發(fā)布時(shí)間：2018-03-09 07:51

  本文選題：毛囊　切入點(diǎn)：真皮鞘細(xì)胞　出處：《中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院》2007年博士論文　論文類型：學(xué)位論文

【摘要】： [目的]如何促進(jìn)創(chuàng)面愈合和減少瘢痕形成是創(chuàng)傷研究領(lǐng)域的重點(diǎn)問題之一。皮膚愈合與局部毛囊數(shù)量密切相關(guān)的有趣現(xiàn)象引起了關(guān)注。現(xiàn)已證實(shí)毛囊真皮鞘細(xì)胞具有成體干細(xì)胞特性。那么，它在創(chuàng)傷愈合中扮演怎樣的角色?本課題對(duì)毛囊真皮鞘細(xì)胞在皮膚創(chuàng)傷愈合中的作用進(jìn)行了較系統(tǒng)的研究，旨在為皮膚創(chuàng)/燒傷和病理性瘢痕治療研究提供新的思路。 [方法]利用組織塊法原代培養(yǎng)大鼠觸須毛囊真皮鞘細(xì)胞，經(jīng)免疫細(xì)胞化學(xué)和免疫熒光染色進(jìn)行細(xì)胞鑒定，MTT檢測(cè)和光鏡、電鏡觀察細(xì)胞生長(zhǎng)特性和超微結(jié)構(gòu)特征；對(duì)BrdU細(xì)胞標(biāo)記方法、檢測(cè)手段和創(chuàng)面細(xì)胞移植時(shí)機(jī)進(jìn)行優(yōu)化，建立動(dòng)物實(shí)驗(yàn)?zāi)Ｐ�；通過超微結(jié)構(gòu)掃描電鏡定性觀察和瘢痕三維重建體積定量建立瘢痕評(píng)價(jià)體系；大鼠背部創(chuàng)面同種毛囊真皮鞘細(xì)胞移植后，通過病理形態(tài)觀察和圖像分析對(duì)創(chuàng)面愈合及瘢痕形成進(jìn)行動(dòng)態(tài)定性和定量評(píng)價(jià)；采用RT-PCR、免疫細(xì)胞化學(xué)、免疫熒光和免疫印跡技術(shù)分別從基因或蛋白水平檢測(cè)部分愈合相關(guān)蛋白在真皮鞘細(xì)胞中的表達(dá)。 [結(jié)果] (1)毛囊真皮鞘細(xì)胞具有的生長(zhǎng)特性和結(jié)構(gòu)特征：原代生長(zhǎng)遲緩、傳代后增殖旺盛，呈克隆樣叢狀生長(zhǎng)；細(xì)胞呈紡錘形，，胞體豐滿；核/漿比較大，核仁較多，胞膜下存在多量條帶狀致密斑。(2)建立了大鼠背部創(chuàng)面可示蹤細(xì)胞移植模型：以終濃度為10μmol/L的BrdU標(biāo)記細(xì)胞，于致傷前移植，術(shù)后14d的愈合組織中仍可見到植入細(xì)胞。(3)建立了皮膚創(chuàng)傷愈合瘢痕評(píng)價(jià)體系：以瘢痕中膠原纖維和膠原原纖維的排列作為定性指標(biāo)；瘢痕體積作為定量指標(biāo)。(4)毛囊真皮鞘細(xì)胞具有促進(jìn)皮膚創(chuàng)面愈合的作用：術(shù)后各時(shí)間點(diǎn)細(xì)胞移植組創(chuàng)面收縮率均大于對(duì)照組(P＜0.01)；術(shù)后7d移植組新生肉芽組織中見不均勻分布的標(biāo)記細(xì)胞，14d標(biāo)記細(xì)胞減少并出現(xiàn)凋亡；移植組愈合過程中肉芽組織形成、表皮再生、膠原增生、瘢痕形成和組織改建均較對(duì)照組提前，所形成的瘢痕量呈減少趨勢(shì)。(5)真皮鞘細(xì)胞的促愈機(jī)制：細(xì)胞中Ⅰ型前膠原蛋白mRNA高表達(dá)，bFGF、EGFR和TIMP1在蛋白水平較皮膚成纖維細(xì)胞高表達(dá)。 [結(jié)論]毛囊真皮鞘細(xì)胞為幼稚的間充質(zhì)細(xì)胞，具有潛在增殖能力；在皮膚創(chuàng)傷修復(fù)過程中參與肉芽組織的形成和膠原合成，具有一定的加速創(chuàng)面收縮、促進(jìn)愈合進(jìn)程和減少瘢痕形成的作用；該作用與真皮鞘細(xì)胞某些愈合相關(guān)蛋白高表達(dá)有關(guān)。揭示了毛囊真皮鞘細(xì)胞的生物學(xué)特性，為其在創(chuàng)傷治療中的應(yīng)用提供了實(shí)驗(yàn)依據(jù)。
[Abstract]:[objective] how to promote wound healing and reduce scar formation is one of the key problems in the field of trauma research. The interesting phenomenon that skin healing is closely related to the number of local hair follicles has attracted attention. It has been proved that the dermal sheath of hair follicles is fine. Cells have the properties of adult stem cells. What role does it play in wound healing? The role of dermal sheath cells of hair follicles in the healing of skin trauma was studied systematically in order to provide new ideas for the treatment of skin trauma / burn and pathological scar. [methods] the primary cultured rat hair follicle dermal sheath cells were cultured by tissue block method. The cells were identified by MTT assay and light microscopy by immunocytochemistry and immunofluorescence staining. The growth and ultrastructural characteristics of the cells were observed by electron microscope. The BrdU cell labeling method, detection method and the time of wound cell transplantation were optimized to establish animal experimental model, and the scar evaluation system was