优化ALA-PDT的封包时间对豚鼠银屑病模型生物效应的影响
发布时间：2018-09-12 06:25
【摘要】：目的：1.通过采用局部不同封包时间的ALA-PDT治疗豚鼠耳背部寻常型银屑病模型,观察并研究其对局部皮损中PpⅨ含量、表皮细胞的增殖、CD4+和CD8+T细胞的表达与生物效应的影响。2.选择最佳ALA-PDT治疗参数,优化其治疗方案,为临床应用提供理论基础和指导性意见。 方法：采用局部涂抹心得安的方法建立豚鼠耳寻常型银屑病模型；应用荧光分光光度计分别检测ALA经不同的封包时间(1h、3h、15h)后皮损组织中PpⅨ的含量；采用波长为635nm半导体激光(输出功率30mW,垂直照射5 min,能量密度为12J/cm2)对各封包组进行PDT治疗；各组分别在实验前及实验后第7d、14d、21d、28d进行肉眼观察皮损的愈合情况,通过HE染色观察组织病理学改变,通过免疫组化法染色观察表皮细胞增殖(PCNA)及CD4+和CD8+T细胞浸润。统计方法采用T检验及析因设计的方差分析,结果采用SPSS18.0进行统计学处理。 结果：1.荧光分光光度计检测结果显示：在15h内,局部皮损中PpⅨ含量随着封包时间的延长而升高,且在封包后1h-3h产生的速率最快,封包3h时可达到15h的63%；而3h-15h虽然PpⅨ含量逐渐升高,但速率最慢。2.肉眼及组织病理学观察结果显示：各ALA封包组经PDT治疗后与模型对照组比较均有显著疗效,其组织病理学评分统计学显示有显著差异。其中,ALA封包15h组皮损的组织病理学改善最快,痊愈所需时间最短,其次是ALA封包3h组,再次是ALA封包1h组。3.免疫组化结果显示：各ALA封包组表皮中的PCNA、CD4+及CD8+表达率在治疗前后统计学均有显著差异。其中,ALA封包15h组PCNA、CD4+及CD8+表达率降低最快,其次是ALA封包3h组,再次是ALA封包1h组。4.ALA封包15h组痊愈后有30%的动物局部皮损处出现色素沉着,而其他组动物则无上述表现。 结论：1.局部ALA-PDT治疗豚鼠耳背部寻常型银屑病模型有显著的疗效,且局部皮损的所需的痊愈时间与ALA封包时间的长短成负相关。2.ALA封包时间的长短与局部皮损中PpⅨ含量成正相关,且ALA封包后1h-3h生成PpⅨ的速率最快。3.局部ALA-PDT可抑制表皮细胞的过度增殖,减少CD4+、CD8+T细胞的浸润,其作用程度与局部PpIX的含量成正相关。
[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.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R-332;R758.63
本文编号：2238172
[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.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R-332;R758.63
【参考文献】
相关期刊论文 前2条
1 杨洪浦;银屑病的中医治法概述[J];现代中西医结合杂志;2004年05期
2 李康英;段世军;刘宏伟;谢光辉;;5-氨基酮戊酸-光动力疗法治疗豚鼠银屑病模型的实验研究[J];中国激光医学杂志;2010年06期
，本文编号：2238172
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