基于黑素合成的维药驱虫斑鸠菊中活性成分紫铆素、紫铆花素抗白癜风的作用机制研究

Anthelmintica Vernonia of Uighur Medicine is widely used to treat vitiligo as common medicinal herbs of Xinjiang vitiligo is a kind disease of skin pigment damage, It' occurrence and development has relationship with melanin synthesis broken. Pre-experiment showed that the butin and butein were identified from active ingredient from anthelmintica Vernonia by the fingerprinting techniques, and serum pharmaceutical chemistry, and spectrum-efficient combination of technology, and serum pharmacological method, but the specific mechanisms and the role of targets is unclear. In the present project, focusing on targets on where butin and butein act on ,from the key points of the melanin synthesis, tyrosinase family proteins, epidermal melanocyte unit cell system, the signaling pathway, alpha-melanocyte cells of estrogen and related perspectives angle to study the targets of butin and butein act on vitiligo, to explore mechanism of anti-vitiligo by ELISA, PCR, the CFDA-SE fluorescent labeling method technology..the study could lay the foundation for butin and butein applied as anti-vitiligo medicine.

维药驱虫斑鸠菊是治疗白癜风的常用药材，白癜风是一种色素障碍性皮肤病，它的发生发展均与黑素合成相关。课题组前期采用指纹图谱技术、血清药物化学、谱效结合技术及血清药理学等方法，初步确定驱虫斑鸠菊抗白癜风活性成分为紫铆素和紫铆花素，但具体机制及作用靶点尚不清楚。本项目以驱虫斑鸠菊活性成分紫铆素、紫铆花素为研究对象，利用ELISA、PCR、CFDA-SE荧光标记法等技术，围绕影响黑素合成的关键点，从酪氨酸酶家族蛋白、表皮黑素单元细胞体系、信号通路、α-黑素细胞雌激素及相关信号通路等角度研究紫铆素、紫铆花素作用靶点，探索其抗白癜风机制，为将驱虫斑鸠菊中的紫铆素、紫铆花素作为防治白癜风的先导化合物开发利用提供依据。

本研究采用血清指纹图谱方法验证了驱虫斑鸠菊中紫铆素、紫铆花素吸收入血的特点。体外以人A375黑素瘤细胞、黑素细胞-角质细胞共培养、多巴胺诱导的细胞凋亡等模型，考察了紫铆素、紫铆花素对黑素含量、细胞增殖及酪氨酸酶活性的影响；采用流式细胞技术测定细胞线粒体膜电位及钙离子含量，RT-PCR和western blot 法检测了酪氨酸酶及酪氨酸酶相关蛋白1、2以及MITF的基因和蛋白的表达，结果表明紫铆素能够显著促进黑素细胞增殖，升高酪酸酶活性及黑素含量，能够加快细胞的增殖速度，调节细胞周期，升高细胞线粒体膜电位水平，降低细胞内钙离子含量，抑制细胞早期凋亡；同时紫铆素能够促进角质细胞分泌信号分子SCF和bFGF，间接促进黑素细胞增殖、产生黑素小体引起皮肤色素沉着，而紫铆花素作用不是很明显，但紫铆素和紫铆花素均能够对多巴胺诱导的细胞凋亡有一定的保护作用。信号通路研究结果表明紫铆素能够调节SCF/c-kit通路中靶点基因SCF、c-kit的mRNA和蛋白表达水平，促进黑素合成。体内考察了紫铆素和紫铆花素对氢醌诱导的白癜风小鼠模型的影响，结果表明紫铆素、紫铆花素均能改善氢醌诱导的白癜风小鼠的毛发颜色，促进黑色素毛囊的增多，且对动物酪氨酸酶和酪氨酸酶相关蛋白-1的表达有显著的促进作用。本项目的研究结果为今后开发治疗白癜风的新药具有很大的参考意义。