定向中波高能紫外线治疗白癜风的临床和实验研究

发布时间：2018-06-23 05:48

本文选题：白癜风 + 紫外线　；参考：《山东大学》2011年博士论文

【摘要】：目的：评价定向中波高能紫外线(以下简称TH-UVB)与窄谱中波紫外线(NB-UVB)治疗白癜风的疗效与安全性。方法：采用自身对照的方法,对33名白癜风患者进行观察。取白癜风患者自身对称或相邻的皮损为靶皮损,随机选择试验侧和对照侧分别使用TH-UVB或NB-UVB进行照射。试验侧治疗前测定最小红斑量(MED),初次照射选用2倍MED,根据前次治疗反应决定下一次剂量。如前次治疗后未出现清晰红斑,下次剂量增加10%；出现持续清晰可见的红斑后,每次维持原有剂量；如出现疼痛或水疱等,则暂停治疗,症状消退后下次照射剂量减少10%。对照侧初次剂量一般为70%的平均MED,根据前次治疗反应决定下一次剂量。如前次治疗后局部无红斑或红斑持续时间24h,下次剂量增加10%；红斑持续24～72h,维持原照射剂量不变；如红斑超过72h或出现水疱,则暂停治疗,症状消退后下次照射剂量减少10%。每周治疗2次,共治疗12周。治疗前及治疗后每周拍照对比,记录患者皮损处色素再生面积与治疗前皮损面积的百分比,同时比较不同年龄、性别、发病年龄、病程、部位、临床类型的疗效。用SPSS统计软件分析数据,并记录不良反应。结果：3例患者未完成试验,最终纳入分析30例患者。其中男14例,女16例；年龄19～56岁,平均(34.3±8.9)岁。皮损位于面颈部5例,躯干部14例,四肢8例,手足3例。治疗12周后,试验侧有效率为56.7%,对照侧为20.0%,经卡方检验,差异有统计学意义(P0.05)。试验侧开始色素再生的照射次数平均为(7.95±3.43)次。对照侧平均为(15.36±3.43)次,经配对样本t检验,差异有统计学意义(P0.05)。经Logistic回归分析各临床特征与临床疗效的相关性,显示疗效与患者发病部位、临床类型、年龄、性别、发病年龄、病程等均无相关性(P均0.05)。试验侧不良反应发生率较对照侧高(P0.05),但均轻微且不影响治疗。结论：TH-UVB与NB-UVB相比治疗白癜风疗效更好、起效更快,且安全性均较好。目的：探讨不同剂量TH-UVB与NB-UVB照射对豚鼠皮肤色素沉着及表达α-黑素细胞刺激素(α-MSH)、内皮素-1(ET-1)的影响。方法：以正常棕黄色豚鼠为实验模型,在其背部取五块相离的区域分为TH-UVB高、低剂量组、NB-UVB高、低剂量组及空白对照组进行照射。TH-UVB高、低剂量组起始剂量分别为270、90mJ/cm2,照射后发生持续24h以上红斑即每次维持原有剂量,如果初次治疗后未发生红斑,则下次剂量增加10%,发生红斑但24h内消退者下次增加5%,如出现水疱、脱屑等,暂停治疗至症状缓解,下次照射剂量减少10%。NB-UVB高、低剂量组起始剂量分别为200、100 mJ/cm2,每次剂量增加20%,出现轻微红斑则增加10%,出现清晰红斑暂停一次治疗,出现水疱、疼痛等,则暂停治疗,症状消退后下次照射剂量减少10%。每周照射2次,共照射8周。空白对照组不予处理。每2周对实验区域进行拍摄,并按时记录豚鼠皮肤色素沉着情况。全部实验结束后处死动物。分别取空白对照组与各实验组照射区域皮肤组织,采用肉眼评估及黑素颗粒染色(Fontana-Masson法)研究其致色素沉着作用；通过免疫组化法和逆转录-聚合酶链式反应(RT-PCR法)检测豚鼠皮肤中的a-MSH、ET-1蛋白及mRNA表达情况。结果：部分豚鼠皮肤照射后出现脱屑等不良反应,减少剂量或暂停照射即可缓解。各照射组均在照射1周后出现色素沉着,TH-UVB组较为均匀、清晰,NB-UVB组仅见散在色沉斑片。空白对照组未见明显变化。随着照射时间和累积剂量的增加,各组色素逐渐加深,分布逐渐均匀。各组的色素沉着评分、黑素颗粒含量差异均有统计学意义(P均0.05),TH-UVB高、低剂量组之间差异无统计学意义(P0.05),但均高于NB-UVB组(P0.05),且NB-UVB高剂量组高于低剂量组(P0.05)；各组的α-MSH免疫组化计分差异有统计学意义(P0.05),TH-UVB组高于NB-UVB组(P0.05),但每种光源不同剂量组之间差异无统计学意义(P0.05)；各组的a-MSH mRNA表达水平差异有统计学意义(P0.05)TH-UVB高、低剂量组之间以及NB-UVB低剂量组与空白对照组之间差异无统计学意义(P0.05),余两两比较差异均有统计学意义(P均0.05)；各组的ET-1免疫组化结果差异有统计学意义(P0.05), NB-UVB高、低剂量组之间差异无统计学意义(P0.05),其余各组两两比较,差异均有统计学意义(P均0.05)；各组的ET-1mRNA表达水平差异有统计学意义(P0.05),NB-UVB高、低剂量组之间差异无统计学意义(P0.05),其余各组两两比较,差异均有统计学意义(P均0.05) 结论：TH-UVB在致豚鼠皮肤色素沉着作用上优于NB-UVB;该两种光源均可促进豚鼠表皮中α-MSH和ET-1蛋白及mRNA水平的表达,且TH-UVB作用更为显著。目的：1.探讨不同剂量TH-UVB照射对人角质形成细胞株(HaCaT细胞)a-MSH、ET-1蛋白水平及(?)nRNA水平的影响,并与NB-UVB进行比较,探讨两种光源的差别； 2.探讨300mJ/cm2 TH-UVB照射后不同时间点HaCaT细胞a-MSH、ET-1蛋白水平与(?)nRNA水平的变化。