發(fā)布時間：2018-06-23 05:48

  本文選題：白癜風 + 紫外線�。� 參考：《山東大學》2011年博士論文

【摘要】：目的：評價定向中波高能紫外線(以下簡稱TH-UVB)與窄譜中波紫外線(NB-UVB)治療白癜風的療效與安全性。 方法：采用自身對照的方法,對33名白癜風患者進行觀察。取白癜風患者自身對稱或相鄰的皮損為靶皮損,隨機選擇試驗側和對照側分別使用TH-UVB或NB-UVB進行照射。試驗側治療前測定最小紅斑量(MED),初次照射選用2倍MED,根據(jù)前次治療反應決定下一次劑量。如前次治療后未出現(xiàn)清晰紅斑,下次劑量增加10%；出現(xiàn)持續(xù)清晰可見的紅斑后,每次維持原有劑量；如出現(xiàn)疼痛或水皰等,則暫停治療,癥狀消退后下次照射劑量減少10%。對照側初次劑量一般為70%的平均MED,根據(jù)前次治療反應決定下一次劑量。如前次治療后局部無紅斑或紅斑持續(xù)時間24h,下次劑量增加10%；紅斑持續(xù)24～72h,維持原照射劑量不變；如紅斑超過72h或出現(xiàn)水皰,則暫停治療,癥狀消退后下次照射劑量減少10%。每周治療2次,共治療12周。治療前及治療后每周拍照對比,記錄患者皮損處色素再生面積與治療前皮損面積的百分比,同時比較不同年齡、性別、發(fā)病年齡、病程、部位、臨床類型的療效。用SPSS統(tǒng)計軟件分析數(shù)據(jù),并記錄不良反應。 結果：3例患者未完成試驗,最終納入分析30例患者。其中男14例,女16例；年齡19～56歲,平均(34.3±8.9)歲。皮損位于面頸部5例,軀干部14例,四肢8例,手足3例。治療12周后,試驗側有效率為56.7%,對照側為20.0%,經(jīng)卡方檢驗,差異有統(tǒng)計學意義(P0.05)。試驗側開始色素再生的照射次數(shù)平均為(7.95±3.43)次。對照側平均為(15.36±3.43)次,經(jīng)配對樣本t檢驗,差異有統(tǒng)計學意義(P0.05)。經(jīng)Logistic回歸分析各臨床特征與臨床療效的相關性,顯示療效與患者發(fā)病部位、臨床類型、年齡、性別、發(fā)病年齡、病程等均無相關性(P均0.05)。試驗側不良反應發(fā)生率較對照側高(P0.05),但均輕微且不影響治療。 結論：TH-UVB與NB-UVB相比治療白癜風療效更好、起效更快,且安全性均較好。 目的：探討不同劑量TH-UVB與NB-UVB照射對豚鼠皮膚色素沉著及表達α-黑素細胞刺激素(α-MSH)、內(nèi)皮素-1(ET-1)的影響。 方法：以正常棕黃色豚鼠為實驗模型,在其背部取五塊相離的區(qū)域分為TH-UVB高、低劑量組、NB-UVB高、低劑量組及空白對照組進行照射。TH-UVB高、低劑量組起始劑量分別為270、90mJ/cm2,照射后發(fā)生持續(xù)24h以上紅斑即每次維持原有劑量,如果初次治療后未發(fā)生紅斑,則下次劑量增加10%,發(fā)生紅斑但24h內(nèi)消退者下次增加5%,如出現(xiàn)水皰、脫屑等,暫停治療至癥狀緩解,下次照射劑量減少10%。NB-UVB高、低劑量組起始劑量分別為200、100 mJ/cm2,每次劑量增加20%,出現(xiàn)輕微紅斑則增加10%,出現(xiàn)清晰紅斑暫停一次治療,出現(xiàn)水皰、疼痛等,則暫停治療,癥狀消退后下次照射劑量減少10%。每周照射2次,共照射8周。空白對照組不予處理。每2周對實驗區(qū)域進行拍攝,并按時記錄豚鼠皮膚色素沉著情況。全部實驗結束后處死動物。分別取空白對照組與各實驗組照射區(qū)域皮膚組織,采用肉眼評估及黑素顆粒染色(Fontana-Masson法)研究其致色素沉著作用；通過免疫組化法和逆轉錄-聚合酶鏈式反應(RT-PCR法)檢測豚鼠皮膚中的a-MSH、ET-1蛋白及mRNA表達情況。 結果：部分豚鼠皮膚照射后出現(xiàn)脫屑等不良反應,減少劑量或暫停照射即可緩解。各照射組均在照射1周后出現(xiàn)色素沉著,TH-UVB組較為均勻、清晰,NB-UVB組僅見散在色沉斑片�？瞻讓φ战M未見明顯變化。隨著照射時間和累積劑量的增加,各組色素逐漸加深,分布逐漸均勻。各組的色素沉著評分、黑素顆粒含量差異均有統(tǒng)計學意義(P均0.05),TH-UVB高、低劑量組之間差異無統(tǒng)計學意義(P0.05),但均高于NB-UVB組(P0.05),且NB-UVB高劑量組高于低劑量組(P0.05)；各組的α-MSH免疫組化計分差異有統(tǒng)計學意義(P0.05),TH-UVB組高于NB-UVB組(P0.05),但每種光源不同劑量組之間差異無統(tǒng)計學意義(P0.05)；各組的a-MSH mRNA表達水平差異有統(tǒng)計學意義(P0.05)TH-UVB高、低劑量組之間以及NB-UVB低劑量組與空白對照組之間差異無統(tǒng)計學意義(P0.05),余兩兩比較差異均有統(tǒng)計學意義(P均0.05)；各組的ET-1免疫組化結果差異有統(tǒng)計學意義(P0.05), NB-UVB高、低劑量組之間差異無統(tǒng)計學意義(P0.05),其余各組兩兩比較,差異均有統(tǒng)計學意義(P均0.05)；各組的ET-1mRNA表達水平差異有統(tǒng)計學意義(P0.05),NB-UVB高、低劑量組之間差異無統(tǒng)計學意義(P0.05),其余各組兩兩比較,差異均有統(tǒng)計學意義(P均0.05) 結論：TH-UVB在致豚鼠皮膚色素沉著作用上優(yōu)于NB-UVB;該兩種光源均可促進豚鼠表皮中α-MSH和ET-1蛋白及mRNA水平的表達,且TH-UVB作用更為顯著。 