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優(yōu)化ALA-PDT的封包時(shí)間對(duì)豚鼠銀屑病模型生物效應(yīng)的影響

發(fā)布時(shí)間:2018-09-12 06:25
【摘要】:目的:1.通過(guò)采用局部不同封包時(shí)間的ALA-PDT治療豚鼠耳背部尋常型銀屑病模型,觀察并研究其對(duì)局部皮損中PpⅨ含量、表皮細(xì)胞的增殖、CD4+和CD8+T細(xì)胞的表達(dá)與生物效應(yīng)的影響。2.選擇最佳ALA-PDT治療參數(shù),優(yōu)化其治療方案,為臨床應(yīng)用提供理論基礎(chǔ)和指導(dǎo)性意見(jiàn)。 方法:采用局部涂抹心得安的方法建立豚鼠耳尋常型銀屑病模型;應(yīng)用熒光分光光度計(jì)分別檢測(cè)ALA經(jīng)不同的封包時(shí)間(1h、3h、15h)后皮損組織中PpⅨ的含量;采用波長(zhǎng)為635nm半導(dǎo)體激光(輸出功率30mW,垂直照射5 min,能量密度為12J/cm2)對(duì)各封包組進(jìn)行PDT治療;各組分別在實(shí)驗(yàn)前及實(shí)驗(yàn)后第7d、14d、21d、28d進(jìn)行肉眼觀察皮損的愈合情況,通過(guò)HE染色觀察組織病理學(xué)改變,通過(guò)免疫組化法染色觀察表皮細(xì)胞增殖(PCNA)及CD4+和CD8+T細(xì)胞浸潤(rùn)。統(tǒng)計(jì)方法采用T檢驗(yàn)及析因設(shè)計(jì)的方差分析,結(jié)果采用SPSS18.0進(jìn)行統(tǒng)計(jì)學(xué)處理。 結(jié)果:1.熒光分光光度計(jì)檢測(cè)結(jié)果顯示:在15h內(nèi),局部皮損中PpⅨ含量隨著封包時(shí)間的延長(zhǎng)而升高,且在封包后1h-3h產(chǎn)生的速率最快,封包3h時(shí)可達(dá)到15h的63%;而3h-15h雖然PpⅨ含量逐漸升高,但速率最慢。2.肉眼及組織病理學(xué)觀察結(jié)果顯示:各ALA封包組經(jīng)PDT治療后與模型對(duì)照組比較均有顯著療效,其組織病理學(xué)評(píng)分統(tǒng)計(jì)學(xué)顯示有顯著差異。其中,ALA封包15h組皮損的組織病理學(xué)改善最快,痊愈所需時(shí)間最短,其次是ALA封包3h組,再次是ALA封包1h組。3.免疫組化結(jié)果顯示:各ALA封包組表皮中的PCNA、CD4+及CD8+表達(dá)率在治療前后統(tǒng)計(jì)學(xué)均有顯著差異。其中,ALA封包15h組PCNA、CD4+及CD8+表達(dá)率降低最快,其次是ALA封包3h組,再次是ALA封包1h組。4.ALA封包15h組痊愈后有30%的動(dòng)物局部皮損處出現(xiàn)色素沉著,而其他組動(dòng)物則無(wú)上述表現(xiàn)。 結(jié)論:1.局部ALA-PDT治療豚鼠耳背部尋常型銀屑病模型有顯著的療效,且局部皮損的所需的痊愈時(shí)間與ALA封包時(shí)間的長(zhǎng)短成負(fù)相關(guān)。2.ALA封包時(shí)間的長(zhǎng)短與局部皮損中PpⅨ含量成正相關(guān),且ALA封包后1h-3h生成PpⅨ的速率最快。3.局部ALA-PDT可抑制表皮細(xì)胞的過(guò)度增殖,減少CD4+、CD8+T細(xì)胞的浸潤(rùn),其作用程度與局部PpIX的含量成正相關(guān)。
[Abstract]:AIM: 1. To observe and study the effects of ALA-PDT on the contents of Pp_, the proliferation of epidermal cells, the expression of CD4 + and CD8 + T cells and the biological effects in the local lesions of guinea pigs with auricular and dorsal psoriasis vulgaris. 2. To optimize the therapeutic regimen of ALA-PDT by choosing the best therapeutic parameters. For theoretical basis and guiding opinions.
METHODS: The model of psoriasis vulgaris in guinea pig ears was established by topical application of propranolol, and the content of Pp_in the lesions of ALA after different encapsulation time (1h, 3h, 15h) was detected by fluorescence spectrophotometer, respectively. PDT was performed in the encapsulation group, and the healing of the lesions was observed with naked eyes before and 7, 14, 21 and 28 days after the experiment. The histopathological changes were observed by HE staining, and the proliferation of epidermal cells (PCNA) and the infiltration of CD4 + and CD8 + T cells were observed by immunohistochemical staining. The results were statistically analyzed by SPSS18.0.
Results: 1. The results of fluorescence spectrophotometer showed that the content of Pp_in local skin lesions increased with the encapsulation time within 15 hours, and the fastest rate was found at 1-3 hours after encapsulation, and reached 63% at 3 hours after encapsulation, while the content of Pp_gradually increased at 3-15 hours, but the slowest rate was found at 3-15 hours after encapsulation. After PDT treatment, the histopathological score of ALA group was significantly different from that of the model control group. Among them, ALA group had the fastest histopathological improvement and the shortest healing time, followed by ALA group for 3 hours and ALA group for 1 hour. The expression rates of PCNA, CD4 + and CD8 + in the epidermis of ALA group were significantly different before and after treatment. The expression rates of PCNA, CD4 + and CD8 + decreased fastest in ALA group at 15h, followed by ALA group at 3H and ALA group at 1h. Describe the performance.
CONCLUSIONS: 1. Local ALA-PDT is effective in treating guinea pig ear and back psoriasis vulgaris model, and the healing time of local lesions is negatively correlated with the time of ALA encapsulation. 2. The time of ALA encapsulation is positively correlated with the content of Pp_in local lesions, and the rate of Pp_formation is the fastest 1-3 hours after ALA encapsulation. It can inhibit the excessive proliferation of epidermal cells and reduce the infiltration of CD4 +, CD8 + T cells. The degree of its effect is positively correlated with the local PpIX content.
【學(xué)位授予單位】:暨南大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2011
【分類號(hào)】:R-332;R758.63

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 楊洪浦;銀屑病的中醫(yī)治法概述[J];現(xiàn)代中西醫(yī)結(jié)合雜志;2004年05期

2 李康英;段世軍;劉宏偉;謝光輝;;5-氨基酮戊酸-光動(dòng)力療法治療豚鼠銀屑病模型的實(shí)驗(yàn)研究[J];中國(guó)激光醫(yī)學(xué)雜志;2010年06期



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