方法：常规培养HaCaT细胞,6孔培养板中每孔接种106个细胞。当细胞生长至80%融合时进行照射,细胞与光源间的照射距离为15cm。实验设立两组：剂量组分别用0、100、200、300、400、500 mJ/cm2的TH-UVB和NB-UVB照射HaCaT细胞,24h后收集上清液及细胞。ELISA方法测定培养上清液中a-MSH、ET-1分泌量,RT-PCR法测定细胞a-MSH、ET-1mRNA的表达量。时间组用300 mJ/cm2TH-UVB照射HaCaT细胞,分别在0、6、12、24、48、72h后用ELISA方法测定上清液中a-MSH、ET-1分泌量,RT-PCR法测定细胞a-MSH、ET-1mRNA的表达量。每组实验重复3次。结果：不同剂量的TH-UVB和NB-UVB都可以促进HaCaT细胞α-MSH的表达、分泌。TH-UVB在剂量为300～400mJ/cm2时,上清液中α-MSH浓度最高,明显高于100～200mJ/cm2组(P均0.05)；细胞a-MSH mRNA表达量亦最大,在0～300mJ/cm2范围内随照射剂量增加而增加。400mJ/cm2时,培养细胞出现明显皱缩、圆化,悬浮,α-MSH表达、分泌水平下降,剂量增至500mJ/cm2时更为显著。NB-UVB在剂量为500mJ/cm2时α-MSH浓度最高,细胞a-MSH mRNA表达量亦最大,200～300mJ/cm2范围内无差别(P均0.05)。两种光源同一剂量对比,TH-UVB 100-400mJ/cm2照射后HaCaT细胞的α-MSH分泌量均高于同剂量NB-UVB组(P均0.05),500mJ/cm2组没有差别(P0.05)；200～400mJ/cm2照射后细胞a-MSH mRNA表达量均高于同剂量NB-UVB组(P均0.05),而100、500mJ/cm2照射后没有差别(P均0.05)。 TH-UVB在200～500mJ/cm2剂量照射后,上清液中ET-1浓度均高于未照射组(P均0.05),但该照射梯度内ET-1分泌量无差异(P均0.05)。分子水平上200mJ/cm2剂量时ET-1 mRNA表达开始增高,300mJ/cm2时达到高峰,400mJ/cm2时开始下降,500mJ/cm2时与未照射组无差异(P0.05)NB-UVB 100～500mJ/cm2照射后上清液中ET-1浓度均高于未照射组(P均0.05),在该照射梯度内ET-1分泌量无差异(P均0.05)。但分子水平上200-500mJ/cm2时ET-1 mRNA表达量高于未照射组且在该照射梯度内呈剂量依赖(P均0.05),照射剂量达500mJ/cm2时ET-1 mRNA表达量最高。两种光源同一剂量比较,TH-UVB500mJ/cm2照射后上清液中的ET-1浓度高于同剂量NB-UVB组(P0.05),100-400mJ/cm2照射后没有差别(P均0.05)；200～500mJ/cm2照射后细胞ET-1mRNA表达量均高于同剂量NB-UVB组(P均0.05),100mJ/cm2照射后没有差别(P0.05) 300 mJ/cm2 TH-UVB照射后6h HaCaT细胞上清液中α-MSH浓度开始增高,24h达到高峰并维持在这一水平至72h；细胞α-MSH mRNA表达量也于6h开始增高,24h达到高峰,持续至48h,72h开始下降。300 mJ/cm2 TH-UVB照射后6h上清液ET-1浓度及细胞中ET-1 mRNA表达量均开始增高,并于48h达到高峰,72h开始下降。结论：1.不同剂量的TH-UVB和NB-UVB都可以促进HaCaT细胞α-MSH和ET-1蛋白及(?)nRNA水平的表达,该效应每种光源各自在一定范围内呈剂量依赖性,且TH-UVB效力更强。 2.300 mJ/cm2 TH-UVB照射对HaCaT细胞α-MSH和ET-1蛋白及(?)nRNA水平的促进作用在48～72h时最为明显。
[Abstract]:Objective: To evaluate the efficacy and safety of targeted medium wave high energy ultraviolet (TH-UVB) and narrow band ultraviolet B (NB-UVB) in the treatment of vitiligo.
Methods: 33 patients with vitiligo were observed by self control. The skin lesions of vitiligo patients with self symmetry or adjacent skin lesions were taken as the target lesions. The test side and the control side were randomly selected to use TH-UVB or NB-UVB. The minimum erythema volume (MED) was measured before the test side, and 2 times of MED were selected for the first irradiation, according to the previous treatment. The next dose should be determined. If no