目的：1.探討不同劑量TH-UVB照射對人角質(zhì)形成細胞株(HaCaT細胞)a-MSH、ET-1蛋白水平及(?)nRNA水平的影響,并與NB-UVB進行比較,探討兩種光源的差別； 2.探討300mJ/cm2 TH-UVB照射后不同時間點HaCaT細胞a-MSH、ET-1蛋白水平與(?)nRNA水平的變化。 方法：常規(guī)培養(yǎng)HaCaT細胞,6孔培養(yǎng)板中每孔接種106個細胞。當細胞生長至80%融合時進行照射,細胞與光源間的照射距離為15cm。實驗設立兩組：劑量組分別用0、100、200、300、400、500 mJ/cm2的TH-UVB和NB-UVB照射HaCaT細胞,24h后收集上清液及細胞。ELISA方法測定培養(yǎng)上清液中a-MSH、ET-1分泌量,RT-PCR法測定細胞a-MSH、ET-1mRNA的表達量。時間組用300 mJ/cm2TH-UVB照射HaCaT細胞,分別在0、6、12、24、48、72h后用ELISA方法測定上清液中a-MSH、ET-1分泌量,RT-PCR法測定細胞a-MSH、ET-1mRNA的表達量。每組實驗重復3次。 結果：不同劑量的TH-UVB和NB-UVB都可以促進HaCaT細胞α-MSH的表達、分泌。TH-UVB在劑量為300～400mJ/cm2時,上清液中α-MSH濃度最高,明顯高于100～200mJ/cm2組(P均0.05)；細胞a-MSH mRNA表達量亦最大,在0～300mJ/cm2范圍內(nèi)隨照射劑量增加而增加。400mJ/cm2時,培養(yǎng)細胞出現(xiàn)明顯皺縮、圓化,懸浮,α-MSH表達、分泌水平下降,劑量增至500mJ/cm2時更為顯著。NB-UVB在劑量為500mJ/cm2時α-MSH濃度最高,細胞a-MSH mRNA表達量亦最大,200～300mJ/cm2范圍內(nèi)無差別(P均0.05)。兩種光源同一劑量對比,TH-UVB 100-400mJ/cm2照射后HaCaT細胞的α-MSH分泌量均高于同劑量NB-UVB組(P均0.05),500mJ/cm2組沒有差別(P0.05)；200～400mJ/cm2照射后細胞a-MSH mRNA表達量均高于同劑量NB-UVB組(P均0.05),而100、500mJ/cm2照射后沒有差別(P均0.05)。 TH-UVB在200～500mJ/cm2劑量照射后,上清液中ET-1濃度均高于未照射組(P均0.05),但該照射梯度內(nèi)ET-1分泌量無差異(P均0.05)。分子水平上200mJ/cm2劑量時ET-1 mRNA表達開始增高,300mJ/cm2時達到高峰,400mJ/cm2時開始下降,500mJ/cm2時與未照射組無差異(P0.05)NB-UVB 100～500mJ/cm2照射后上清液中ET-1濃度均高于未照射組(P均0.05),在該照射梯度內(nèi)ET-1分泌量無差異(P均0.05)。但分子水平上200-500mJ/cm2時ET-1 mRNA表達量高于未照射組且在該照射梯度內(nèi)呈劑量依賴(P均0.05),照射劑量達500mJ/cm2時ET-1 mRNA表達量最高。兩種光源同一劑量比較,TH-UVB500mJ/cm2照射后上清液中的ET-1濃度高于同劑量NB-UVB組(P0.05),100-400mJ/cm2照射后沒有差別(P均0.05)；200～500mJ/cm2照射后細胞ET-1mRNA表達量均高于同劑量NB-UVB組(P均0.05),100mJ/cm2照射后沒有差別(P0.05) 300 mJ/cm2 TH-UVB照射后6h HaCaT細胞上清液中α-MSH濃度開始增高,24h達到高峰并維持在這一水平至72h；細胞α-MSH mRNA表達量也于6h開始增高,24h達到高峰,持續(xù)至48h,72h開始下降。300 mJ/cm2 TH-UVB照射后6h上清液ET-1濃度及細胞中ET-1 mRNA表達量均開始增高,并于48h達到高峰,72h開始下降。 結論：1.不同劑量的TH-UVB和NB-UVB都可以促進HaCaT細胞α-MSH和ET-1蛋白及(?)nRNA水平的表達,該效應每種光源各自在一定范圍內(nèi)呈劑量依賴性,且TH-UVB效力更強。 2.300 mJ/cm2 TH-UVB照射對HaCaT細胞α-MSH和ET-1蛋白及(?)nRNA水平的促進作用在48～72h時最為明顯。
[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.
【學位授予單位】：山東大學
【學位級別】：博士
【學位授予年份】：2011
【分類號】：R758.41

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