【摘要】：目的：建立接近人體皮膚老化過(guò)程的快速、簡(jiǎn)便、經(jīng)濟(jì)的實(shí)驗(yàn)動(dòng)物模型,結(jié)合皮膚老化的氧化應(yīng)激學(xué)說(shuō),驗(yàn)證瘦素(Leptin)對(duì)皮膚老化過(guò)程中活性氧(ROS)生成及抗氧化防御能力的影響及調(diào)控,判明瘦素在延緩皮膚衰老過(guò)程中可能發(fā)揮的作用及其機(jī)制,從而為瘦素在抗皮膚老化甚至再生受損皮膚的應(yīng)用尋找實(shí)驗(yàn)及理論依據(jù),為皮膚抗衰老再生醫(yī)學(xué)研究提供新的思路。方法：1.模擬人體皮膚衰老的內(nèi)外源誘發(fā)因素,以D-gal注射聯(lián)合UV輻照的方式,設(shè)置單一D-gal注射組、單一UV輻照組及空白對(duì)照組,從組織形態(tài)學(xué)(組織HE染色、Masson三色染色、彈力纖維染色)改變及氧化應(yīng)激分子指標(biāo)和代謝產(chǎn)物檢測(cè)等變化,分別進(jìn)行比較分析,尋找經(jīng)濟(jì)的實(shí)驗(yàn)動(dòng)物、有效的干預(yù)措施和合理的干預(yù)時(shí)間,構(gòu)建快速、經(jīng)濟(jì)、實(shí)用、周期短、更接近人體皮膚老化進(jìn)程和規(guī)律的動(dòng)物模型。2.在構(gòu)建的皮膚老化動(dòng)物模型基礎(chǔ)上,設(shè)置空白對(duì)照和試驗(yàn)對(duì)照,試驗(yàn)組分為低(5ug/g)、中(10ug/g)、高(20ug/g)三個(gè)劑量組給予瘦素腹腔內(nèi)注射,6周后取材對(duì)皮膚組織形態(tài)學(xué)和組織勻漿氧化應(yīng)激生化指標(biāo)進(jìn)行比較,研究瘦素調(diào)控氧化應(yīng)激、延緩皮膚老化的作用及可能的最適劑量。3.將瘦素溶液配制為低(5μg/ml)、中(10μg/ml)、高(20μg/ml)濃度,對(duì)培養(yǎng)的HSFs進(jìn)行預(yù)處理,之后行UV輻照,誘導(dǎo)細(xì)胞老化。檢測(cè)HSFs細(xì)胞活力、SA-β-gal活性,氧化應(yīng)激生化指標(biāo)及Western-blot測(cè)定HSFs中p67phox,PKCε和p66Shc蛋白的表達(dá)。研究瘦素延緩老化、調(diào)控氧化應(yīng)激的可能機(jī)制。結(jié)果：1.對(duì)CD-1小鼠以每日UV輻照能量611.28mJ/cm2，隔日一次,聯(lián)合D-gal劑量1mg/g/D,每日頸背部皮下注射的聯(lián)合措施,與其他干預(yù)組對(duì)比,干預(yù)后6周可見(jiàn)明顯的皮膚衰老組織形態(tài)學(xué)變化,氧化應(yīng)激水平也在6周達(dá)到相對(duì)穩(wěn)定狀態(tài),較其他干預(yù)組時(shí)間縮短、誘發(fā)老化效果更明顯。2.瘦素系統(tǒng)應(yīng)用,可明顯緩解D-gal聯(lián)合UV誘發(fā)皮膚衰老的組織學(xué)變化。瘦素在皮膚局部發(fā)揮出抗氧化作用,保護(hù)抗氧化酶、減少脂質(zhì)過(guò)氧化和膠原蛋白的降解。同時(shí)發(fā)現(xiàn),瘦素對(duì)抗皮膚老化進(jìn)程中的氧化應(yīng)激反應(yīng)有劑量選擇性,本實(shí)驗(yàn)中,中等劑量的瘦素(10ug/g)對(duì)衰老皮膚抗氧化酶的保護(hù)、拮抗脂質(zhì)過(guò)氧化和膠原蛋白降解,能發(fā)揮更好的作用(P0.05)。3.瘦素干預(yù)可以緩解UV對(duì)人成纖維細(xì)胞抗氧化酶活性的降低作用及對(duì)膜脂質(zhì)和膠原蛋白的破壞作用,可有效降低UV輻照誘導(dǎo)的成纖維細(xì)胞p67phox, PKCε和p66Shc表達(dá)增加。Leptin可以調(diào)控UV誘導(dǎo)的人成纖維細(xì)胞的氧化應(yīng)激狀態(tài),緩解甚至逆轉(zhuǎn)過(guò)氧化損傷,發(fā)揮細(xì)胞抗衰老甚至一定程度上的細(xì)胞再生作用。結(jié)論：細(xì)胞因子Leptin是一種多靶器官、多效應(yīng)的蛋白激素。本實(shí)驗(yàn)研究發(fā)現(xiàn),在動(dòng)物和細(xì)胞層面,瘦素均可以發(fā)揮出拮抗氧化的作用,延緩皮膚老化的組織學(xué)及細(xì)胞學(xué)改變。其機(jī)制可能與Leptin拮抗或清除活性氧自由基、降低氧化應(yīng)激反應(yīng)有關(guān)。
[Abstract]:Objective: to establish a rapid, simple and economical animal model of skin aging, combined with the oxidative stress theory of skin aging. To verify the effect and regulation of leptin on the production of reactive oxygen species (Ros) and anti-oxidative defense during skin aging, and to identify the role and mechanism of leptin in delaying skin aging. So it can provide experimental and theoretical basis for the application of leptin in the application of anti-aging and even regenerating damaged skin, and provide a new idea for the research of anti-aging and regenerative medicine of skin. Method 1: 1. The internal and external inducing factors of human skin senescence were simulated. With D-gal injection combined with UV irradiation, single D-gal injection group, single UV irradiation group and blank control group were divided into three groups: histomorphology (HE staining) and Masson trichrome staining. The changes of elastic fiber staining and the changes of oxidative stress molecular indexes and metabolites were compared and analyzed respectively to find economic experimental animals, effective intervention measures and reasonable intervention time to construct fast, economical, practical, and so on. Short cycle, closer to the human skin aging process and law of the animal model. 