clear erythema occurred after the previous treatment, the next dose increased by 10%; after the emergence of the persistent and visible erythema, the original dose was maintained each time; if pain or blister appeared, the treatment was suspended, and the next irradiation dose decreased after the symptoms subsided and the first dose of the 10%. control side was generally 70% of the average, according to the previous time. The treatment reaction decided the next dose. If the previous treatment, the local no erythema or erythema duration was 24h, the next dose increased by 10%, the erythema lasted 24 to 72h, and the original irradiation dose remained unchanged. If the erythema was more than 72h or the blisters, the treatment was suspended and the next dose of 10%. was reduced by 2 times a week after the symptom subsiding, and the treatment was treated for 12 weeks. The percentage of pigment regeneration area and skin lesion area before and after treatment were recorded before and after treatment. At the same time, the effects of different age, sex, age, course, location, and clinical type were compared. The data were analyzed with SPSS software, and the adverse reaction was recorded.
Results: 3 patients had not completed the test, and finally included 30 cases, including 14 men and 16 women; the average age was 19~56 years old (34.3 + 8.9) years old. The lesions were in 5 cases in the face and neck, 14 in the body, 8 in the limbs and 3 in the hands and feet. After 3 weeks of treatment, the experimental side was 56.7% and the control side was 20%. The difference was statistically significant (P0.05). The average exposure times of pigment regeneration at the experimental side were (7.95 + 3.43) times (7.95 + 3.43). The average of the control side was (15.36 + 3.43) times. The difference was statistically significant (P0.05) by paired sample t test. The correlation between the clinical features and clinical efficacy was analyzed by Logistic regression, and the therapeutic effect and the site, age, sex, age and age of the patients were revealed. There was no correlation between the duration of the disease (P = 0.05). The incidence of adverse reactions on the test side was higher than that in the control side (P0.05), but all of them were mild and did not affect the treatment.
