血卟啉单甲醚光动力治疗鲜红斑痣的临床和基础研究

发布时间：2018-06-10 17:25

本文选题：鲜红斑痣 + 光动力　；参考：《北京协和医学院》2011年博士论文

【摘要】：第一部分光动力、脉冲染料激光及强脉冲光治疗鲜红斑痣的临床疗效观察目的评价和比较光动力(PDT)、脉冲染料激光(PDL)和强脉冲光(IPL)治疗鲜红斑痣的临床疗效。方法PDT以血卟啉单甲醚为光敏剂,532nm连续激光为光源,PDL为595nm激光,IPL根据不同皮损选择560nm,590nm或640nm的滤光片,回顾性总结和分析不同性别、皮损类型及皮损部位与三种治疗的疗效关系。结果患者总数共130例,其中PDT患者35例,PDL患者56例,IPL患者39例。三种治疗组在性别、皮损类型及治疗部位的构成比上无统计学差异(p分别为0.904、0.929及0.987)。总体疗效PDT最高,其次为PDL,最后为IPL,三者的显效率分别为为54.3%、33.9%和20.5%；不同性别之间三种治疗的疗效无显著差异(p分别为0.225、0.821和0.145)；PDT组粉红型疗效明显优于紫红型(p=0.021)和增厚型(p=0.026),紫红型和增厚型之间疗效无差异(p=0.068)；PDL组及IPL组三种皮损类型之间疗效均无差异(PDL组p分别为0.226、0.400和0.946；IPL组p分别为0.803、0.095和0.069)；PDT和PDL组中颈部疗效优于面部(p分别为0.002和0.004),IPL组的疗效无差异(p=0.097)。结论PDT是治疗鲜红斑痣安全有效的方法,总体疗效优于PDL和IPL。第二部分血管内皮细胞和角质形成细胞对血卟啉单甲醚吸收特性的研究目的观察和比较血管内皮细胞(ECV304)和角质形成细胞(HaCaT)对光敏剂血卟啉单甲醚(HMME)的吸收特性。方法取对数生长期的ECV304和HaCaT分别与50、100、150、200、250μg/ml的HMME孵育,孵育时间为16h；将150μg/ml的HMME分别与上述两种细胞孵育,孵育时间分别为15min、30min、lh、3h、8h、12h、24h。激光扫描共聚焦显微镜检测荧光强度。结果浓度依赖性结果显示ECV304的平均荧光强度分别为74.00、125.57、135.24、141.99、132.09；HaCaT的平均荧光强度分别为93.88、102.45、112.59、108.23、10,1.70。时间依赖性结果显示ECV304的平均荧光强度分别为95.07、103.97、105.96、108.99、112.93、115.36、122.91；HaCaT的平均荧光强度分别为104.25、106.60、108.72、113.75、117.66、114.90、118.14。结论ECV304和HaCaT对HMME的吸收在一定浓度范围和时间范围内均呈孵育浓度和孵育时间依赖性。第三部分血卟啉单甲醚在血管内皮细胞和角质形成细胞中的定位及光动力治疗靶点的研究目的探讨血卟啉单甲醚(HMME)在血管内皮细胞系ECV304和角质形成细胞系HaCaT的亚细胞定位及光动力治疗的作用靶点。方法HMME与ECV304和HaCaT分别孵育1h和18h,荧光探针分别标记线粒体、核膜、细胞膜和过氧化物酶体,观察不同时间点下HMME与细胞器的结合率；HMME与ECV304和HaCaT分别孵育20h,给予532nm连续激光照射,比较照光前后上述细胞器中HMME的荧光强度变化。结果HMME与细胞器的结合率实验结果显示,ECV3041h组线粒体、核膜、细胞膜、过氧化物酶体HMME的结合率分别为1.18、0.72、0.95、0.68,18h组分别为1.35、0.83、0.73、0.91; HaCaTlh组分别为1.09、0.66、0.92、0.77,18h组分别为1.13、0.86、0.72、1.10。HMME在细胞器中的漂白率实验结果显示,ECV304分别为18.22%、10.77%、7.44%、8.56%, HaCaT分别为11.90%、5.02%、3.82%、8.90%。结论ECV304 HMME主要定位于线粒体,HaCaT定位于的线粒体和过氧化物酶体；ECV304光动力的主要作用靶点可能为线粒体,而HaCaT的主要作用靶点可能为线粒体和过氧化物酶体。
[Abstract]:The first part is photodynamic therapy, pulsed dye laser and intense pulsed light in the treatment of port wine stains.
Objective to evaluate and compare the clinical efficacy of photodynamic (PDT), pulsed dye laser (PDL) and strong pulse light (IPL) in the treatment of nevus fresh red spot. Methods PDT was used as a photosensitizer with hematoporphyrin monomethyl ether, 532nm continuous laser as light source, PDL as 595nm laser, IPL based on different skin lesions to select 560nm, 590nm or 640nm filters, and retrospective summary and analysis of different sex, The relationship between the type of skin lesion and the site of skin lesion and the curative effect of three kinds of treatment. The total number of patients was 130 cases, including 35 cases of PDT, 56 cases of PDL and 39 cases of IPL. There was no statistical difference between the three treatment groups in the sex, type of skin lesion and the position of treatment (P respectively 0.904,0.929 and 0.987). The overall effect was the highest, followed by PDL, and finally I. PL, the effective rates of the three were 54.3%, 33.9% and 20.5%, and there was no significant difference between the three treatments (P 0.225,0.821 and 0.145 respectively). The efficacy of the PDT group was obviously superior to the purple red type (p=0.021) and the thickening type (p=0.026), and there was no difference between the purple and the thickening types (p=0.068), and three types of skin lesions in PDL and IPL groups. There was no difference in the effect between the types (group PDL, P, 0.226,0.400 and 0.946, P in group IPL, 0.803,0.095 and 0.069), and in group PDT and PDL, the effect of the neck was better than that of the face (P 0.002 and 0.004 respectively), and there was no difference in the efficacy of IPL group (p=0.097). Conclusion PDT is an effective method for the treatment of nevus of fresh erythema.
