基于骨髓间充质干细胞抑制肝星状细胞活化研究一贯煎逆转肝纤维化的机制

Liver fibrosis is the common pathologic change of all kinds of chronic liver diseases. liver fibrosis reversion is the key for treating chronic liver diseases . Yin deficiency of liver and kidney is the important pathogenesis of liver fibrosis, The last country natural science foundation research confirmed Yiguanjian has good effect on liver fibrosis reversion by Bone marrow mesenchymal stem cells inducing hepatic stellate cell apoptosis. However, the study found: A few apoptosis of hepatic stellate cells cannot explain the whole mechanism of liver fibrosis reversal. The activated hepatic stellate cells turn into a quiescent state play a more important role, Signaling pathway FGF2－Dlk1－Notch1 is the key way. Based on the observation of the evolution law of yin deficiency syndrome on liver fibrosis, in this study, we used liver fibrosis rat model, on the whole animals in vivo and cells in vitro, The HSCs activation was observed. FGF2 was detected with ELISA. The expression of mRNA and protein of typeⅠ/Ⅲ collagen protein、Dlk1、Notch1 were determined by Real-time PCR and western blotting respectively. We will explore the mechanism of Yiguanjian to reverse the liver fibrosis. This research has important meaning for explaining microcosmic pathologic basis of yin deficiency of liver and kidney and pharmacological mechanism of Yiguanjian.

肝纤维化是多数慢性肝损伤的共同病理，逆转肝纤维化是其治疗的关键。肝肾阴虚是该病的重要病机之一。前期国家自然科学基金研究证实，滋补肝肾中药一贯煎通过骨髓间充质干细胞诱导肝星状细胞凋亡发挥逆转肝纤维化的作用。然研究发现：少数肝星状细胞的凋亡，不能解释一贯煎逆转肝纤维化的全部机制，活化的肝星状细胞转变为静止状态发挥更为重要的作用。FGF2－Dlk1－Notch1信号通路是其关键途径。本课题在观察肝纤维化阴虚证演变规律基础上，建立骨髓移植+CCl4诱导的肝纤维化大鼠模型，结合细胞实验，观察一贯煎干预FGF2－Dlk1－Notch1信号通路的机制：ELISA法检测肝组织FGF2的含量；实时荧光定量PCR、蛋白质印迹技术检测Dlk1、Notch1等mRNA转录水平及蛋白表达，同时观察肝星状细胞活化状态，探讨一贯煎逆转肝纤维化的机制。本课题对阐释肝肾阴虚证病理基础和一贯煎逆转肝纤维化的机制有重要意义。

肝纤维化是指肝组织内细胞外基质成分过度增生与异常沉积，导致肝脏结构和功能异常的病理变化。肝纤维化见于大多数不同病因的慢性肝脏疾病中，如慢性乙型肝炎、酒精性肝炎、脂肪肝等疾病在其病变过程中均会有肝纤维化的存在，如果不积极治疗可进一步发展形成肝硬化，严重影响患者健康和生命。目前，现代医学对该病的治疗主要是针对病因和抗肝纤维化的治疗，但多属药物仍处于实验研究阶段，目前尚无满意的治疗药物。中医学应用滋补肝肾法治疗肝纤维化在临床上获得了显著的疗效。为了进一步发挥中药的作用，揭示其作用机制是十分重要的环节。课题基于肾主骨生髓、肝肾之阴调控肝星状细胞活化状态这一理论基础，制备与人类肝纤维化形态特征相似、恢复期相对较短的CCl4诱导的肝纤维化大鼠模型，结合骨髓间充质干细胞-肝星状细胞共培养方法，从整体实验和离体实验两个方面阐释一贯煎逆转肝纤维化的分子机制。研究结果显示，一贯煎逆转肝纤维化效果良好，不仅能够改善肝纤维化大鼠阴虚证表征，还能够显著减轻肝纤维化的程度。其作用机制是一贯煎通过FGF2－Dlk1－Notch1信号通路促进骨髓干细胞抑制肝星状细胞的活化状态而实现的。课题对阐释肝肾阴虚病理基础和一贯煎药理机制有重要意义。