2. On the basis of the animal model of skin aging, blank control and experimental control were set up. Three dose groups of low (5ug/g), middle (10ug/g) and high (20ug/g) groups were given leptin intraperitoneal injection for 6 weeks to compare the skin histomorphology and biochemical indexes of oxidative stress of tissue homogenate, and to study the regulation of leptin on oxidative stress. Effect of delaying skin aging and possible optimal dose. 3. Leptin solution was prepared with low (5 渭 g/ml), medium (10 渭 g/ml) and high (20 渭 g/ml) concentrations. The cultured HSFs were pretreated with UV irradiation to induce cell aging. The activity of SA- 尾 -gal, the biochemical index of oxidative stress and the expression of p67phoxton-PKC 蔚 and p66Shc protein in HSFs were detected by Western-blot. To study the possible mechanism of leptin delaying aging and regulating oxidative stress. The result is 1: 1. CD-1 mice were treated with UV irradiation energy of 611.28mJ / cm ~ (-2) every other day, combined with D-gal dose of 1mg / g / D and subcutaneous injection of D-gal daily. Compared with other intervention groups, obvious changes of skin aging histomorphology were observed 6 weeks after intervention. The level of oxidative stress also reached a relatively stable state at 6 weeks, which was shorter than that of other intervention groups, and the effect of inducing aging was more obvious. 2. The application of leptin system can significantly alleviate the histological changes of skin senescence induced by D-gal combined with UV. Leptin exerts antioxidation effect, protects antioxidant enzymes, and reduces lipid peroxidation and collagen degradation. At the same time, it was found that leptin had dose-selective effects on oxidative stress during skin aging. In this experiment, moderate dose of leptin (10ug/g) protected aging skin from antioxidant enzymes and antagonized lipid peroxidation and collagen degradation. Can play a better role (P0.05). 3. Leptin intervention alleviated the decrease of antioxidant enzyme activity and the destruction of membrane lipid and collagen protein in human fibroblasts induced by UV. The expression of p67phox, PKC 蔚 and p66Shc in human fibroblasts induced by UV irradiation can be effectively reduced. Leptin can regulate the oxidative stress state of human fibroblasts induced by UV irradiation and even reverse the peroxidation injury. Give play to cell anti-aging and even to a certain extent of the role of cell regeneration. Conclusion: the cytokine leptin is a multi-target organ and multi-effect protein hormone. In this study, it was found that leptin could antagonize oxidation and delay the histological and cytological changes of aging skin at both animal and cell levels. The mechanism may be related to leptin antagonizing or scavenging reactive oxygen free radicals and reducing oxidative stress.
【學(xué)位授予單位】：中國(guó)人民解放軍醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R339.38