Conclusion: compared with NB-UVB, TH-UVB has better efficacy, faster onset and better safety in the treatment of vitiligo.
Objective: To investigate the effects of different doses of TH-UVB and NB-UVB on pigmentation and expression of alpha melanocyte stimulating hormone (-MSH) and endothelin -1 (ET-1) in guinea pigs.
Methods: the normal brown yellow guinea pig was used as the experimental model. Five separate areas were divided into TH-UVB high, low dose group, NB-UVB high, low dose group and blank control group. The initial dose of low dose group was 270,90mJ/cm2, and the red spot above 24h after irradiation was maintained at the original dose, if the initial dose was maintained, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose was maintained for the first time, if the initial dose of TH-UVB was maintained, if the initial dose was maintained, the initial dose of the group was maintained at the same time, if the initial dose was maintained for the first time. After treatment, no erythema occurred, the next dose increased by 10%, and erythema was increased by 5% at the next 24h decline, such as blisters, desquamation, pause treatment to symptomatic relief, the next irradiation dose reduced by 10%.NB-UVB, the initial dose of the low dose group was 200100 mJ/cm2, each dose increased by 20%, and the occurrence of mild erythema was increased by 10%. The reddish erythema was suspended for one treatment, there were blisters and pain, and the treatment was suspended. After the symptom subsided, the next irradiation dose decreased by 10%. 2 times a week for 8 weeks. The blank control group was not treated. The experimental area was taken every 2 weeks and the pigmentation in the skin of the guinea pig was recorded on time. The animals were killed after the whole experiment. The animals were taken blank after the complete experiment. The skin tissue in the irradiated area of the control group and the experimental group was evaluated by the naked eye and melanin particle staining (Fontana-Masson method). The expression of a-MSH and ET-1 egg Rhizoma Bletillae mRNA in the skin of the guinea pig was detected by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR).
Results: some adverse reactions such as desquamation were found in the skin of some guinea pigs, reducing dose or pause irradiation could be alleviated. All groups of irradiated groups appeared pigmentation after 1 weeks of irradiation. Group TH-UVB was more uniform and clear. Group NB-UVB was only scattered in color sunk patches. The pigments of the group gradually deepened and distributed gradually. The difference of melanin content in each group was statistically significant (P 0.05), TH-UVB was high, and there was no significant difference between the low dose groups (P0.05), but higher than the group NB-UVB (P0.05), and the NB-UVB high dose group was higher than the low dose group (P0.05); the alpha -MSH immunization score difference of each group was different. The difference was statistically significant (P0.05), the group TH-UVB was higher than the NB-UVB group (P0.05), but there was no significant difference between the different doses of each light source (P0.05); the difference of a-MSH mRNA expression level in each group was statistically significant (P0.05) TH-UVB, and there was no statistical difference between the low dose group and the NB-UVB low dose group and the blank control group (P0.05) There were statistically significant differences between the two groups (P 0.05). The difference of ET-1 immunohistochemical results in each group was statistically significant (P0.05), NB-UVB was high, and there was no significant difference between the low dose groups (P0.05), the other 22 groups were statistically significant (P 0.05), and the difference of ET-1mRNA expression levels in each group was statistically significant. (P0.05) there was no significant difference between the NB-UVB high and low dose groups (P0.05), and the difference between the other 22 groups was statistically significant (P = 0.05).
Conclusion: TH-UVB is superior to NB-UVB in pigmentation of guinea pig skin, and these two light sources can promote the expression of alpha -MSH and ET-1 protein and mRNA in the epidermis of guinea pig, and the effect of TH-UVB is more significant.
Objective: 1. to investigate the effects of different doses of TH-UVB irradiation on the level of a-MSH, ET-1 protein and (?) nRNA in human keratinocyte (HaCaT cells), and to compare with NB-UVB to explore the difference between the two kinds of light sources.
2. to investigate the changes of a-MSH, ET-1 protein level and nRNA level in HaCaT cells at different time points after 300mJ/cm2 TH-UVB irradiation.
