摘要
酪氨酸酶(EC.1.14.18.1)是一种广泛存在于自然界中含铜的金属氧化酶,它具有单酚酶活性,可以将酪氨酸羟化形成L-多巴(L-DOPA),也具有二酚酶活力,将L-多巴氧化成多巴醌。它是生物形成黑色素合成的关键酶,决定了皮肤和头发色素的颜色。酪氨酸酶的异常表达引起了各种各样的皮肤病,如老年斑及光照损伤等。它还与人脑中的神经黑色素形成及神经退行性病变相关的帕金森病有关,因此酪氨酸酶抑制剂临床应用于与黑色素失调相关疾病的治疗。其在美白化妆品及日晒变黑应用方面具有重要作用。
本论文从天然植物中分离纯化出对酪氨酸酶具有很强抑制作用的抑制剂并探讨其对单酚酶和二酚酶活性的抑制作用机理。再以小鼠B16F10细胞的酪氨酸酶为研究对象,研究其对黑色素生成量的影响及其相关基因的转录调控机制。最后,以果蝇为动物模型研究酪氨酸酶抑制剂对酪氨酸酶代谢途径中相关产物多巴胺生成量的影响。在筛选酪氨酸酶抑制剂过程中,发现了与帕金森病相关的α-synuclein蛋白的过表达能显著抑制黑色素的生成量,这预示着酪氨酸酶与帕金森病可能有着某种相关性,因此研究了α-synuclein蛋白对小鼠B16F10细胞酪氨酸酶转录调控的机制。相关研究内容和结果如下:
(1)对酪氨酸酶有抑制作用的化合物:Cardol trienes,Cardol diene,Cardnol trienes,Cardanol diene,Cardanol monone,研究了它们对对酪氨酸酶的抑制机理及对小鼠B16F10细胞的黑色素生成量的影响及其相关基因的转录调控机制。
(2)从抗生素药品中筛选了对酪氨酸酶有制作用的药物,研究其对酪氨酸酶的抑制机理。
(3)以果蝇为动物模型研究酪氨酸酶抑制剂对酪氨酸酶代谢途径中相关物质含量的影响。
(4)研究了α-synuclein蛋白过表达对小鼠B16F10细胞的酪氨酸酶转录调控的机制。
Tyrosinase (1.14.18.1),widely distributed in nature,is a metalloenzyme oxidasewhich catalyzes two distinct reactions of melanin synthesis---the hydroxylation ofmonophenol and the oxidation of o-diphenol to the corresponding o-quinone.It is akey enzyme in melanin biosynthesis,involved in determining the color of mammalianskin and hair.Its abnormal expression is responsible for the various dermatologicaldisorders,such as melasama age spots,actinic damages.It also contributes toneuromelanin formation in the human brain and the neurodegeneration associatedwith Parkinson's disease.Tyrosinase inhibitors,therefore,should be clinically usefulfor the treatments of some dermatological disorders associated with melaninhyperpigmentation and also important in cosmetics for whitening and depigmentationafter sunburn.The inhibition of tyrosinase has been the subject of many studies.
In the present paper,we Separate inibitors from natural plants and effects of themon the activity of mushroom tyrosniase have been studied.Then effects of them on themelanin content and regulation of melanogenesis have been studied.In the end,effects of inhibitors on dopamine content in fly were assay.In the process of ourscreening the tyrosinase inhibitors,we found that melanin content could be depressionby overexpressingα-synuclein in the cell.The result indicates that tyrosinase andParkinson's disease have correlation to some extent,so the regulation of tyrosinase inthe B 16F10 has been studied.The contents and results were as follows:
Five compounds were seperated that can inhibit tyrosinase from CNSL:Cardoltrienes,Cardol diene,Cardnol trienes,Cardanol diene,Cardanol monone.Theinhibition mechanism of them against the activity of mushroom tyrosinase wasstudied.Effects of them on the melanin content and regulation of melanogenesis inthe B 16F 10 cell were also studied.
Screening the tyrosinase inhibitors from drugs and studying their inhibitorymechanism have been conducted.
Effects of inhibitors on dopamine content in fly were assay.Regulation ofmelanogenesis by overexpressingα-synuclein in the B 16F 10 was also studied.
引文
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