【摘要】： 毛囊是皮膚重要的附屬器官,具有周期性生長(zhǎng)的特點(diǎn)。目前已知毛囊干細(xì)胞位于外根鞘的隆突(bulge)區(qū),研究證明毛囊bulge細(xì)胞具有多向分化的能力,不僅能分化形成毛囊,還能夠形成皮脂腺和表皮角朊細(xì)胞。由于表皮、毛囊和皮脂腺關(guān)系密切,一般將三者統(tǒng)歸為表皮-毛囊-皮脂腺單位(epidermo- piosebaceous unit)或表皮和毛囊皮脂腺單位(piosebaceous unit)。 皮脂腺(sebaceous gland)是皮膚重要的附屬結(jié)構(gòu),具有抗炎、抵抗外來微生物的作用;它能夠抗氧化,特別是抗紫外線的侵襲,對(duì)保持皮膚的完整和功能正常有著重要的作用。皮脂腺細(xì)胞的發(fā)育分化過程十分復(fù)雜,包括細(xì)胞形態(tài)的改變和基因表達(dá)的變化。過氧化物酶體增殖物激活受體γ(peroxisome proliferator-activated receptor, PPARγ)在成脂細(xì)胞分化過程中起著關(guān)鍵的作用,它能有效促進(jìn)脂質(zhì)合成。在PPARγ的三種亞型中,PPARγ2與脂肪和皮脂腺細(xì)胞的分化尤其相關(guān)。PPARγ2是脂肪細(xì)胞分化過程中重要的調(diào)節(jié)因子,它可促使成纖維細(xì)胞或骨髓間充質(zhì)干細(xì)胞向脂肪細(xì)胞分化。盡管多項(xiàng)證據(jù)表明皮脂腺的更新和維持來源于毛囊的Bulge細(xì)胞,但是,至今仍未在體外實(shí)驗(yàn)中觀察到bulge細(xì)胞分化為皮脂腺細(xì)胞的直接證據(jù)。毛囊干細(xì)胞體外分離培養(yǎng)技術(shù)的相對(duì)滯后,影響了研究的進(jìn)一步深入。我們?cè)隗w外分離培養(yǎng)了毛囊bulge細(xì)胞,并在一定的條件下誘導(dǎo)bulge細(xì)胞向皮脂腺細(xì)胞定向分化,同時(shí)構(gòu)建PPARγ2質(zhì)粒,通過脂質(zhì)體將其轉(zhuǎn)染到bulge細(xì)胞中,觀察它對(duì)bulge細(xì)胞定向分化為皮脂腺過程中的作用。主要結(jié)果如下: 1、毛囊bulge細(xì)胞的分離培養(yǎng)及生物學(xué)特性的研究 采用免疫組化及原位雜交技術(shù),檢測(cè)大鼠毛囊Bulge區(qū)細(xì)胞干細(xì)胞表面標(biāo)志物和分化標(biāo)志物的表達(dá)情況,運(yùn)用顯微分離及酶消化法分離并培養(yǎng)大鼠毛囊Bulge細(xì)胞,觀察培養(yǎng)的毛囊bulge細(xì)胞體外生長(zhǎng)特性,同時(shí)比較了不同培養(yǎng)體系對(duì)bulge細(xì)胞生物學(xué)特性的影響。結(jié)果發(fā)現(xiàn):(1)干細(xì)胞表面標(biāo)志物K19的表達(dá)主要位于毛囊的Bulge區(qū),而該區(qū)不表達(dá)毛囊上皮的細(xì)胞分化標(biāo)志K10。(2)經(jīng)酶消化后的毛囊Bulge區(qū)組織易貼壁,24h后即有細(xì)胞爬出,沿Bulge區(qū)向外擴(kuò)展生長(zhǎng),3-4天后細(xì)胞出現(xiàn)融合,
[Abstract]:Hair follicle is an important subsidiary organ of the skin, which has the characteristic of periodic growth. Hair follicle stem cells are known to be located in the protuberant (bulge) region of the outer root sheath. It has been demonstrated that hair follicle bulge cells have the ability to differentiate into hair follicles and sebaceous glands and epidermal keratinocytes. Because the epidermis, hair follicle and sebaceous gland are closely related, they are generally classified as epidermal hair follicle-sebaceous unit (epidermo- piosebaceous unit) or epidermis and hair follicle sebaceous gland unit (piosebaceous unit). Sebaceous gland (sebaceous gland) is an important subsidiary structure of skin, which has the function of anti-inflammation and resisting foreign microorganism, and it can resist the invasion of ultraviolet rays, and it has important function to maintain the integrity and function of skin. The process of development and differentiation of sebaceous gland cells is very complicated, including the changes of cell morphology and gene expression. Peroxisome proliferator activated receptor 緯 (peroxisome proliferator-activated receptor, PPAR 緯) plays a key role in the process of adipoblast differentiation, which can effectively promote lipid synthesis. Among the three subtypes of PPAR 緯, PPAR緯 2 is especially related to the differentiation of adipose and sebaceous gland cells. PPAR- 緯 2 is an important regulatory factor in adipocyte differentiation, which can induce fibroblasts or bone marrow mesenchymal stem cells to differentiate into adipocytes. Although many evidences suggest that the regeneration and maintenance of sebaceous glands originate from Bulge cells in hair follicles, there is no direct evidence that bulge cells differentiate into sebaceous gland cells in vitro. The technology of hair follicle stem cell isolation and culture in vitro is lagging behind, which affects the further research. Hair follicle bulge cells were isolated and cultured in vitro, and bulge cells were induced to differentiate into sebaceous gland cells under certain conditions. At the same time, PPAR 緯 2 plasmid was constructed and transfected into bulge cells by liposome. To observe its effect on the differentiation of bulge cells into sebaceous glands. The main results are as follows: 1. Isolation, culture and biological characteristics of hair follicle bulge cells Immunohistochemistry and in situ hybridization, The expression of stem cell surface markers and differentiation markers in Bulge region of rat hair follicle was detected. The Bulge cells of hair follicle were isolated and cultured by microdissection and enzyme digestion. The growth characteristics of cultured bulge cells were observed in vitro. The effects of different culture systems on the biological characteristics of bulge cells were also compared. The results showed that: (1) the expression of stem cell surface marker K19 was mainly located in the Bulge region of the hair follicle, but the cell differentiation marker of the hair follicle epithelium was not expressed in this area. (2) the tissue of the Bulge region of the hair follicle easily adhered to the wall 24 hours after the enzyme digestion. After 3 to 4 days of growth along the Bulge region, the cells were fused.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R329

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