基于雌激素（ER）水平-水通道蛋白3（AQP3）表达的丹参对皮肤氧化损伤保护作用及其机制研究

AQP3 that functions in the skin aging process expresses highly in the skin. The level of estrogen plays an important role in the degree of skin aging. Estrogen up-regulates the expression of APQ3 to increase the water content and the penetration barrier function of the stratum corneunm, maintain the skin elasticity and prevent the oxidation damage to skin. Extravasated blood is the root cause of aging in the view of Traditional Chinese Medicine, and DanShen is the typical medicine for activating blood circulation among the Traditional Chinese herb-medicine. Danshen has the same function as estrogen. In our reach,we use the human breast cancer cell proliferation experience to test the key element in Danshen which has the same function as estrogen. We also use immunohistochemistry RT-PCR and mitochondria atpase detection to reach that Danshen functioning as estrogen up-regulates the expression of APQ3 to regulate the water content, the penetration barrier function and the oxidative stress, in order to protect the skin and delay the skin aging. Explaining the activating blood circulation function of the main target protein by the modern molecular medicine provides the pharmacological evidence for clinical evaluation and theoretical basis for clinical medicine of delaying the skin aging.

水通道蛋白3（AQP3）在皮肤上有高的表达，AQP3参与皮肤衰老过程。雌激素的水平对皮肤的衰老程度至关重要。雌激素通过上调AQP3的表达，增加角质层含水量、渗透屏障功能及对皮肤氧化损伤的保护作用。祖国医学认为瘀血是衰老的根本原因，丹参是中医活血化瘀法的代表性药物。丹参具有雌激素效应。 本实验应用人乳腺癌MCF7细胞增殖实验评价丹参植物雌激素活性的关键组分；应用角质形成细胞衰老模型和小鼠皮肤衰老模型，采用免疫组化、实时定量荧光RT一PCR检测，线粒体ATP酶检测等方法，研究丹参通过雌激素效应上调AQP3的表达，调节角质层含水量、渗透屏障功能及氧化应激等，以发挥对皮肤的保护作用，延缓皮肤的衰老。 从现代医学分子水平解释其"活血"效应的主要靶蛋白,可为临床评价"活血"效应与机制提供药理学依据。为延缓皮肤衰老的临床用药提供理论依据。

本研究应用人角质形成细胞（HaCaT）及UV照射光损伤小鼠模型，研究丹参促进创面损伤修复机制、丹参对中波紫外线（UVB）照射后HaCaT的保护作用及丹参对UVB和长波紫外线（UVA）照射致小鼠皮肤损伤的保护作用。研究发现丹参酮ⅡA能通过促进细胞内ROS释放激活ERK磷酸化信号通路，具有促HaCaT细胞迁移作用，可能通过促使HaCaT细胞释放低水平的ROS，激活Erk磷酸化，上调基质金属蛋白酶MMP-2、MMP-9降解Fibronectin和Collagen-Ⅰ，促进细胞迁移。丹参素能够通过减少UVB辐射后细胞内ROS的生成及抑制UVB辐射后细胞与凋亡相关蛋白表达、增加抗凋亡相关蛋白表达，从而抑制UVB对皮肤细胞的损伤，丹参能够提高光老化模型小鼠皮肤中谷胱甘肽过氧化物酶GSH-PX活力，以及GSH/GSSG，同时也可提高皮肤中的含水量发挥其抗光老化作用。上述研究为丹参在中医皮肤美容的临床应用提供了实验室依据。