【摘要】：目的：研究基質(zhì)金屬蛋白酶(MMPs)和基質(zhì)金屬蛋白酶組織抑制因子(TIMPs)在不同發(fā)育時(shí)期的胎兒皮膚和不同形成時(shí)期的增生性瘢痕中基因表達(dá)和蛋白含量、分布的變化，探索無瘢痕愈合發(fā)生機(jī)制以及增生性瘢痕發(fā)生和成熟的部分機(jī)制。 方法：①胎兒皮膚因創(chuàng)傷等意外因素終止妊娠的胎兒背部全層皮膚，共18例，胎齡(EGA)13～33周。標(biāo)本按胎齡分為3組，即早期(13～19周)，中期(21～25周)和晚期(27～33周)，每組6例。②增生性瘢痕組織取自進(jìn)行整形手術(shù)患者的增生性瘢痕16例，平均年齡為26歲，傷口愈合的時(shí)間為4月～11年。瘢痕增生時(shí)間1年以內(nèi)8例，1～2年4例，2年以上4例。正常對(duì)照組皮膚8例，取自病人供皮區(qū)的全層皮膚。標(biāo)本經(jīng)無菌取材后，一部分立即液氮凍存，用于進(jìn)行逆轉(zhuǎn)錄-多聚酶鏈反應(yīng)(RT-PCR)和Western blot實(shí)驗(yàn)，檢測(cè)MMP-1，MMP-2，MMP-3，MMP-7，MMP-9，MMP-14，TIMP-1，TIMP-2和TIMP-3基因表達(dá)和蛋白含量的變化；另一部分標(biāo)本經(jīng)10％中性甲醛液固定，脫水，石蠟包埋后切片，用于免疫組化實(shí)驗(yàn)，鏡下觀察。 結(jié)果：1．在早期妊娠胎兒皮膚中MMP-1，MMP-2，MMP-3，MMP-7，MMP-9及MMP-14的基因表達(dá)和蛋白含量均較低，隨著胎齡的增加，表達(dá)水平逐漸升高，在妊娠晚期皮膚中這些基因的mRNA和蛋白含量都顯著增強(qiáng)(P＜0.05)。TIMP-2和TIMP-3的基因表達(dá)和蛋白含量呈相同的變化趨勢(shì)，隨著胎兒皮膚不斷發(fā)育而逐漸升高。在不同妊娠時(shí)期的胎兒皮膚中，，TIMP-1基因表達(dá)變化不顯著，但蛋白含量水平則逐漸增大，在妊娠晚期皮膚內(nèi)含量明顯高于妊娠早期(P＜0.05)。 2．在正常皮膚組織內(nèi)，MMPs和TIMPs的基因表達(dá)較弱，蛋白含量較低，蛋白陽性顆粒主要分布于表皮基底層細(xì)胞和少量的成纖維細(xì)胞中；在增殖期的瘢痕中MMPs和TIMPs基因和蛋白表達(dá)都顯著升高(P＜0.05)，在成熟期的瘢痕中，MMPs基因表達(dá)量降至正常水平，但蛋白含量明顯高于正常皮膚(P＜0.05)，蛋白
[Abstract]:Objective: to study the changes of gene expression, protein content and distribution of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinase (TIMPs) in fetal skin and hypertrophic scars at different stages of development. To explore the mechanism of scar-free healing and some mechanisms of hypertrophic scar formation and maturation. Methods: 1Fetal skin terminating pregnancy due to trauma and other unexpected factors, 18 cases of fetal back full skin, gestational age (EGA) 13 weeks. The specimens were divided into three groups according to their gestational age: early stage (13 ~ 19 weeks), middle stage (21 ~ 25 weeks) and late stage (27 ~ 33 weeks), 6 cases in each group. 2Hypertrophic scar tissue was obtained from 16 cases of hypertrophic scar from patients undergoing plastic surgery. The mean age was 26 years and the wound healing time ranged from 4 months to 11 years. Scar hyperplasia time was less than 1 year in 8 cases, 1-2 years in 4 cases, more than 2 years in 4 cases. The normal control group (n = 8) was taken from the whole skin of the donor area of the patient. After aseptic sampling, part of the samples were frozen in liquid nitrogen immediately and used for reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay to detect MMP-1,MMP-2,MMP-3,MMP-7,MMP-9,MMP-14,TIMP-1,. Changes of TIMP-2 and TIMP-3 gene expression and protein content; The other part was fixed with 10% neutral formaldehyde solution, dehydrated and embedded in paraffin. The other part was used for immunohistochemical test and observed under microscope. Results: 1. The gene expression and protein content of MMP-1,MMP-2,MMP-3,MMP-7,MMP-9 and MMP-14 in fetal skin of early pregnancy were lower, and the expression level increased gradually with the increase of fetal age. In the third trimester of pregnancy, the mRNA and protein contents of these genes were significantly increased (P < 0.05), and the gene expression and protein content of TIMP-2 and TIMP-3 showed the same change trend, and gradually increased with the development of fetal skin. The expression of TIMP-1 gene did not change significantly in fetal skin of different pregnancy stages, but the protein content increased gradually, and the content of protein in the skin of late pregnancy was significantly higher than that in the early trimester of pregnancy (P < 0.05). 2. In normal skin tissues, the gene expression of MMPs and TIMPs was weak and the protein content was low. The positive granules of protein were mainly distributed in the basal cells of epidermis and a small number of fibroblasts. The expression of MMPs and TIMPs gene and protein in proliferative scar were significantly increased (P < 0.05). In mature scar, the expression of MMPs gene decreased to normal level, but the protein content was significantly higher than that in normal skin (P < 0.05).
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R363

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