中波高能紫外光誘導(dǎo)豚鼠皮膚色素沉著改變的實(shí)驗(yàn)研究
發(fā)布時(shí)間：2018-12-30 21:19
【摘要】：目的通過(guò)研究中波高能紫外光對(duì)豚鼠皮膚色素沉著的誘導(dǎo)作用,希望能夠?qū)ζ渲委煱遵帮L(fēng)的作用機(jī)制、劑量選擇及安全性有初步了解,為其更好的應(yīng)用于臨床而提供一定的參考。 方法以正常健康棕黃色豚鼠皮膚為實(shí)驗(yàn)?zāi)Ｐ?經(jīng)背部脫毛處理后,取4個(gè)相鄰近的區(qū)域依次作為對(duì)照組和低、中、高劑量組。根據(jù)最小紅斑量(MED值)并參考預(yù)實(shí)驗(yàn)結(jié)果將照射起始劑量分別定為0、90 mJ/cm2、180 mJ/cm2、270 mJ/cm2(即0、1MED、2MED、3MED),首次照射后未發(fā)生紅斑者下次照射劑量增加10-20%；發(fā)生紅斑但下次照射前可恢復(fù)者維持原劑量；出現(xiàn)有癥狀的紅斑、脫屑,則暫停照射致皮損緩解,且下次照射劑量減少10-20%。每周照射兩次,共照射6周,每周對(duì)實(shí)驗(yàn)動(dòng)物照射部位進(jìn)行觀察、拍攝,按時(shí)記錄照射區(qū)皮膚色素沉著情況。全部實(shí)驗(yàn)結(jié)束后處死動(dòng)物,分別取各組皮膚標(biāo)本進(jìn)行組織病理染色和免疫組織化學(xué)分析。實(shí)驗(yàn)分別采用肉眼評(píng)估、黑素細(xì)胞染色(Imokawa法)以及黑素顆粒染色(Masson-Fontana法)方法,研究中波高能紫外光所致色素沉著作用；應(yīng)用免疫組織化學(xué)染色法對(duì)不同照射劑量組豚鼠皮膚中的一氧化氮合酶(NOS)進(jìn)行檢測(cè),間接觀察黑素調(diào)節(jié)因子一氧化氮(NO)的變化情況；應(yīng)用組織病理常規(guī)染色方法(HE染色)觀察表皮經(jīng)中波高能紫外光照射后的組織學(xué)改變 結(jié)果全部豚鼠均完成實(shí)驗(yàn),少數(shù)豚鼠照射部位皮膚在照射初期出現(xiàn)紅斑、脫屑等不良反應(yīng),減少劑量或暫停照射后均緩解。隨著實(shí)驗(yàn)的進(jìn)行,豚鼠照射部位皮膚逐漸對(duì)中波高能紫外光產(chǎn)生耐受,不良反應(yīng)未再出現(xiàn)。各照射組均出現(xiàn)不同程度的色素沉著,除中、高劑量組色素沉著評(píng)分結(jié)果無(wú)顯著性差異外,其余各組的色素沉著評(píng)分、平均黑素?cái)?shù)量以及多巴陽(yáng)性黑素細(xì)胞數(shù)之間的差異均具有統(tǒng)計(jì)學(xué)意義(P0.05),且中劑量組結(jié)果優(yōu)于高劑量組；對(duì)照組無(wú)明顯改變。免疫織化學(xué)染色示iNOS的表達(dá)亦呈劑量依賴(lài)式遞增,各組間的差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。HE染色結(jié)果顯示各照射組表皮增生呈劑量依賴(lài)式遞增,與紫外線照射正常皮膚所致的組織形態(tài)學(xué)改變相符。未觀察到各劑量組中波高能紫外光照射引起的色素脫失及表皮細(xì)胞的異形性改變。 結(jié)論中波高能紫外光可誘導(dǎo)豚鼠皮膚色素沉著,而且這種效應(yīng)具有一定的劑量依賴(lài)性,但并非劑量越高越好。選擇接近或略高于最小紅斑量(MED)劑量作為初始劑量進(jìn)行照射,可有效減少累積劑量,在取得滿意療效的同時(shí)又可降低紫外線治療的潛在致癌風(fēng)險(xiǎn)。
[Abstract]:Objective to study the mechanism, dose selection and safety of the treatment of vitiligo by studying the inducing effect of high energy ultraviolet light on pigmentation in guinea pig skin. To provide a certain reference for its better application in clinical. Methods the normal brown guinea pig skin was used as the experimental model. After hair removal on the back, four adjacent areas were selected as control group and low, middle and high dose groups. According to the minimum erythema amount (MED) and referring to the results of pre-experiment, the initial dose of irradiation was determined to be 0 ~ 90 mJ/cm2180 mJ/cm2270 mJ/cm2 (i.e., 0 ~ 1 MED ~ (2) MED ~ (3 MED). The next irradiation dose of those without erythema was increased by 10 ~ (-20)% after the first irradiation. If erythema occurred but could recover before the next irradiation, the original dose could be maintained; if there were symptomatic erythema and detritus, the skin damage would be alleviated by pausing irradiation, and the dose of the next irradiation would be reduced by 10-20. The rats were irradiated twice a week for 6 weeks. The experimental animals were observed and photographed, and the pigmentation of the skin was recorded on time. At the end of the experiment, the animals were killed, and the skin samples of each group were stained with histopathology and analyzed by immunohistochemistry. The effects of medium wave high energy ultraviolet light on pigmentation were studied by means of naked eye evaluation, melanocyte staining (Imokawa) and melanin particle staining (Masson-Fontana). The nitric oxide synthase (NOS) in the skin of guinea pigs with different doses of irradiation was detected by immunohistochemical staining, and the changes of nitric oxide (NO) of melanin regulatory factor were observed indirectly. Histopathological routine staining (HE staining) was used to observe the histological changes of epidermis irradiated by medium wave high energy ultraviolet light in all guinea pigs. Erythema appeared in the skin of a few guinea pigs at the beginning of irradiation. Adverse reactions, such as detritus, were alleviated after reduction of dose or suspension of irradiation. With the development of the experiment, the skin of guinea pigs gradually developed tolerance to medium wave and high energy ultraviolet light, but the adverse reaction did not appear again. The pigmentation scores of each irradiation group were different, except that there was no significant difference in the pigmentation scores of the middle and high dose groups. The average melanin quantity and the number of dopa-positive melanocytes were significantly different (P0.05), and the results of middle dose group were better than that of high dose group. There was no obvious change in the control group. Immunocytochemical staining showed that the expression of iNOS was increased in a dose-dependent manner, and the differences among the groups were statistically significant (P0.05). The results of). HE staining showed that the epidermis