基于miRNAs 差异研究何首乌效毒转化规律及作用机制

Polygonum multiflorum (PM) is reported a lot as a typical traditional chinese medicines induced liver injury. At present, the material basis and action mechanism of its induced liver toxicity is unclear and lacking of sensitive index to detect. Circulating miRNAs play important roles in the process of drugs induced liver injury which have specificity and sensitivity. The project will explore the intrinsic biological mechanism of PM induced liver injury from miRNAs. The pathological development related PM induced liver injury will be monitored using liver specific miRNA-122 in serum to investigate their relationship between the development process of liver injury and Polygonum multiflorum, to evaluate the.drug safty of PM and the feasibility of specific miRNA-122 as early warning biomarker of PM induced liver injury; The medical process of main components from PM will be monitored to get the main toxic ingredients by liver specific miRNA-122; The miRNAs experssion differences will be analyzed using mice miRNAs chip, and candidate miRNAs and target signal pathway related with PM induced liver injury will be investigated through biological information softwares. Clinical efficacy to toxic transformation law and its potential mechanism of PM induced liver injury can clarify through the whole research, and get early warning biomarker of PM liver injury. It will provide a feasible scheme for evaluation PM.related products preclinical hepatic toxicity, which has potential application prospect.

何首乌是迄今为止报道较多的引发肝损伤的中药之一，目前其致肝损伤的物质基础和作用机制尚不清楚，原因可能是缺乏检测其肝毒性的敏感指标。循环miRNAs参与许多药物肝损伤过程，具有肝特异性和敏感性。本项目从miRNAs角度出发探讨何首乌效毒转化规律及内在生物学机制。利用血清中肝特异miRNA-122监测何首乌引起肝损伤相关的病理发展，评价何首乌用药安全及miRNA-122作为何首乌肝损伤早期预警指标的可行性；研究miRNA-122与何首乌成分用药的关系，筛选何首乌致肝损伤的毒效成分；通过miRNA 芯片对何首乌致肝损伤模型中小鼠血清miRNA表达谱进行分析和筛选，用生物信息学方法分析何首乌致肝损伤相关的靶点信号通路。通过研究阐明何首乌效毒规律和作用机制，得到何首乌肝毒性的早期预警指标，为何首乌相关产品临床前肝毒性评价提供可行性方案，具有潜在的应用前景。

何首乌是迄今为止报道较多的引发肝损伤的中药之一，通过研究发现循环miRNAs参与许多药物肝损伤过程，具有肝特异性和敏感性。利用血浆中肝特异miRNA-122监测何首乌引起肝损伤相关的病理发展，评价何首乌用药安全及miRNA-122可作为何首乌肝损伤早期预警指标的可能性。研究结果发现miRNA-122对普通的何首乌醇提取物灌胃模型有一定敏感性，12周内表现出明显降低趋势，之后逐渐升高并超过对照组；通过miRNA 芯片对何首乌致肝损伤模型筛选和验证出miR-148a-3p、miR-192及miR-3110-3p对不同喂药周期和喂药剂量呈现出不同程度的敏感；另外，通过不同剂量何首乌醇提取物灌胃小鼠6个月动态监测发现，何首乌对小鼠肝的损伤作用呈现一定的时间和剂量依赖关系；当灌胃量达到80g/kg时，肝组织切片分析结果显示肝细胞炎性浸润、脂肪空泡等明显变化。综上所述，低中剂量仅在长期服用（6个月）何首乌醇提物才能引发小鼠模型肝损伤，而大剂量会出现短时间会引发较明显小鼠肝损伤，同时明确通过单一的miRNA-122指标监测何首乌引发普通模型早期预警欠妥，应进一步验证和开发多个miRNA指标共同监测何首乌引发肝损伤。