清肝活血方通过LXR-LPCAT3信号通路抑制内质网应激改善酒精性肝病的机制研究

Alcoholic liver disease (ALD) will become one of major cause of chronic liver diseases in the future. Based on “damp heat and blood stasis” as the core pathogenesis of ALD, the herb formula “Qinggan Huoxue Recipe(QGHXR)” developed by our study group has been proved effective in improving ALD in clinical or animal experiments. Recent studies have shown that liver X receptor (LXR) inhibit endoplasmic reticulum stress through activating lysophosphatidylcholine acyltransferase (LPCAT3), but it is unclear about its role in ALD. Our preliminary experiments indicated that the expression of LXR, LPCAT3 mRNA was obviously decreased in ALD, and QGHXR could increase its expression. Therefore, we put forward the hypothesis: LXR-LPCAT3 signal pathway is involved in the progress of ALD, and QGHXR improves ALD through suppressing ERS by activating LXR-LPCAT3. In order to verify the hypothesis, we would apply the experimental techniques such as CRISPR/CAS9, molecular biology, targeted metabolomics through clinical, animal model and cell models to explore the role of LXR-LPCAT3 signaling pathway in ALD in order to providing new targets and new strategies for the treatment of ALD, as well as the QGHXR mechanism on treating ALD with the aim to elucidate the effect mechanism of Chinese herbal compound in the treatment of ALD and provide scientific basis for its widespread clinical application.

酒精性肝病（ALD）将成为未来肝脏疾病的主要病因之一。课题组基于其“湿热瘀血”病机研制的中药复方清肝活血方在临床和动物试验中均有效改善ALD。新近研究表明肝脏X受体（LXR）激活磷脂酰胆碱酰基转移酶3（LPCAT3）抑制内质网应激（ERS），但在ALD中的作用并不清楚。前期研究发现LXR、LPCAT3mRNA在ALD模型大鼠肝脏表达有所下调，而清肝活血方干预使其上调。因此我们提出LXR-LPCAT3信号通路调控ALD的发病进程，清肝活血方通过激活LXR-LPCAT3抑制ERS从而改善ALD这一假说。基于此，本项目拟通过临床、动物和细胞试验，应用CRISPR/CAS9、分子生物学、靶向代谢组学等实验技术，探讨LXR-LPCAT3信号通路在ALD中的作用，为ALD的治疗提供新靶点和新策略，以及清肝活血方治疗ALD的作用机制，以阐明中药复方治疗ALD的药效机理，为其在临床应用和推广提供科学依据。

酒精性肝病（Alcoholic liver disease，ALD）作为非感染性肝病的代表将取代病毒性肝炎成为未来肝脏疾病的主要病因之一。研究发现肝脏X受体（liver X receptor，LXR）-溶血磷脂酰胆碱酰基转移酶 3（lysophosphatidylcholine acyltransferase3，LPCAT3）通路对促进多不饱和磷脂类物质的形成，抑制内质网应激（endoplasmic reticulum stress，ERS），阻止ALD进展具有重要意义。课题组基于前期研究已证实清肝活血方（Qinggan Huoxue Recipe，QGHXR）能显著改善ALD患者的临床症状。同时动物体内实验也证实QGHXR能促进酒精的毒性代谢物乙醛在体内的蓄积，对肝脏有保护作用。但药物作用机制仍不明确。本课题旨在通过动物和细胞实验研究主要探讨LXR-LPCAT3信号通路在ALD中的关键作用，以及QGHXR对LXR-LPCAT3信号通路相关分子的影响，为阐明QGHXR治疗ALD的机制提供实验依据。主要结果如下：（1）体内实验证实QGHXR及其拆方均可不同程度改善酒精导致的小鼠肝损伤；（2）体外实验证实QGHXR及其拆方可缓解乙醇对肝细胞的损伤；（3）通过动物实验和细胞实验初步探讨QGHXR及其拆方对LXRα-LPCAT3-ERS相关通路的调节作用，实验表明QGHXR能够调控该通路中LXRα和LPCAT3的表达水平，从而改善ERS，减轻肝脏的脂肪变性和炎症，进而改善酒精引起的肝脏损伤，达到保护肝脏的作用。本课题的顺利实施将为深入认识ALD的分子机制、发现新的治疗靶标做出有益尝试，同时对促进中医药防治ALD有重要的基础和应用价值。