砷对小鼠皮肤黑色素合成的影响及NAC干预的研究

发布时间：2019-01-20 09:29

【摘要】：目的:1.观察亚慢性砷染毒小鼠皮肤组织形态的变化,并对黑色素细胞进行特殊染色,寻找砷致皮肤色素异常的病理依据。2.研究亚慢性砷染毒小鼠皮肤黑色素含量的变化,并用NAC进行干预,探讨砷致皮肤色素异常的分子生物学机制,寻找砷中毒的生物学标志。方法:选取6周龄健康雌性C57BL/6J小鼠50只,随机分为五组,每组10只,对照组自由饮自来水,实验组分别自由饮含亚砷酸钠50、500、5000μg/L的水溶液,干预组自由饮含亚砷酸钠5000μg/L的砷水30天后,隔天灌胃NAC,剂量20mg/kg体重,继续饮5000μg/L的砷水。染毒60天以后,颈动脉取血、剃毛取皮肤。分析比较小鼠各组的体重和脏器系数。取小鼠皮肤做HE染色和多巴染色。测定皮肤酪氨酸酶(TYR)、全血和皮肤的还原型谷胱甘肽(GSH)、全血和皮肤的半胱氨酸、皮肤褐黑素和优黑素的含量。结果:1.小鼠的一般毒性各染毒组和干预组小鼠体重差异无统计学意义(P0.05);各染毒组和干预组小鼠心、肝、脾、肺、肾脏器系数差异无统计学意义(P0.05)。2.小鼠皮肤组织形态学变化HE染色未见炎性浸润,各组毛囊腔内黑素颗粒有差异;多巴染色阳性率分别为对照组71.43%,低剂量组66.67%,中剂量组为33.33%,高剂量组为33.33%,干预组为33.33%,总体差异有统计学意义(P0.05)。3.小鼠皮肤酪氨酸酶(TYR)含量随着砷浓度的增加,对照、低、中、高剂量组小鼠皮肤TYR含量呈降低趋势;与对照组相比,中、高剂量组和干预组皮肤TYR含量降低(P0.05);干预组和高剂量组相比,TYR含量差异无统计学差异(P0.05)。4.小鼠全血和皮肤半胱氨酸含量与对照组相比,低、中、高剂量组全血半胱氨酸含量降低(P0.05),低、中、高剂量组和干预组皮肤半胱氨酸含量降低(P0.05);与高剂量组相比,干预组全血和皮肤半胱氨酸含量均升高(P0.05)。5.小鼠全血和皮肤GSH含量与对照组相比,中、高剂量组全血及皮肤GSH含量均降低(P0.05);与高剂量组相比,干预组全血及皮肤GSH含量均升高(P0.05)。6.小鼠皮肤优黑素和褐黑素的含量与对照组相比,低、中、高剂量组和干预组皮肤褐黑素含量降低(P0.05),中、高剂量组和干预组皮肤优黑素和总黑素含量降低(P0.05),低、中、高剂量组和干预组褐黑素/优黑素值降低(P0.05)。与高剂量组相比,干预组褐黑素、褐黑素/优黑素值升高(P0.05)。结论:1.亚慢性砷暴露能够使小鼠体内TYR含量降低,TYR可能是砷暴露的早期生物学标志;2.亚慢性砷暴露能够影响小鼠体内非蛋白巯基的代谢,补充巯基能够增加体内非蛋白巯基的水平;3.亚慢性砷暴露能够影响小鼠皮肤黑色素的含量,补充巯基能够增加皮肤褐黑素的水平,巯基物质能够在一定程度上缓解砷的皮肤毒性。
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R114
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本文编号：2411908