行气活血法调控枯否细胞中PPAR-γDNA甲基化干预早期肝纤维化的机制研究

In the early stage of hepatic fibrosis(HF), it is an important treatment method to cure qi stagnation with removing stasis medicinal herbs. The fruits of Euscaphisfukienensis Hsu. own the function of warming the middle jiao and regulating qi. In our previous studies, the fruits showed good effect on anti-hepatic fibrosis, and the triterpenoids(EFT) of the fruits prevent the kupffer cell(KC)to polarize to M1 macrophage. We suppose the fruit should show better effect on anti-hepatic fibrosis with Danshen, which is a good medicinal herbs with improving circulation of blood. Activation of hepatic stellate cell (HSC) is the key chain in HF. KC plays a important role in activating HSC. Under induction of LPS, KC is polarized to M1 macrophage to produce pro-imflammatory factors to activate HSC. NF-κB is closely related to KC polarization. NF-κB is the critical downstream effector protein in TLR pathway. In physiological status, TLR pathway is inhibited by PPAR-γ, but in early stage of HF, PPAR-γ gene is in silence expression. Gene expression is controlled by DNA methylation. So we assume there is a relationship between the PPAR-γ DNA methylation and KC polarization. Methylation of PPAR-γ DNA inhibits the express of PPAR-γ in KC , activates the NF-κB and promotes the polarization of KC to M1 macrophage. The fruits of Euscaphisfukienensis Hsu. and Danshen and their effective constituents (EFT and phenolic acids of Danshen)will prevent the methylation of PPAR-γ DNA , inhibit the NF-κB to stop the polarization of KC, and finally prevent the activation of HSC. The results provide the experimental basis for the fruits of Euscaphisfukienensis Hsu. and Danshen and their effective constituents in the treatment of hepatic fibrosis, and also explain the scientific intension of Xinqinhuoxue method in treatment HF.

早期肝纤维化（HF）时以疏肝理气为枢机，配合活血化瘀药是治疗其有效方法。前期研究发现，具有理气止痛的福建野鸦椿果有较好的抗HF效果，其所含三萜能抑制枯否细胞（KC）M1极化，辅以丹参将增加其抗HF效果。KC在LPS诱导下极化成M1型，分泌促炎细胞因子，促使HSC增殖活化。KC表型变化与NF-κB有关，而NF-κB是TLR信号通路下游蛋白，生理状态下TLR 信号可由PPAR-γ负调控。HF发生时PPAR-γ 表达沉默，而基因表达沉默与DNA甲基化有关。因此我们提出福建野鸦椿果与丹参及其有效组分能抑制PPAR-γ DNA甲基化，增加PPAR-γ的表达，抑制NF-κΒ活性，干预KC M1极化， 降低HSC 的增殖与激活。本项目通过体内外实验从PPAR-γ DNA甲基化对KC极化影响探讨行气活血法指导下的药物作用机制，研究结果不仅为临床用药提供参考，也将从新角度阐明行气活血法治疗HF的科学内涵。