基于谱效（毒）相关和PI3K/Akt/mTOR信号通路介导的何首乌肝毒性物质基础及作用机制研究

Polygonum multiflorum (PM) is a commonly used traditional Chinese medicine and has a long use history in China. However, hepatic toxicity of PM has been reported frequently both at home and abroad in recent years, which has led to the clinical value of PM under suspicion. Therefore, it has great significance to elucidate the hepatotoxic substance basis and action mechanism of PM. Recent studies have suggested that PI3K/Akt/mTOR signaling pathway plays an important role in drug-induced liver injury. Many drug components, especially anthraquinones, can modulate this signaling pathway, leading to hepatocyte apoptosis and autophagy, however, it is not clear whether the hepatotoxicity of PM is related to this pathway. Firstly, Based on the previous work and the understanding of the pathogenesis of drug-induced liver injury in modern medicine, this project used UHPLC-Q-TOF/MS qualitative and quantitative analysis methods to perform fingerprint analysis on the chemical composition of different processed products of PM, combining with liver toxicity evaluation indicators, then via spectral-effect (toxicity) correlation analysis to identify the hepatotoxic components of PM. Secondly, the extract PM and its toxic components were applied to the SD rats and normal human LO2 hepatocyte model, by analyzing the expression of PI3K/Akt/mTOR signaling pathway related proteins in vitro and in vivo, to explore the mechanism of PM hepatotoxicity and identify the hepatotoxic substance basis and target of action. This project can standardize the processing technology and provide theoretical and experimental basis for the safety of clinical medication.

何首乌为临床常用中药，在我国使用历史悠久，但近年来国内外频繁报导其具肝毒性，导致何首乌临床价值受到严重质疑，因此，阐明何首乌肝毒性物质基础及作用机制具有重要意义。近年研究提示PI3K/Akt/mTOR信号通路在药物性肝损伤方面起到重要作用，多种药物成分，尤其蒽醌类成分可调节该通路，从而引起肝细胞凋亡与自噬，但何首乌肝毒性的产生是否与该通路有关尚不清楚。故本课题结合现代医学对药物性肝损伤发病机理的认识，采用HPLC-Q-TOF/MS定性定量分析方法对何首乌不同炮制品进行化学成分指纹分析，结合肝毒性评价指标，通过谱效（毒）相关分析，辨识何首乌肝毒性成分。然后将（制）何首乌及毒性成分作用于SD大鼠和正常人LO2肝细胞模型，采取激活和抑制PI3K/Akt/mTOR信号通路相关蛋白的表达，探讨何首乌致肝毒性机制，明确肝毒性物质基础和作用靶标，为规范何首乌炮制工艺，确保临床用药安全，提供科学依据。

何首乌为临床常用中药，在我国使用历史悠久，但近年来国内外频繁报导其具肝毒性，导致何首乌临床价值受到严重质疑。本项目采用HPLC-Q-TOF/MS定性定量分析方法对何首乌不同炮制品进行化学成分指纹分析，结合肝毒性评价指标，辨识何首乌肝毒性成分。以正常人LO2肝细胞模型，采取激活和抑制PI3K/Akt/mTOR信号通路相关蛋白的表达，探讨何首乌致肝毒性机制，明确肝毒性物质基础和作用靶标，为规范何首乌炮制工艺，确保临床用药安全，提供科学依据。目前明确了不同炮制工艺可以显著影响何首乌中主要成分二苯乙烯苷（TSG）的含量，以及TSG对PI3K/Akt/mTOR信号通路介导的肝细胞自噬作用，发现miR-122和miR-194能够被TSG诱导而上调表达，且文献报道miR-122和miR-194可能作为肝毒性的生物标志物，本项目整体已达到预期研究目标。