The second part is about the absorption characteristics of vascular endothelial cells and keratinocytes to hematoporphyrin monomethyl ether.
Objective To observe and compare the absorption characteristics of vascular endothelial cells (ECV304) and keratinocyte (HaCaT) on the photosensitizer, hematoporphyrin monomethyl ether (HMME). Methods the logarithmic growth period ECV304 and HaCaT were incubated with HMME of 50100150200250 mu g/ml respectively, and the incubation time was 16h, and the HMME of the 150 micron /ml was incubated with the above two cells respectively. The fluorescence intensity was detected by 15min, 30min, LH, 3h, 8h, 12h, 24h. laser scanning confocal microscopy. The result of concentration dependence showed that the average fluorescence intensity of ECV304 was 74.00125.57135.24141.99132.09, and the average fluorescence intensity of HaCaT was 93.88102.45112.59108.23,10,1.70. time dependent results showing ECV304. The average fluorescence intensity is 95.07103.97105.96108.99112.93115.36122.91, and the average fluorescence intensity of HaCaT is 104.25106.60108.72113.75117.66114.90118.14. conclusion ECV304 and HaCaT HMME absorption in a certain concentration range and time range are both incubation concentration and incubation time dependence.
The third part is the localization of hematoporphyrin monomethyl ether in vascular endothelial cells and keratinocytes and the target of photodynamic therapy.
Objective to investigate the subcellular localization and photodynamic target of hematoporphyrin monomethyl ether (HMME) in vascular endothelial cell line ECV304 and keratinocyte line HaCaT. Methods HMME and ECV304 and HaCaT were incubated with 1H and 18h respectively. The fluorescent probes labeled mitochondria, nuclear membrane, cell membrane and peroxisome respectively, and observed HMME at different time points. The binding rate of organelles; HMME and ECV304 and HaCaT were incubated for 20h respectively. 532nm continuous laser irradiation was given, and the fluorescence intensity of HMME in the above-mentioned organelles before and after illumination was compared. Results the binding rate of HMME and organelles showed that the binding rate of mitochondria, nuclear membrane, cell membrane and peroxisome HMME in ECV3041h group was 1.18,0.72,0., respectively 1.18,0.72,0.. The 95,0.68,18h group was 1.35,0.83,0.73,0.91, and the results of the bleaching rate of 1.13,0.86,0.72,1.10.HMME in the group 1.09,0.66,0.92,0.77,18h group respectively showed that the ECV304 was 18.22%, 10.77%, 7.44%, 8.56%, and HaCaT were 11.90%, 5.02%, 3.82% respectively, and 8.90%. concluded that ECV304 HMME was mainly located in the mitochondria, HaCaT determination. Mitochondria and peroxisomes are located; the main target of ECV304 photodynamic may be mitochondria, and the main target of HaCaT may be mitochondria and peroxisomes.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R758.51

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