强脉冲光照射对UVAⅠ诱导成纤维细胞光损伤的保护作用研究

发布时间：2018-03-31 18:06

本文选题：强脉冲光　切入点：成纤维细胞　出处：《第四军医大学》2011年硕士论文

【摘要】：紫外线辐射(ultraviolet radiation, UV)对正常人类皮肤的损伤包括日晒伤,光老化,免疫抑制及皮肤癌症,其中皮肤的光损伤导致老化是近年来皮肤研究领域中的热门课题之一。近年来新开展了光子嫩肤技术,临床疗效观察发现,强脉冲光(intense pulsed light, IPL)照射能使色素斑减淡/消除、毛细血管扩张改善/消除、细小皱纹消除,并且具有轻微的紧致皮肤作用等功效。因此本课题根据当前热点问题,从细胞生物学层面研究强脉冲光对人皮肤成纤维细胞(fibroblast,FB)的生长活力、细胞增殖能力以及细胞周期变化的影响并探讨其可能的发生机制,为强脉冲光除皱嫩肤提供可能的理论依据。我们取正常儿童包皮组织,分离培养原代人皮肤成纤维细胞,给以UVA I照射模拟光损伤。应用CCK-8 (Cell Counting Kit-8)比色法、流式细胞仪(Flow Cytometry,FCM)分析技术和western blot技术,观察皮肤成纤维细胞经强脉冲光照射后细胞的生长活力、细胞增殖能力以及细胞周期蛋白的表达变化。结果发现: (1)体外实验中,9J/cm~2剂量的UVA I照射后第48h出现增殖变缓,有损伤作用。以9J/cm2的剂量UVAⅠ照射后48h时,即可观察到细胞增殖活性出现了下降,随后的各时间点内与对照组相比,增殖活性明显减小,P 0.05。临床剂量IPL照射FB第72h之后,与对照组相比,细胞增殖活性明显升高,P 0.01。FB经UVAⅠ照射后再给IPL处理后,其增殖活性明显优于单独UVAⅠ处理组, P0.05。 (2)单独UVA I或IPL照射48h后,G1期细胞比例下降,而S期细胞比例升高。UVAⅠ照射后再给以IPL处理,其S期高于单独UVAⅠ照射组,同时G1期细胞低于UVA I单独照射组,p0.05。 (3)单独UVA I照射细胞48 h后,细胞周期相关蛋白CyclinD1和CDK2的表达下降,而UVAⅠ+ IPL照射后,CyclinD1和CDK2表达虽然低于对照组,但明显较单独UVAⅠ照射组升高,p0.05。本课题利用CCK-8、FCM及western blot技术检测到了IPL可促进成纤维细胞增殖和DNA合成,解释了临床上观察到IPL的“嫩肤”现象,从而为强脉冲光面部美容除皱供理论依据。
[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.

【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R751

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