和量,能夠達(dá)到我們后續(xù)動(dòng)物實(shí)驗(yàn)的要求。 2.不同劑量的C-ski基因重組pcDNA3.0質(zhì)粒能降低眼壓及延長(zhǎng)濾過(guò)泡存留時(shí)間,并且能抑制濾過(guò)道Ⅰ型膠原生成,劑量增加,效果增強(qiáng),而對(duì)濾過(guò)道成纖維細(xì)胞及肌成纖維細(xì)胞生成及轉(zhuǎn)化無(wú)明顯影響。 3. 500ug劑量的C-ski基因重組質(zhì)粒對(duì)兔眼小梁切除術(shù)后進(jìn)行抗瘢痕治療時(shí),未引起炎癥反應(yīng),能明顯減少Ⅰ型膠原生成量,抑制濾過(guò)道瘢痕的形成,延長(zhǎng)濾過(guò)術(shù)后低眼壓的時(shí)間,而無(wú)絲裂霉素(MMC)副作用。
[Abstract]:Glaucoma filtration is a routine operation for glaucoma. The success rate of operation is closely related to scar change in operation area. Antimetabolic drugs are often used to prevent scar formation after operation. Although the use of anti-metabolic drugs such as mitomycin CMMC and 5-fluorouracil (5-Fu) improves the success rate of the operation, the persistent intraocular pressure and bleb leakage occur after the operation. In order to reduce the formation of post-operative filtration scar, various anti-scar chemical, biological agents and physical adhesion barrier have been used in animal model of filtration surgery, and have achieved good therapeutic effect. However, due to various reasons, has not been applied to clinical. Ski gene is a proto-oncogene. As a coactivator or repressor of transcription in vivo, it regulates the expression of many important transcription factors. It was found that C-ski could promote the proliferation and decrease of collagen secretion of rat skin fibroblasts cultured in vitro, and promote the healing of rabbit ear wounds. Therefore, we speculate that C-ski gene may play an important role in promoting wound healing and reducing scar formation. Therefore, we used it to study the anti-scar effect after trabeculectomy in rabbits. To lay a foundation for clinical application in the future. This experiment mainly carried out the following three aspects of research: 1.Amplification of C-ski gene recombinant pcDNA3.0 plasmid and pcDNA3.0 empty plasmid, Effect of different doses of C-ski Gene Recombinant pcDNA3.0 plasmid and PBS on the formation of Trabeculectomy scar in Rabbit eyes after trabeculectomy; 3500ug / g C-ski Gene Recombinant pcDNA3.0 plasmid and mitomycin PBS; No-loaded plasmid for trabeculectomy in Rabbit eyes. The effect of retrofiltration on scar formation in the passageway. The main findings and conclusions are as follows:. 1. The C-ski gene recombinant pcDNA3.0 plasmid and pcDNA3.0 empty plasmid were amplified, purified and identified. The results showed that the quantity and quality of C-ski gene recombinant pcDNA3.0 plasmid and pcDNA3.0 empty plasmid were obtained by alkaline lysis method. To meet the requirements of our subsequent animal experiments. 2. Different doses of C-ski gene recombinant pcDNA3.0 plasmid can reduce intraocular pressure and prolong the retention time of bleb, and can inhibit the formation of collagen type I in the filtering passage, and the dose is increased and the effect is enhanced. However, there was no significant effect on the formation and transformation of fibroblasts and myofibroblasts. After trabeculectomy, 3500ug dose of C-ski gene recombinant plasmid did not cause inflammatory reaction, significantly reduced collagen I production, inhibited the formation of filtering scar, and prolonged the time of low intraocular pressure after filtering. There were no side effects of MMC.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R779.6

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