与结构及转运蛋白关联的川芎调控苷类成分血脑屏障转运机制的研究

Due to limited penetration of the blood-brain barrier (BBB) and extrusion of efflux protein, it was very difficult for many therapeutic agents for central nervous system（CNS） diseases in clinical to reach ideal curative effects. Research on a new and more efficient target vector was a trend of drug delivery system of CNS diseases. It was reported that not only the active components of Rhizoma Chuanxiong distributed widely in the body, but also they guided other to the focus of a disease. Though there was a little reports about the penetration of BBB of Rhizoma Chuanxiong, reports about the mechanism was little. And no research about which ingredients enhance the penetration of BBB was reported. Glycosides from herb were effective to CNS diseases, but were seldom used in clinical due to its poor availability or low penetration to BBB. Literatures and our researches showed that Rhizoma Chuanxiong could enhance the concentration of some glycosides, for example, Gastrodingenin, helicid, geniposide and jujuboside A, and decrease their elimination in brain. Based on the above results, we will carry on the following researches. Some glycosides which were effective to CNS diseases will be the model drug. Quantum chemistry, Circular dichroism spectra and BBB cell model will be employed for researches of interaction of drug-drug, drug-transfer protein and the further mechanism. All the researches will reveal which ingredients of Rhizoma Chuanxiong enhance the penetration of BBB, the mechanism of promote penetration, the relationship between penetration and structure, which will be a good method to find new potential specific carrier and promoter for drug delivery system of CNS diseases.

神经变性疾病（CNS疾病）药物研发瓶颈是血脑屏障（BBB），寻找促药物透BBB新方法是脑部给药系统研究重要方向。川芎有"上达颠顶，下至血海"和"引经"之功，其透BBB性能的报道较多，但川芎促其他药透BBB报道甚少，促透机制研究不够深入，未见川芎促药物透BBB物质基础研究。中药苷类成分对CNS疾病的治疗有重要作用，因透生物膜性能差，生物利用度低，限制其应用。文献报道，川芎常与含苷类成分中药配伍治疗CNS疾病，课题组发现川芎可提高一些苷类成分在脑组织浓度，延缓在脑内的清除。因此，本项目以治疗CNS疾病的中药苷类为模型药，运用量子化学、圆二色谱、BBB细胞模型等方法研究川芎活性成分与模型药、转运蛋白的相互作用和促透BBB能力，并进行促透作用与结构关联性分析，旨在阐明川芎促药物透BBB物质基础、作用机制和结构选择性规律，进一步寻找选择性BBB促透剂和药物载体，为解决药物透BBB难题提供新思路。

中枢神经（CNS）疾病药物研发瓶颈是血脑屏障（BBB），寻找促药物透BBB新方法是脑部给药系统研究重要方向。项目立足于川芎“上达颠顶，下至血海”和“引经”作用，以对CNS疾病有效、难以透过BBB的中药苷类成分为模型药，研究了川芎活性成分对模型药的促透作用、机制和物质基础。采用ADMET Predictor对川芎40个成分和天麻素、芍药苷、栀子苷、松果菊苷进行了透血脑屏障预测，结果川芎有30个成分为高渗透性，10个成分为低渗透性，其中阿魏酸属低渗透性，川芎嗪、洋川芎内酯A、洋川芎内酯I和藁本内酯属高渗透性，4种苷类模型药均为BBB低渗性。体外BBB细胞模型研究发现4种苷类成分均难以透过BBB，其转运存在旁路转运，并可能受到P-gp或MRP1外排蛋白影响。川芎活性成分能够提高苷类成分透BBB转运量，不同川芎成分复配有利提高苷类模型药的跨膜转运。Western blot法研究发现川芎中5个活性成分均能抑制P-gp蛋白表达和外排作用，并对MDR1 mRNA表达有抑制作用，川芎中洋川芎内酯A可能通过抑制MRP1表达来促进药物跨膜转运。当川芎5个活性成分达到一定浓度，它们可以抑制细胞膜上紧密连接蛋白ZO-1的表达，从而提高药物渗透性。Discovery Studio分子模拟结果显示，芍药苷、松果菊苷的LibDock Socre接近或大于阳性药维拉帕米，说明它们为P-gp底物，而天麻素、芍药苷以及松果菊苷的LibDock Socre接近或大于阳性药丙磺舒，说明这3个苷类成分为MRP1底物。川芎活性成分能调控细胞膜上Ca2+流动，能通过调控Na+-K+-ATP酶活性来调控细胞的渗透性。川芎活性成分能够下调脑组织中5-HT含量，这与其调控BBB通透性作用之间的关系，有待进一步研究。动物实验表明，川芎活性成分能够改变天麻素体内过程，并提高天麻活性成分在脑内的浓度，延缓其在脑组织中的清除。项目从体外细胞模型、整体动物、计算机软件预测等多方面阐明了川芎活性成分对苷类活性成分透BBB的作用、机制和物质基础，为解决药物难以透过BBB的瓶颈问题提供了新方法；培养了博士研究生1名，硕士研究生2名；发表论文6篇，其中SCI 3篇，中文核心期刊3篇。