本文選題：強(qiáng)脈沖光 + 長波紫外線　； 參考：《第四軍醫(yī)大學(xué)》2011年博士論文

【摘要】：皮膚老化分為自然老化和光老化。后者主要由日光中的紫外線照射引起,特別是長波紫外線UVA的照射。對于光老化皮膚的治療經(jīng)歷了兩個(gè)階段:有創(chuàng)性治療和無創(chuàng)性治療,由于前者副作用較多,故其應(yīng)用逐漸減少。強(qiáng)脈沖光(Intense Pulsed Light IPL)技術(shù)是近年來產(chǎn)生的一種治療光老化皮膚的新的無創(chuàng)性方法,在臨床研究中己證實(shí)能有效地改善光老化皮膚的許多癥狀,所以在臨床得以普遍應(yīng)用。在強(qiáng)脈沖光照射對皮膚光老化的研究方面,雖然取得了一定的成績,但也存在分歧,并且以前的研究都是在正常組織和細(xì)胞上進(jìn)行。正常細(xì)胞與光老化細(xì)胞的情況相差甚遠(yuǎn)。在細(xì)胞生物學(xué)研究方面,目前文獻(xiàn)上還未見關(guān)于強(qiáng)脈沖光照射對細(xì)胞端粒影響的報(bào)道。 本實(shí)驗(yàn)旨在用UVA作為對照,觀察單次IPL照射對人皮膚成纖維細(xì)胞增殖活性和培養(yǎng)細(xì)胞上清液中MMP-1的表達(dá)情況。多次IPL照射,對細(xì)胞老化指標(biāo)的影響。IPL照射對PUVA誘導(dǎo)的老化成纖維細(xì)胞的影響。從而更好的理解光老化的分子機(jī)制和IPL光子嫩膚機(jī)制。 實(shí)驗(yàn)一:細(xì)胞分成7組,沒有任何照射的一組作為對照組,三組分別接受IPL5J/cm~2,15J/cm~2和20J/cm~2照射,其它三組接受UVA9J/cm~2,11J/cm~2和13J/cm~2照射。觀察不同劑量IPL和UVA照射后第二天細(xì)胞的形態(tài)和細(xì)胞的增殖活性,細(xì)胞上清液中MMP-1的表達(dá)水平。結(jié)果發(fā)現(xiàn),IPL照射后,細(xì)胞形態(tài)未見明顯改變,細(xì)胞增殖活性明顯增加,但不存在劑量依賴性,以臨床常用劑量15J/cm~2組細(xì)胞活性最高。UVA照射后,細(xì)胞變圓,皺縮,部分死亡,細(xì)胞活性下降,并且有劑量依賴性。UVA9J/cm~2照射后,細(xì)胞活性大約在80%。IPL照射后,MMP-1的分泌水平下降,以IPL15J/cm~2組下降最明顯。UVA照射后,MMP-1的分泌水平上升,但不存在劑量依賴性,以UVA9J/cm~2照射后,水平最高。我們接著把細(xì)胞分成四組:即對照組, UVA9J/cm~2照射組, IPL15J/cm~2照射組和UVA9J/cm~2+IPL15J/cm~2照射組(成纖維細(xì)胞接受9J/cm~2UVA照射后隨即給予15J/cm~2IPL照射),觀察IPL照射能否降低UVA導(dǎo)致的MMP-1的上升。結(jié)果發(fā)現(xiàn)UVA照射后再進(jìn)行IPL照射,MMP-1的水平回復(fù)到接近正常水平。 實(shí)驗(yàn)二:細(xì)胞分成三組,一組未接受照射作為對照組,其它兩組分別接受IPL15J/cm~2和UVA9J/cm~2照射,每天一次,連續(xù)照射5天。在第6天,收集細(xì)胞,分別進(jìn)行β-半乳糖苷酶染色,細(xì)胞周期,細(xì)胞內(nèi)活性氧和端粒長度的測定。結(jié)果發(fā)現(xiàn):連續(xù)5天的IPL照射后,β-半乳糖苷酶染色和端粒長度與正常對照相比,未見明顯變化(p0.05)。細(xì)胞周期中G1期所占百分比下降,細(xì)胞內(nèi)活性氧下降(p0.05);連續(xù)5次的UVA照射后,與正常對照相比,β-半乳糖苷酶染色陽性細(xì)胞的百分?jǐn)?shù)增加,細(xì)胞周期中G1期所占百分比未見明顯變化,細(xì)胞內(nèi)活性氧上升和端粒長度縮短(p0.05)。 實(shí)驗(yàn)三:細(xì)胞分成三組:一組用非毒性的8-甲氧補(bǔ)骨脂素孵育細(xì)胞16-18小時(shí),然后用UVA9J/cm~2照射(PUVA組),一組PUVA處理后,隨即進(jìn)行IPL15J/cm~2照射,以后每天進(jìn)行一次,連續(xù)5天,即PUVA+IPL組,同時(shí)設(shè)正常對照組,除每天更換培養(yǎng)液后,未給予任何處理。在細(xì)胞處理后1,3,5天后用MTT法檢測細(xì)胞活性。在處理5天后,收集細(xì)胞進(jìn)行β-半乳糖苷酶染色,細(xì)胞內(nèi)活性氧和端粒長度的測定。結(jié)果發(fā)現(xiàn):和對照組細(xì)胞相比,PUVA組和PUVA+IPL組β-半乳糖苷酶活性明顯增加。PUVA+IPL組β-半乳糖苷酶陽性成纖維細(xì)胞數(shù)明顯低于PUVA組(p0.05)。與對照組細(xì)胞活性相比,PUVA組和PUVA+IPL組的細(xì)胞活性呈時(shí)間依賴性下降,并且兩組之間未見明顯差別。與對照組細(xì)胞相比,PUVA處理組端粒長度縮短,細(xì)胞內(nèi)活性氧水平增加(p0.05),但與PUVA處理組相比,在PUVA處理后隨即給予多次的IPL照射可阻止端�？s短和降低細(xì)胞內(nèi)活性氧水平(p0.05)。 結(jié)論: (1) IPL和UVA單次照射人皮膚成纖維細(xì)胞后,細(xì)胞形態(tài),細(xì)胞活性和MMP-1的表達(dá)不同。UVA和IPL的聯(lián)合照射提示IPL可緩解UVA誘導(dǎo)的MMP-1的上調(diào),因此降低細(xì)胞外基質(zhì)蛋白的破壞作用。 (2)多次的UVA照射可誘導(dǎo)細(xì)胞出現(xiàn)老化表現(xiàn),如β-半乳糖苷酶活性增加,細(xì)胞內(nèi)活性氧增加,端粒長度縮短等。但多次IPL照射不會(huì)誘導(dǎo)細(xì)胞老化。 (3) PUVA處理可誘導(dǎo)培養(yǎng)的成纖維細(xì)胞出現(xiàn)細(xì)胞老化。但I(xiàn)PL照射通過降低活性氧和阻止端粒縮短而保護(hù)PUVA誘導(dǎo)的細(xì)胞老化。
[Abstract]:Skin aging is divided into natural aging and photoaging . The latter is mainly caused by ultraviolet irradiation in sunlight , especially the irradiation of long wave ultraviolet UVA . The treatment of photoaging skin has undergone two stages : invasive treatment and non - invasive treatment .

