本文選題：強(qiáng)脈沖光　切入點(diǎn)：成纖維細(xì)胞　出處：《第四軍醫(yī)大學(xué)》2011年碩士論文

【摘要】：紫外線輻射(ultraviolet radiation, UV)對(duì)正常人類皮膚的損傷包括日曬傷,光老化,免疫抑制及皮膚癌癥,其中皮膚的光損傷導(dǎo)致老化是近年來(lái)皮膚研究領(lǐng)域中的熱門課題之一。近年來(lái)新開展了光子嫩膚技術(shù),臨床療效觀察發(fā)現(xiàn),強(qiáng)脈沖光(intense pulsed light, IPL)照射能使色素斑減淡/消除、毛細(xì)血管擴(kuò)張改善/消除、細(xì)小皺紋消除,并且具有輕微的緊致皮膚作用等功效。因此本課題根據(jù)當(dāng)前熱點(diǎn)問(wèn)題,從細(xì)胞生物學(xué)層面研究強(qiáng)脈沖光對(duì)人皮膚成纖維細(xì)胞(fibroblast,FB)的生長(zhǎng)活力、細(xì)胞增殖能力以及細(xì)胞周期變化的影響并探討其可能的發(fā)生機(jī)制,為強(qiáng)脈沖光除皺嫩膚提供可能的理論依據(jù)。 我們?nèi)≌和そM織,分離培養(yǎng)原代人皮膚成纖維細(xì)胞,給以UVA I照射模擬光損傷。應(yīng)用CCK-8 (Cell Counting Kit-8)比色法、流式細(xì)胞儀(Flow Cytometry,FCM)分析技術(shù)和western blot技術(shù),觀察皮膚成纖維細(xì)胞經(jīng)強(qiáng)脈沖光照射后細(xì)胞的生長(zhǎng)活力、細(xì)胞增殖能力以及細(xì)胞周期蛋白的表達(dá)變化。結(jié)果發(fā)現(xiàn): (1)體外實(shí)驗(yàn)中,9J/cm~2劑量的UVA I照射后第48h出現(xiàn)增殖變緩,有損傷作用。以9J/cm2的劑量UVAⅠ照射后48h時(shí),即可觀察到細(xì)胞增殖活性出現(xiàn)了下降,隨后的各時(shí)間點(diǎn)內(nèi)與對(duì)照組相比,增殖活性明顯減小,P 0.05。臨床劑量IPL照射FB第72h之后,與對(duì)照組相比,細(xì)胞增殖活性明顯升高,P 0.01。FB經(jīng)UVAⅠ照射后再給IPL處理后,其增殖活性明顯優(yōu)于單獨(dú)UVAⅠ處理組, P0.05。 (2)單獨(dú)UVA I或IPL照射48h后,G1期細(xì)胞比例下降,而S期細(xì)胞比例升高。UVAⅠ照射后再給以IPL處理,其S期高于單獨(dú)UVAⅠ照射組,同時(shí)G1期細(xì)胞低于UVA I單獨(dú)照射組,p0.05。 (3)單獨(dú)UVA I照射細(xì)胞48 h后,細(xì)胞周期相關(guān)蛋白CyclinD1和CDK2的表達(dá)下降,而UVAⅠ+ IPL照射后,CyclinD1和CDK2表達(dá)雖然低于對(duì)照組,但明顯較單獨(dú)UVAⅠ照射組升高,p0.05。 本課題利用CCK-8、FCM及western blot技術(shù)檢測(cè)到了IPL可促進(jìn)成纖維細(xì)胞增殖和DNA合成,解釋了臨床上觀察到IPL的“嫩膚”現(xiàn)象,從而為強(qiáng)脈沖光面部美容除皺供理論依據(jù)。
[Abstract]:Ultraviolet radiation (ultraviolet, radiation, UV) of normal human skin damage including sunburn, photoaging, immunosuppression and skin cancer, which leads to photodamaged skin aging is one of the hot topics in the field of skin research in recent years. In recent years, launched a new IPL technology, clinical observation, intense pulsed light (intense pulsed light, IPL) irradiation can make pigmentation dodge / elimination, improve telangiectasia / eliminate fine wrinkles, eliminate, and has a slight firming skin effects. So this topic according to the hot issues, from the level of cell biology research of strong pulse light on human skin fibroblasts (fibroblast, FB) the growth activity, cell proliferation and cell cycle changes and explore the possible mechanism for the IPL rejuvenation wrinkle and provide a theoretical basis for the possible.
We take the normal foreskin tissue, primary human skin fibroblasts were isolated and cultured with UVA I irradiation simulating light damage. The application of CCK-8 (Cell Counting Kit-8) colorimetry, flow cytometry (Flow Cytometry FCM) analysis technology and Western blot technology, observation of skin fibroblasts by strong pulse cell growth vigor the light irradiation, changes of cell proliferation and cell cycle protein expression. Results:
(1) in vitro, the dose of 9J/cm~2 UVA after I irradiation 48h proliferation slow damage. In a dose of UVA 9J/cm2 1 48h after irradiation, and then observed the cell proliferation activity declined, compared with the control group at each time point in the following, the proliferation activity was significantly reduced, after P 0.05. FB 72h irradiated by IPL, compared with the control group, the cell proliferation activity was significantly increased P 0.01.FB after irradiation by UVA I give IPL treatment, the proliferation activity was superior to UVA alone treatment group 1, P0.05.
(2) after irradiation alone with UVA I or IPL, the proportion of G1 phase cells decreased, while the proportion of S phase cells increased..UVA I was treated with IPL after irradiation, and the S phase was higher than that of group I and group II, while the cells in G1 phase were lower than those in group IPL alone.
(3) after 48 h irradiation alone, the expression of cell cycle related protein CyclinD1 and CDK2 decreased, but the expression of CyclinD1 and CDK2 in UVA I + IPL group was lower than that in control group, but it was significantly higher than that in group UVA alone.
We used CCK-8, FCM and Western blot technology to detect IPL, which could promote fibroblast proliferation and DNA synthesis. It explained the phenomenon of IPL's "tender skin" in clinic, so as to provide theoretical basis for facial beauty and rhytidectomy with intense pulsed light.

【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R751

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 李曉格;李世榮;曹川;鄧向東;;強(qiáng)脈沖光對(duì)成纖維細(xì)胞增殖與周期的影響[J];中國(guó)美容醫(yī)學(xué);2007年10期

2 許陽(yáng);吳迪;駱丹;陳斌;薛筑云;;強(qiáng)脈沖光照射對(duì)皮膚成纖維細(xì)胞活性及其分泌基質(zhì)金屬蛋白酶的影響[J];中華皮膚科雜志;2007年12期

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本文編號(hào)：1691842