【摘要】：目的:1.觀察亞慢性砷染毒小鼠皮膚組織形態(tài)的變化,并對黑色素細胞進行特殊染色,尋找砷致皮膚色素異常的病理依據(jù)。2.研究亞慢性砷染毒小鼠皮膚黑色素含量的變化,并用NAC進行干預,探討砷致皮膚色素異常的分子生物學機制,尋找砷中毒的生物學標志。方法:選取6周齡健康雌性C57BL/6J小鼠50只,隨機分為五組,每組10只,對照組自由飲自來水,實驗組分別自由飲含亞砷酸鈉50、500、5000μg/L的水溶液,干預組自由飲含亞砷酸鈉5000μg/L的砷水30天后,隔天灌胃NAC,劑量20mg/kg體重,繼續(xù)飲5000μg/L的砷水。染毒60天以后,頸動脈取血、剃毛取皮膚。分析比較小鼠各組的體重和臟器系數(shù)。取小鼠皮膚做HE染色和多巴染色。測定皮膚酪氨酸酶(TYR)、全血和皮膚的還原型谷胱甘肽(GSH)、全血和皮膚的半胱氨酸、皮膚褐黑素和優(yōu)黑素的含量。結果:1.小鼠的一般毒性各染毒組和干預組小鼠體重差異無統(tǒng)計學意義(P0.05);各染毒組和干預組小鼠心、肝、脾、肺、腎臟器系數(shù)差異無統(tǒng)計學意義(P0.05)。2.小鼠皮膚組織形態(tài)學變化HE染色未見炎性浸潤,各組毛囊腔內(nèi)黑素顆粒有差異;多巴染色陽性率分別為對照組71.43%,低劑量組66.67%,中劑量組為33.33%,高劑量組為33.33%,干預組為33.33%,總體差異有統(tǒng)計學意義(P0.05)。3.小鼠皮膚酪氨酸酶(TYR)含量隨著砷濃度的增加,對照、低、中、高劑量組小鼠皮膚TYR含量呈降低趨勢;與對照組相比,中、高劑量組和干預組皮膚TYR含量降低(P0.05);干預組和高劑量組相比,TYR含量差異無統(tǒng)計學差異(P0.05)。4.小鼠全血和皮膚半胱氨酸含量與對照組相比,低、中、高劑量組全血半胱氨酸含量降低(P0.05),低、中、高劑量組和干預組皮膚半胱氨酸含量降低(P0.05);與高劑量組相比,干預組全血和皮膚半胱氨酸含量均升高(P0.05)。5.小鼠全血和皮膚GSH含量與對照組相比,中、高劑量組全血及皮膚GSH含量均降低(P0.05);與高劑量組相比,干預組全血及皮膚GSH含量均升高(P0.05)。6.小鼠皮膚優(yōu)黑素和褐黑素的含量與對照組相比,低、中、高劑量組和干預組皮膚褐黑素含量降低(P0.05),中、高劑量組和干預組皮膚優(yōu)黑素和總黑素含量降低(P0.05),低、中、高劑量組和干預組褐黑素/優(yōu)黑素值降低(P0.05)。與高劑量組相比,干預組褐黑素、褐黑素/優(yōu)黑素值升高(P0.05)。結論:1.亞慢性砷暴露能夠使小鼠體內(nèi)TYR含量降低,TYR可能是砷暴露的早期生物學標志;2.亞慢性砷暴露能夠影響小鼠體內(nèi)非蛋白巰基的代謝,補充巰基能夠增加體內(nèi)非蛋白巰基的水平;3.亞慢性砷暴露能夠影響小鼠皮膚黑色素的含量,補充巰基能夠增加皮膚褐黑素的水平,巰基物質能夠在一定程度上緩解砷的皮膚毒性。
[Abstract]:Objective: 1. To observe the changes of skin tissue morphology in mice exposed to subchronic arsenic, and to make special staining on melanocytes to find the pathological basis of arsenic-induced skin pigmentation abnormality. 2. To study the changes of melanin content in skin of mice exposed to subchronic arsenic, to explore the molecular biological mechanism of arsenic-induced skin pigment abnormality and to search for the biological markers of arsenic poisoning. Methods: fifty 6-week-old healthy female C57BL/6J mice were randomly divided into five groups, 10 rats in each group. The control group drank tap water freely, and the experimental group drank water solution containing 500 渭 g / L sodium arsenite, 5 000 渭 g / L sodium arsenite, respectively. After 30 days of free drinking arsenic water containing 5000 渭 g / L sodium arsenite, the intervention group was given NAC, dose of 20mg/kg every other day and continued to drink 5000 渭 g / L arsenic water. After 60 days of exposure, carotid blood was taken and skin was shaved. The body weight and organ coefficient of each group were analyzed and compared. The skin of mice was stained with HE and dopa. The contents of cysteine, melanin and eumelanin in the whole blood of skin tyrosinase (TYR), and the whole blood of reduced glutathione (GSH), and in skin were determined. Results: 1. There was no significant difference in body weight between each group and intervention group (P0.05), while there was no significant difference in organ coefficient of heart, liver, spleen, lung and kidney between each group and intervention group (P0.05). HE staining showed no inflammatory infiltration, and melanin granules in hair follicles were different in each group. The positive rates of dopa staining were 71.43 in the control group, 66.67 in the low dose group, 33.33 in the middle dose group, 33.33 in the high dose group and 33.33 in the intervention group, respectively. The overall difference was statistically significant (P0.05). The content of tyrosinase (TYR) in mouse skin decreased with the increase of arsenic concentration in control, low, medium and high dose groups, and decreased in high dose group and intervention group compared with control group (P0.05). There was no significant difference in TYR content between the intervention group and the high dose group (P0.05). Compared with the control group, the content of cysteine in whole blood and skin of mice was lower than that in control group (P0.05), and the content of cysteine in the skin of middle, high and high dose groups was lower than that of control group (P0.05). Compared with the high dose group, the whole blood and skin cysteine content in the intervention group increased (P0.05). Compared with the control group, the content of GSH in whole blood and skin decreased in high dose group (P0.05), and the GSH content in whole blood and skin increased in intervention group compared with high dose group (P0.05). Compared with the control group, the content of skin eumelanin and melanin in mice was lower than that in the control group (P0.05), while the content of melanin and total melanin in the skin of high dose group and intervention group were lower than that of control group (P 0.05), and the content of melanin in skin of high dose group and intervention group was lower than that of control group (P0.05). The values of melanin / eumelanin in low, medium, high dose and intervention groups were decreased (P0.05). Compared with the high dose group, the values of melanin, melanin / eumelanin in the intervention group were higher than those in the intervention group (P0.05). Conclusion: 1. Subchronic arsenic exposure can reduce the TYR content in mice, TYR may be an early biological marker of arsenic exposure; 2. Subchronic arsenic exposure can affect the metabolism of non-protein sulfhydryl group in mice, and supplement sulfhydryl group can increase the level of non-protein sulfhydryl group in mice. Subchronic arsenic exposure can affect the content of melanin in skin of mice, and supplementation of sulfhydryl can increase the level of melanin in skin, and sulfhydryl can alleviate the skin toxicity of arsenic to a certain extent.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R114
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本文編號：2411908