卡泊三醇软膏预处理后局部光动力疗法对裸鼠正常皮肤损伤效应研究

发布时间：2018-04-05 12:00

本文选题：5-氨基酮戊酸　切入点：原卟啉IX　出处：《桂林医学院》2017年硕士论文

【摘要】：第一部分卡泊三醇预处理对5-氨基酮戊酸局部外用裸鼠正常皮肤后PpIX含量影响的研究目的:探讨卡泊三醇预处理裸鼠正常皮肤后,对外用5-氨基酮戊酸(5-ALA)产生原卟啉IX(PpIX)的影响。方法:以裸鼠为模型,分为ALA组、卡泊三醇(calcipotriol,CAL)联合ALA组、CAL组及空白对照组。CAL+ALA组卡泊三醇软膏预先外用3天,3天后前两组均外敷ALA,之后在不同时间点使用倒置荧光相差显微镜观察皮肤组织中PpIX的荧光强度及分布,并使用荧光分光光度计定量测定裸鼠正常皮肤组织中合成的PpIX。结果:在裸鼠组织内,ALA吸收后产生的PpIX浓度随时间逐渐增加,ALA组在5h达到高峰,CAL+ALA组在6h达高峰,在5、6、7小时CAL+ALA组PpIX含量明显高于单纯ALA组,差异有统计学意义(均p0.05)。冰冻切片显示PpIX荧光弥漫性分布于表皮层和真皮层,CAL+ALA组荧光强度在4-7小时高于ALA组。结论:CAL预处理可以提高ALA作用裸鼠正常皮肤后表皮及真皮层内PpIX含量,为临床上CAL作为局部ALA-PDT的增效剂提供了理论依据。第二部分卡泊三醇软膏预处理后局部ALA-PDT对裸鼠正常皮肤的损伤效应研究目的:本实验在上述实验基础上,进一步研究卡泊三醇软膏联合局部ALA-PDT对裸鼠正常皮肤组织的损伤效应,以明确卡泊三醇软膏预处理是否可增强PDT的作用效果,为临床治疗提供理论依据。方法:以裸鼠作为实验对象,选取42只裸鼠,随机分为3组。分别为A组:ALA-PDT组,18只;B组:CAL联合ALA-PDT组,18只;C组:空白对照组,6只。分别在实验后1-7d动态观察裸鼠上皮损伤后形态变化和24h、7d、14d组织病理变化。结果:A组裸鼠皮肤实验区域出现不同程度红斑、未见痂皮,B组裸鼠皮肤在红斑基础上出现痂皮甚至厚出血痂,C组未见异常;两组镜下表皮、真皮层均可见核损伤、坏死,胶原纤维增多,毛细血管减少等,而B组程度均大于A组,而且B组真皮层厚度大于A组。结论:卡泊三醇软膏联合ALA-PDT组较单纯应用ALA-PDT组对裸鼠正常皮肤组织有更明显的损伤效应。
[Abstract]:Part I effects of carpotriol pretreatment on PpIX content in normal skin of nude mice treated with 5-aminolevulinic acid objective: to investigate the effect of carpotriol preconditioning on the production of protoporphyrin IX-PpIX (5-ALA-5-ALA-5-aminoketovalerate) in normal skin of nude mice.Methods: nude mice were divided into ALA group.Carpotriol calcium triolol cale (CAL) combined with ALA group and control group. CAL ALA group: carpotriol ointment was pretreated for 3 days and 3 days later, both groups were treated with ALAs, and then the PpIX in skin tissue was observed by inverted fluorescence phase contrast microscope at different time points.Fluorescence intensity and distribution,The synthesis of PpIX in normal skin tissue of nude mice was determined by fluorescence spectrophotometer.Results: the concentration of PpIX produced by Ala absorption in nude mice tissues increased gradually with time. The concentration of PpIX in CAL ALA group was significantly higher than that in ALA group at 6 h after 5 hours and 7 hours in CAL ALA group (all p0.05). The results showed that the concentration of PpIX in ALA group was significantly higher than that in control group (P < 0.05). The concentration of PpIX in CAL ALA group was significantly higher than that in ALA group (P < 0.05).Frozen sections showed that the fluorescence intensity of PpIX in ALA group was higher than that in ALA group in 4-7 hours.Conclusion ALA pretreatment can increase the content of PpIX in the epidermis and dermis of normal skin of nude mice, which provides a theoretical basis for CAL as a synergist of local ALA-PDT in clinic.The second part of the study on the damage effect of local ALA-PDT on normal skin of nude mice after pretreatment with carpotriol ointment objective: to further study the damage effect of carpotriol ointment combined with local ALA-PDT on normal skin tissue of nude mice on the basis of the above experiments.To determine whether the pretreatment of carpotriol ointment can enhance the effect of PDT and provide theoretical basis for clinical treatment.Methods: 42 nude mice were randomly divided into 3 groups.Group A: ALA-PDT group (n = 18), group B (n = 18) combined with ALA-PDT group (n = 18), group C (n = 18): control group (n = 6).The morphologic changes and histopathological changes of nude mice were observed at 1-7 days after the experiment and 14 days after 24 hours of injury.Results different degrees of erythema appeared in the experimental area of the skin of the nude mice in group A, and the skin of group B was not abnormal on the basis of erythema, the epidermis and the dermis of both groups were damaged and necrosed under microscope, and the skin of group B was not abnormal on the basis of erythema.Collagen fibers increased and capillaries decreased, while the degree of group B was greater than that of group A, and the thickness of dermis in group B was greater than that in group A.Conclusion: carpotriol ointment combined with ALA-PDT has more obvious damage effect on normal skin tissue of nude mice than that of ALA-PDT alone.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R751.05

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