Methods: HaCaT cells were routinely cultured. 106 cells were inoculated each hole in the 6 Hole culture plate. When the cells grew to 80% fusion, the irradiation distance between the cells and the light source was set up in the 15cm. experiment. The dose group irradiated the HaCaT cells with 0100200300400500 mJ/cm2 TH-UVB and NB-UVB respectively, and then the supernatant and cell.ELISA were collected after 24h. Methods the secretion of a-MSH and ET-1 in the culture supernatant was measured. The expression of a-MSH and ET-1mRNA in cell was measured by RT-PCR. The time group irradiated HaCaT cells with 300 mJ/cm2TH-UVB, and the a-MSH, ET-1 secretion in the supernatant was measured by ELISA method after 0,6,12,24,48,72h, and the RT-PCR method was used to determine the expression amount. Each group repeated 3 times.
Results: different doses of TH-UVB and NB-UVB could promote the expression of alpha -MSH in HaCaT cells. When the secretion of.TH-UVB was 300 to 400mJ/cm2, the concentration of alpha -MSH in the supernatant was highest, obviously higher than that of the 100 ~ 200mJ/cm2 group (P 0.05), and the expression of a-MSH mRNA was also the largest, and increased with the increase of radiation dose in the 0 ~ 300mJ/cm2 range. At the time, the cultured cells showed obvious shrinkage, roundness, suspension, alpha -MSH expression and secretory level. When the dose increased to 500mJ/cm2, the concentration of alpha -MSH was the highest when the dose of 500mJ/cm2 was 500mJ/cm2, and the expression of a-MSH mRNA in the cell was also the largest. There was no difference in the range of 200 ~ 300mJ/cm2 (P 0.05). Two kinds of light sources were compared, TH-UVB 100-400mJ/cm2. The secretion of alpha -MSH in HaCaT cells after irradiation was higher than that in the same dose NB-UVB group (P 0.05), and there was no difference in 500mJ/cm2 group (P0.05). The mRNA expression of a-MSH in the cells after 200 ~ 400mJ/cm2 irradiation was higher than that in the same dose NB-UVB group (P 0.05), but there was no difference (0.05) after exposure to 100500mJ/cm2.
The concentration of ET-1 in the supernatant was higher than that in the unirradiated group (P 0.05) after 200 ~ 500mJ/cm2 dose irradiation, but there was no difference in ET-1 secretion in the irradiation gradient (P 0.05). At the molecular level, the mRNA expression of ET-1 began to increase, the 300mJ/cm2 reached the peak, and the 400mJ /cm2 began to decline, and there was no difference between the ET-1 and the unirradiated group. 05) the concentration of ET-1 in the supernatant after NB-UVB 100 ~ 500mJ/cm2 was higher than that in the unirradiated group (P 0.05), and there was no difference in ET-1 secretion in the irradiation gradient (P 0.05). But at the molecular level, the mRNA expression of ET-1 mRNA was higher than that in the unirradiated group, and the dosage of the ET-1 was dependent on the irradiation gradient (P 0.05), and the irradiation dose reached 500mJ/cm2 ET-1 Compared with the same dose of the two light sources, the concentration of ET-1 in the supernatant after TH-UVB500mJ/cm2 irradiation was higher than that in the same dose NB-UVB group (P0.05), and there was no difference after 100-400mJ/cm2 irradiation (P 0.05). The expression of ET-1mRNA in the cells after 200 ~ 500mJ/cm2 irradiation was higher than that in the same dose NB-UVB group (P 0.05), and there was no difference after 100mJ/cm2 irradiation (P0.05).
After 300 mJ/cm2 TH-UVB irradiation, the concentration of alpha -MSH in the supernatant of 6h HaCaT cells began to increase, 24h reached its peak and maintained at this level to 72h, and the mRNA expression of alpha -MSH increased at the beginning of 6h, and the 24h reached the peak. It began to increase and reached its peak at 48h, and 72h began to decline.
Conclusion: 1. different doses of TH-UVB and NB-UVB can promote the expression of alpha -MSH and ET-1 protein and (?) nRNA level in HaCaT cells. Each light source is dose-dependent in a certain range, and the potency of TH-UVB is stronger.
The promotion effect of 2.300 mJ/cm2 TH-UVB irradiation on the level of alpha -MSH and ET-1 protein and nRNA in HaCaT cells was most obvious at 48 ~ 72h.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R758.41

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