肉苁蓉有效成分调节色素代谢作用及其机制研究

Pigment metabolism disorders is a kind of common skin diseases, which is easy to diagnose but difficult to cure. It can be very harmful to patients, both physically and mentally. Even though there are many researches in the field of pigment metabolism disorders, it has lacked safe and effective drug in the treatment of pigment metabolism disorders. So developing new medicine from natural products has become the hot issue. The results from our preliminary studies suggested that the extract of phenylethanoid glycosides from cistanche was effective in treating hyperpigmentation. Our follow-up research showed phenylethanoid glycosides monomer components were capable of inhibiting pigmentation, but their activities were significant difference. On the basis of precedent researches, this project will adopt pharmacological experimental methods and molecular biological techniques to study effects of phenylethanoid glycosides monomer components on the biological actions, transcription and protein expression levels of related genes in melanocytes from pigmentation skin caused by ultraviolet radiation, and from normal skin of healthy persons are taken as controls in-vitro. And appearance of skin, histopathological manifestations, and melanin value of guinea-pig when exposed to ultraviolet radiation will be analyzed. Depended on the above studies and experiments, we hope to clarify the regulation mechanism of phenylethanoid glycosides to melanin metabolism. The structure activity relationship of phenylethanoid glycosides will also be studied. In brief, we aim to develop new drugs based on phenylethanoid glycosides against disturbance of pigment metabolism and provide a theoretical basis though the studies in the project.

色素代谢障碍性疾病是临床常见皮肤顽疾，易诊难治，给患者身体和心理造成严重伤害。目前临床尚缺乏有效的色素代谢障碍治疗药物，从天然产物发掘新的药物已成必然。课题组前期研究显示，肉苁蓉苯乙醇总苷提取物具有良好抑制皮肤色素沉着作用；后续研究又对从肉苁蓉中制备的系列苯乙醇苷单体进行了活性初筛，发现它们均具有一定的抑制色素沉着作用，但活性差异明显。本项目基于前期工作积累，拟采用现代药理学和分子生物学技术，于离体实验研究苯乙醇苷单体对正常人皮黑素细胞和紫外照射致色素沉着人皮黑素细胞的生物学作用及相关基因转录水平、蛋白表达情况；于在体实验研究其对紫外照射致豚鼠色素沉着模型皮肤外观反应、组织病理改变、黑素值影响等。综合分析，以阐明苯乙醇苷衍生物调节色素代谢作用机制。同时还将对其构效关系进行考察。通过本项目研究，力求发现抗色素代谢障碍新药，并为开发以苯乙醇苷为母核结构的新型抗色素代谢障碍药物提供理论依据。

本项目围绕肉苁蓉苯乙醇苷的抗皮肤色素沉着作用及机制展开了深入研究。在系统文献调研和大量预实验的基础上，不断优化完善研究方案，在项目实施的4年期间，按照原定计划：（1）建立了以人皮肤黑色素细胞为模型载体的黑素沉着离体模型，于细胞水平、分子层面研究了肉苁蓉苯乙醇总苷对正常人皮黑色素细胞的生物学作用（细胞活力、酪氨酸酶活性、黑素含量、细胞周期和细胞凋亡等），并考察其对黑素合成相关基因转录水平和蛋白表达情况的影响，深入探讨了其抗色素沉着的作用机制；（2）考察了10种苯乙醇苷单体化学成分对人皮黑色素细胞黑素合成的影响，分析了苯乙醇苷化学结构与抑制黑素合成作用之间的潜在关联，推断了苯乙醇苷类化学成分抗色素沉着的构效关系；（3）以获得高效快捷的皮肤外用给药系统为出发点，优选制备了以肉苁蓉苯乙醇苷为主要功效成分的纳米乳凝胶复合制剂，并对该制剂的质量控制和体外透皮吸收性能进行了研究；（4）在成功制备以肉苁蓉苯乙醇苷纳米乳凝胶复合制剂的基础上，构建了UVB紫外线诱导棕色豚鼠色素沉着动物模型，采用外观评价、激光共聚焦扫描成像、皮肤组织病理检测等方法对肉苁蓉苯乙醇苷抗色素代谢障碍的作用进行了全面综合的药效学评价。