南药“青天葵”抗肺纤维化药效物质及作用机制研究

Recently, the incidence and mortality of pulmonary fibrosis in whole world continue to increase, and there are no effective drugs until now. It is generally accepted that TGF-β1(Transforming growth factor β1) was the most closely related to the occurrence and proliferation of pulmonary fibrosis by activating TGF-β1 signal pathway through binding target cells. So blockade of the TGF-β1 signaling pathway as a novel therapeutic target may be a new choice for the treatment of these kinds of diseases. Herba Nerviliae,also called "Qing Tian Kui" in Southern China, is widely used for treatment of pulmonary-injury diseases. Previous chemical studies on this plant led to the isolation of ?avonoids and pharmacological survey showed that it can obviously improve the effect of anti-pulmonary fibrosis in rats. Its mechanism was refered to multi-targets with TGF-β1 signal transduction. Therefore,based on the theory as "multiple components with multiple targets or pathways for clinical efficiency" for Chinese medicine, guiding with anti PF activity, the active components of NF is prepared and clarified by LC/MS, etc.to reveal its active bases. Meanwhile, the systematic studies of active components on TGF-β1 signaling pathway as Smads and ERK1/2 will be performed to reveal its mechanism for anti-pulmonary fibrosis.

肺纤维化疾病发病率和死亡率持续上升，至今无有效治疗药。TGF-β1是公认的与肺纤维化发生和形成关系最为密切的介导因子，其通过受体介导，激活胞内信号分子蛋白转导信号，调控细胞胶原等效应基因的表达。抑制/阻断TGF-β1信号转导途径，可为肺纤维化治疗提供新手段。南药"青天葵"为岭南地区肺损伤疾病常用治疗药，前期研究证实其总黄酮具有抗肺纤维化作用，作用机制涉及TGF-β1信号通路的多个环节和靶点。本项目基于中药"多成分、多靶点、多途径发挥整体作用"的思路，以抗纤维化活性为导向，制备青天葵抗肺纤维化有效部位，解析有效部位化学组成，阐明药效物质并发现先导化合物；以有效部位对与肺纤维化发生发展密切相关的TGF-β1信号通路 (Smads通路和ERK1/ERK2通路)激活途径、关键信号分子的表达及调控、通路间的相互作用等的系统研究，揭示其抗肺纤维化作用的分子机制。

本项目综合运用现代分离与鉴定技术对南药“青天葵”中的化学成分进行分离、纯化、富集、鉴定，完成了南药“青天葵”中的化学成分系统研究，共分离得到35个化合物；从HP-20、AB-8、NKA-9、X-5、HPD-100和D101等六种大孔树脂中优选AB型大孔树脂进行工艺优化，制备青天葵总黄酮部位，以芦丁计总黄酮含量不低于20%。同时采用LC/MS/MS等技术，结合成分分离及结构鉴定研究基础，对有效部位组成成分进行解析、辨认，明确有效部位的物质基础；采用盐酸博莱霉素复制肺纤维化模型，对青天葵总黄酮部位及非黄酮部位进行药效学评价，结果表明，青天葵总黄酮部位和非黄酮部位均对盐酸博莱酶素诱导的肺纤维化大鼠具有保护作用，且以总黄酮部位效果为优，并阐明了其作用机制与调控TGF-β/Smad和TGF-β/ERK信号通路有关；在完成项目研究基础上，进一步拓展了主要的活性黄酮化合物药代动力学及其代谢产物研究。