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 林筱;;導(dǎo)致皮膚老化基因及8大主因[J];科學(xué)大觀園;2009年19期

2 蘆源,張鷗;皮膚老化及其防治研究進(jìn)展[J];中國(guó)中醫(yī)基礎(chǔ)醫(yī)學(xué)雜志;2001年08期

3 ;德國(guó)研究發(fā)現(xiàn)女性皮膚老化比男性快[J];中國(guó)西部科技;2006年24期

4 ;你知道嗎?[J];百科知識(shí);2013年18期

5 朱焱;宋智琦;趙春暉;鄒偉;;雌激素受體在抗皮膚老化過(guò)程中的多效性[J];生物技術(shù)通訊;2012年04期

6 卜平元;彭程;尹朝奇;羅成群;賀全勇;毛平;;強(qiáng)脈沖光聯(lián)合MEBT/MEBO治療皮膚老化的臨床觀察[J];激光生物學(xué)報(bào);2011年05期

7 王春波,賀孟泉,秦守哲,張良,王麗君,仲偉珍,王玉貞,張建;海洋肽抗皮膚老化作用的實(shí)驗(yàn)研究[J];中華醫(yī)學(xué)美容雜志;1998年04期

8 程誠(chéng);;夏日的一抹清涼[J];青年科學(xué);2007年07期

9 ;植物提取物具有防皺功效[J];大眾科技;2008年02期

10 孫明霞 ,湯浩;具有美容功效的食物[J];科技信息;1997年05期

相關(guān)會(huì)議論文 前10條

1 劉仲榮;羅曉艷;楊慧蘭;;皮膚老化的評(píng)價(jià)及分子標(biāo)記[A];2006中國(guó)中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會(huì)議論文匯編[C];2006年

2 甄雅賢;;非創(chuàng)傷性技術(shù)在研究皮膚老化方面的應(yīng)用[A];中華醫(yī)學(xué)會(huì)第14次全國(guó)皮膚性病學(xué)術(shù)年會(huì)論文匯編[C];2008年

3 劉仲榮;楊慧蘭;;皮膚老化與線粒體DNA突變[A];2009全國(guó)中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會(huì)議論文匯編[C];2009年

4 李文海;陳周;張建中;;皮膚老化的新形式—熱老化[A];2010全國(guó)中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會(huì)議論文匯編[C];2010年

5 張敬東;;皮膚老化及防治對(duì)策[A];2002中國(guó)中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會(huì)議論文匯編[C];2002年