The purpose of this study was to observe the effect of single IPL irradiation on the proliferation activity of human skin fibroblasts and the expression of MMP - 1 in cultured cells by using UVA as control . The effects of IPL irradiation on the aging index of PUVA were observed . The molecular mechanism of photoaging and IPL photon rejuvenation mechanism were better understood .

The expression level of MMP - 1 and MMP - 1 were observed after irradiation with UVA9J / cm ~ 2 . After irradiation with UVA9J / cm ~ 2 , the activity of MMP - 1 increased , and the level of MMP - 1 was highest after irradiation with UVA9J / cm ~ 2 .

The results showed that the percentage of 尾 - galactosidase positive cells increased and the percentage of 尾 - galactosidase positive cells increased and the percentage of G1 phase in the cell cycle was not changed significantly after 5 consecutive days of UVA irradiation .

The activity of 尾 - galactosidase positive in PUVA group and PUVA + IPL group was significantly higher than that in control group ( P < 0.05 ) .

Conclusion :

( 1 ) After single exposure of IPL and UVA to human skin fibroblasts , the cell morphology , cell activity and expression of MMP - 1 were different . The combination of UVA and IPL suggested that IPL could alleviate the up - regulation of UVA - induced MMP - 1 , thus reducing the destruction of extracellular matrix protein .

( 2 ) Multiple doses of UVA irradiation could induce aging of cells , such as increased 尾 - galactosidase activity , increased intracellular reactive oxygen , shortened telomeric length , etc . However , multiple IPL irradiation did not induce cell aging .

( 3 ) PUVA treatment can induce cell aging in cultured fibroblasts , but IPL irradiation protects PUVA - induced cell aging by reducing reactive oxygen and preventing shortening of telomeric shortening .
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2011
【分類號(hào)】：R751

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相關(guān)期刊論文 前4條

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本文編號(hào)：1904525