proliferation of each irradiation group was increased in a dose-dependent manner. It is consistent with the histomorphologic changes caused by ultraviolet radiation on normal skin. The loss of pigment and the abnormal changes of epidermal cells were not observed in different dose groups. Conclusion Medium-wave high energy ultraviolet light can induce pigmentation in guinea pig skin in a dose-dependent manner, but the higher the dose is, the better. When the dose of (MED) is close to or slightly higher than the minimum amount of erythema as the initial dose, the cumulative dose can be effectively reduced, and the potential carcinogenic risk of UV treatment can be reduced while the curative effect is satisfactory.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R758.41
本文編號(hào)：2396161
[Abstract]:Objective to study the mechanism, dose selection and safety of the treatment of vitiligo by studying the inducing effect of high energy ultraviolet light on pigmentation in guinea pig skin. To provide a certain reference for its better application in clinical. Methods the normal brown guinea pig skin was used as the experimental model. After hair removal on the back, four adjacent areas were selected as control group and low, middle and high dose groups. According to the minimum erythema amount (MED) and referring to the results of pre-experiment, the initial dose of irradiation was determined to be 0 ~ 90 mJ/cm2180 mJ/cm2270 mJ/cm2 (i.e., 0 ~ 1 MED ~ (2) MED ~ (3 MED). The next irradiation dose of those without erythema was increased by 10 ~ (-20)% after the first irradiation. If erythema occurred but could recover before the next irradiation, the original dose could be maintained; if there were symptomatic erythema and detritus, the skin damage would be alleviated by pausing irradiation, and the dose of the next irradiation would be reduced by 10-20. The rats were irradiated twice a week for 6 weeks. The experimental animals were observed and photographed, and the pigmentation of the skin was recorded on time. At the end of the experiment, the animals were killed, and the skin samples of each group were stained with histopathology and analyzed by immunohistochemistry. The effects of medium wave high energy ultraviolet light on pigmentation were studied by means of naked eye evaluation, melanocyte staining (Imokawa) and melanin particle staining (Masson-Fontana). The nitric oxide synthase (NOS) in the skin of guinea pigs with different doses of irradiation was detected by immunohistochemical staining, and the changes of nitric oxide (NO) of melanin regulatory factor were observed indirectly. Histopathological routine staining (HE staining) was used to observe the histological changes of epidermis irradiated by medium wave high energy ultraviolet light in all guinea pigs. Erythema appeared in the skin of a few guinea pigs at the beginning of irradiation. Adverse reactions, such as detritus, were alleviated after reduction of dose or suspension of irradiation. With the development of the experiment, the skin of guinea pigs gradually developed tolerance to medium wave and high energy ultraviolet light, but the adverse reaction did not appear again. The pigmentation scores of each irradiation group were different, except that there was no significant difference in the pigmentation scores of the middle and high dose groups. The average melanin quantity and the number of dopa-positive melanocytes were significantly different (P0.05), and the results of middle dose group were better than that of high dose group. There was no obvious change in the control group. Immunocytochemical staining showed that the expression of iNOS was increased in a dose-dependent manner, and the differences among the groups were statistically significant (P0.05). The results of). HE staining showed that the epidermis proliferation of each irradiation group was increased in a dose-dependent manner. It is consistent with the histomorphologic changes caused by ultraviolet radiation on normal skin. The loss of pigment and the abnormal changes of epidermal cells were not observed in different dose groups. Conclusion Medium-wave high energy ultraviolet light can induce pigmentation in guinea pig skin in a dose-dependent manner, but the higher the dose is, the better. When the dose of (MED) is close to or slightly higher than the minimum amount of erythema as the initial dose, the cumulative dose can be effectively reduced, and the potential carcinogenic risk of UV treatment can be reduced while the curative effect is satisfactory.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R758.41
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