本文選題：5-氨基酮戊酸　切入點：原卟啉IX　出處：《桂林醫(yī)學院》2017年碩士論文

【摘要】：第一部分卡泊三醇預處理對5-氨基酮戊酸局部外用裸鼠正常皮膚后PpIX含量影響的研究目的:探討卡泊三醇預處理裸鼠正常皮膚后,對外用5-氨基酮戊酸(5-ALA)產(chǎn)生原卟啉IX(PpIX)的影響。方法:以裸鼠為模型,分為ALA組、卡泊三醇(calcipotriol,CAL)聯(lián)合ALA組、CAL組及空白對照組。CAL+ALA組卡泊三醇軟膏預先外用3天,3天后前兩組均外敷ALA,之后在不同時間點使用倒置熒光相差顯微鏡觀察皮膚組織中PpIX的熒光強度及分布,并使用熒光分光光度計定量測定裸鼠正常皮膚組織中合成的PpIX。結(jié)果:在裸鼠組織內(nèi),ALA吸收后產(chǎn)生的PpIX濃度隨時間逐漸增加,ALA組在5h達到高峰,CAL+ALA組在6h達高峰,在5、6、7小時CAL+ALA組PpIX含量明顯高于單純ALA組,差異有統(tǒng)計學意義(均p0.05)。冰凍切片顯示PpIX熒光彌漫性分布于表皮層和真皮層,CAL+ALA組熒光強度在4-7小時高于ALA組。結(jié)論:CAL預處理可以提高ALA作用裸鼠正常皮膚后表皮及真皮層內(nèi)PpIX含量,為臨床上CAL作為局部ALA-PDT的增效劑提供了理論依據(jù)。第二部分卡泊三醇軟膏預處理后局部ALA-PDT對裸鼠正常皮膚的損傷效應研究目的:本實驗在上述實驗基礎上,進一步研究卡泊三醇軟膏聯(lián)合局部ALA-PDT對裸鼠正常皮膚組織的損傷效應,以明確卡泊三醇軟膏預處理是否可增強PDT的作用效果,為臨床治療提供理論依據(jù)。方法:以裸鼠作為實驗對象,選取42只裸鼠,隨機分為3組。分別為A組:ALA-PDT組,18只;B組:CAL聯(lián)合ALA-PDT組,18只;C組:空白對照組,6只。分別在實驗后1-7d動態(tài)觀察裸鼠上皮損傷后形態(tài)變化和24h、7d、14d組織病理變化。結(jié)果:A組裸鼠皮膚實驗區(qū)域出現(xiàn)不同程度紅斑、未見痂皮,B組裸鼠皮膚在紅斑基礎上出現(xiàn)痂皮甚至厚出血痂,C組未見異常;兩組鏡下表皮、真皮層均可見核損傷、壞死,膠原纖維增多,毛細血管減少等,而B組程度均大于A組,而且B組真皮層厚度大于A組。結(jié)論:卡泊三醇軟膏聯(lián)合ALA-PDT組較單純應用ALA-PDT組對裸鼠正常皮膚組織有更明顯的損傷效應。
[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.
【學位授予單位】：桂林醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R751.05

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