基于已知吸收成分调控病理血管生成-肝纤维化并与母方桃红四物汤疗效的比较

It is evident that pathologic angiogenesis in hepatic fibrosis process, which hindered the inversion and recovery of hepatic fibrosis. We discovered that Honghua's absorbed bioactive compound(ABC) HSYA can regulate the tumor angiogenesis effectively. And many researches show that Taohongsiwu Decoction contained Honghua to treat hepatic fibrosis by compatibility in clinic can obtain theraputic effect, but the mechanism is not clear-cut. So we take the activated HSCs and hepatic fibrosis model rats, apply the multiple techniques of RT-PCR, ELISA, Western blot and so on, to study the effects and mechanisms of Taohongsiwu decoction's cleared absorbed bioactive compound(ABC) HSYA and the parent formula on VEGF and its recepters, endothelial cell survived correlation factors, and the pathological angiogenesis and angiogenesis factors and the main VEGF-mediated tyrosine kinase signal pathway on model rats, and demonstrate the hypothesis that drugs can regulate the pathological angiogenesis and related signal pathway to slow down the hepatic fibrosis. And clarify emphasizedly when the dosage of the absorbed compound HSYA is the same as it in formula the effect difference between the absorbed compound and the parent formula, identify that the potential targets and material basis of Taohongsiwu decoction interfered in hepatic fibrosis from the new point of view, and then hope to establish a new way to apply the comparison between ABC and parent formula to clarify the formula's molecule mechanism of multitarget points.

肝纤维化进程中存在明显病理性血管生成，阻碍了肝纤维化的逆转恢复。前期发现活血药红花的吸收生物活性成分(ABC)HSYA可有效调控病理血管生成；而大量研究表明含有红花的桃红四物汤临床上配伍治疗肝纤维化确有疗效，但其确切的作用机制不完全明确。本项目拟利用激活肝星状细胞(HSC)模型和肝纤维化大鼠模型，运用RT-PCR、ELISA、蛋白印迹等方法，比较桃红四物汤与已知吸收成分HSYA对活化HSC分泌血管内皮生长因子(VEGF)及其受体等的作用；对肝纤维化模型大鼠肝组织病理性血管生成、促血管生成因子以及VEGF介导血管内皮存活主要酪氨酸激酶信号途径的影响，论证"该方调控病理血管生成及相关信号通路以减缓肝纤维化进程"的假说；重点阐明HSYA等ABC(剂量等于母方含量)与母方疗效的异同，从新的角度明确其干预肝纤维化的潜在靶标与物质基础，并开辟一条吸收成分与母方疗效相比较阐明方剂多靶点分子机理的新途径。

慢性肝病伴肝纤维化是肝病进展过程的枢纽环节；防治慢性肝病合并肝纤维化并改善肝功能研究是摆在我们面前的重大课题；目前西医对慢性肝病伴肝纤维化的治疗效果不理想，仍缺乏高效、无明显不良反应的西药。因而天然药物特别是中草药及其复方近年来成为临床防治慢性肝病伴肝纤维化研究的热点。寻求切实有效缓解和逆转此病的中药复方与吸收生物活性成分，并阐明作用靶点及作用途径十分迫切。课题选用传统的四氯化碳(CCl4)致小鼠慢性肝病伴坏死性炎症及纤维化动物模型，运用含主要吸收成分HSYA的红花和桃仁提取物组成的桃红颗粒(THKL)及母方桃红四物汤(THSWT)干预，分别检测了动物血清肝功能、肝纤维化相关因子、组织病理、病理血管生成相关基因及蛋白表达、相关信号通路基因及蛋白表达，发现肝脏病理性血管生成与VEGF、KDR、Flt-1等血管生成因子mRNA与蛋白的大量表达以及PI3K-Akt信号通路的激活有关；THSWT 及THKL对CCl4致的慢性肝病伴坏死性炎症及纤维化小鼠有较好的治疗作用，能够改善肝功能、炎症、降低肝纤维化因子的产生，以及阻止病理血管生成相关促进因子VEGF及其受体KDR、Flt-1的释放和表达，抑制PI3K-Akt信号通路激活，从新的病理血管生成角度证实临床广泛而恰当地运用活血中药复方治疗慢性肝病有一定的科学依据。