established by qualitative observation of ultrastructural scanning electron microscope and 3D reconstruction volume of scar. The wound healing and scar formation were evaluated qualitatively and quantitatively by histopathological observation and image analysis after transplantation of dermal sheath cells of hair follicle of rat back wound, using RT-PCR, immunocytochemistry, immunocytochemistry, immunocytochemistry, immunocytochemistry, immunocytochemistry, immunocytochemistry, Immunofluorescence and Western blotting were used to detect the expression of partial healing related proteins in dermal sheath cells. [results] the growth characteristics and structural characteristics of hair follicle dermis sheath cells were as follows: primary growth retardation, vigorous proliferation after passage, clone like plexiform growth, spindle-shaped cells with full cell body, large nucleus / plasma and more nucleoli. The model of traceable cells transplantation in rat back wounds was established by using the BrdU labeled cells with a final concentration of 10 渭 mol/L before injury. Implantation cells were still found in the healing tissue 14 days after operation. The evaluation system of scar healing was established. The arrangement of collagen fibers and procollagen fibers in scar was used as qualitative index. Scar volume as a quantitative index. (4) hair follicle dermal sheath cells can promote the healing of skin wounds: at all time points after operation, the wound contraction rate in the cell transplantation group was higher than that in the control group (P < 0.01), but it was not found in the granulation tissue of the transplantation group on the 7th day after operation. The uniform distribution of labeled cells decreased and apoptosis appeared on the 14th day. Granulation tissue formation, epidermal regeneration, collagen proliferation, scar formation and tissue remodeling in the transplantation group were earlier than those in the control group. The healing mechanism of dermal sheath cells was as follows: type I procollagen mRNA was highly expressed in the cells and the expression of bFGFEGFR and TIMP1 was higher than that of skin fibroblasts at the protein level. [conclusion] hair follicle dermal sheath cells are immature mesenchymal cells with potential proliferative ability, and participate in the formation of granulation tissue and collagen synthesis in the process of skin wound repair. The role of promoting healing process and reducing scar formation is related to the overexpression of some healing related proteins in dermal sheath cells. The biological characteristics of dermal sheath cells in hair follicles are revealed and the experimental basis for its application in wound therapy is provided.
【學(xué)位授予單位】：中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R363

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 楊麗;人毛囊源性間充質(zhì)樣干細(xì)胞的分離培養(yǎng)與成骨誘導(dǎo)[D];吉林大學(xué);2010年

本文編號(hào)：1587625