6 Jean-Francois Nicolay;Christophe Paillet;;假二肽的目標(biāo):抗環(huán)境(污染)刺激,抗皮膚老化,增強(qiáng)皮膚的自然保護(hù)潛能[A];2002年中國(guó)化妝品學(xué)術(shù)研討會(huì)論文集[C];2002年

7 劉仲榮;楊慧蘭;李雪梅;樊建勇;;皮膚老化的防治[A];2006中國(guó)中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會(huì)議論文匯編[C];2006年

8 龔麗萍;;皮膚老化的預(yù)防與治療新動(dòng)態(tài)[A];江西省第五次中西醫(yī)結(jié)合皮膚科學(xué)術(shù)交流會(huì)論文集[C];2006年

9 S．Nouveau-Richard;Z．Yang;S．Mac-Mary;L．Li;P.Bastien;I．Tardy;C．Bouillon;P.Humbert;O．de Lacharrie're;;皮膚老化:中國(guó)人和歐洲人的比較一項(xiàng)探索性研究[A];2006年中國(guó)化妝品學(xué)術(shù)研討會(huì)論文集[C];2006年

10 楊慧蘭;劉仲榮;;抗皮膚老化化妝品活性成分研究進(jìn)展[A];2009全國(guó)中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會(huì)議論文匯編[C];2009年

相關(guān)重要報(bào)紙文章 前10條

1 晴天;追根究底 阻擊皮膚老化[N];保健時(shí)報(bào);2004年

2 南昌大學(xué)第二附屬醫(yī)院皮膚科 教授 劉志剛;延緩皮膚老化 減少歲月留痕[N];家庭醫(yī)生報(bào);2009年

3 ;新技術(shù)延緩皮膚老化[N];中華工商時(shí)報(bào);2003年

4 宇帆;辣椒減肥是非說(shuō)[N];醫(yī)藥經(jīng)濟(jì)報(bào);2003年

5 首都醫(yī)科大學(xué)宣武醫(yī)院皮膚性病科主任 連石;美容效果也能檢測(cè)啦[N];健康報(bào);2008年

6 劉霞　何屹;你的皺紋 基因的作品[N];科技日?qǐng)?bào);2009年

7 曉麗;科技美容：麗人新法寶[N];四川日?qǐng)?bào);2001年

8 洪秀;護(hù)手，專家為你支招[N];大眾衛(wèi)生報(bào);2002年

9 白云水;老人長(zhǎng)色斑是病嗎[N];健康報(bào);2004年

10 玉仁;老年斑會(huì)癌變嗎？[N];大眾衛(wèi)生報(bào);2004年

相關(guān)博士學(xué)位論文 前3條

1 陳愛(ài)軍;皮膚老化過(guò)程中角質(zhì)形成細(xì)胞比較蛋白質(zhì)組分析與鑒定[D];重慶醫(yī)科大學(xué);2009年

2 周志強(qiáng);瘦素對(duì)皮膚哀老氧化應(yīng)激水平影響的實(shí)驗(yàn)研究[D];中國(guó)人民解放軍醫(yī)學(xué)院;2016年

3 錢(qián)革;我國(guó)部分地區(qū)成人面部皮膚老化相關(guān)因素及香煙煙霧提取物與尼古丁致皮膚老化作用和機(jī)制的研究[D];中國(guó)協(xié)和醫(yī)科大學(xué);2007年

相關(guān)碩士學(xué)位論文 前4條

1 白云;鐵蓄積對(duì)去勢(shì)大鼠內(nèi)源性皮膚老化影響的實(shí)驗(yàn)研究[D];蘇州大學(xué);2015年

2 王斌;IPL治療皮膚老化相關(guān)作用機(jī)制的研究[D];中南大學(xué);2008年

3 劉娜;日光暴露對(duì)皺紋特征的影響[D];同濟(jì)大學(xué);2005年

4 高倩;手背部皮膚紋理老化定量評(píng)價(jià)方法研究[D];中國(guó)醫(yī)科大學(xué);2006年

，

本文編號(hào)：2129362