Multilevel molecular analysis of the hepatoprotective effect of medicinal herbs extracts in prevention of liver dysfunction caused by aflatoxin B1 in pig as an animal model (in-vivo), and hepatocyte cell culture analysis in human and pig (invitro)

本课题旨在研究中药对黄曲霉毒素B1（AFB 1）肝毒性的保护作用，并与体外培养的人肝细胞进行比较。为了精确和全面的阐明，在转录组、蛋白质组和代谢组的水平上进行多层次的分子分析。该项目的动机来自于经常发生的饲料和食品黄曲霉毒素，特别是黄曲霉毒素B1的意外污染。我们假设，对肝细胞有有益作用的保肝草药[穿心莲AP，水飞蓟SM，姜黄CL]也可以减轻黄曲霉毒素B1污染的饮食的不良影响。在一个具有亚临床症状的试点项目中指定的肝毒性剂量的黄曲霉毒素B1与草药提取物（WP 1）组合喂养。通过组织学和（实验室）临床研究（WP 1）以及体内蛋白质组（WP 2）、线粒体功能和结构变异（WP 3）、miRNA-（WP 4）和mRNA-（WP 6，WP 7）表达和代谢组（WP 8）分析来表征黄曲霉毒素B1（AFB 1）的肝毒性作用和草药提取物对肝功能的保护作用。人（THLE 2）和猪（JSNK-1）细胞的体外实验通过表达分析和siRNA实验（WP 8）支持了各种分子途径的偏移和关键基因的特定功能作用。通过对所有数据进行全面的生物信息学整合和分析，确保在分子水平上检测详细变化并解释数据（WP 9）。广泛的现代分子方法提供了对黄曲霉毒素反应的生物和代谢过程的根本变化的见解，并结合作为促进健康的膳食补充剂提供的草药的潜在保肝作用。该研究提供了与猪饲料设计相关的结果，以及是否可以预防意外黄曲霉毒素污染的影响。此外，通过比较人类和猪细胞的反应，该研究可以说明在健康饮食的意义上使用草药的潜力。

The project aims to investigate the hepatoprotective effect of medical herbs on hepatotoxic effects of aflatoxin B1 (AFB1) in vivo in piglets and in comparison in human hepatocyte cultures in vitro. For precise and comprehensive elucidation, a multilevel molecular analysis is performed at the levels of the transcriptome, proteome and metabolome. The motivation for the project arose from recurrent accidental contamination of feed and food with aflatoxin, especially AFB1. We hypothesised that hepatoprotective medical herbs [Andrographis paniculata AP, Silybum Marianum SM, Curcuma Longa CL], which have a beneficial effect on liver cells, could also attenuate the adverse effects of a diet contaminated with AFB1. A hepatotoxic dose of AFB1, specified in a pilot project with subclinical symptoms is fed in combination with extracts of the medical herbs (WP1). The hepatotoxic effect of aflatoxin B1 (AFB1) and the hepatoprotective effect of medical herb extracts on liver function are characterized by histological and (laboratory) clinical investigations (WP1) as well as analysis of the proteome (WP2), the mitochondrial functional and structural variation (WP3), the miRNA- (WP4) and mRNA- (WP6, WP7) expression and the metabolome (WP8) in vivo. In vitro experiments with human (THLE2) and porcine (JSNK-1) cells underpin the excursion of various molecular pathways and the specific functional role of key genes by expression analysis and siRNA experiments (WP8). The detection of detailed changes at the molecular level and interpretation of the data is ensured by a comprehensive bioinformatics integration and analysis of all data (WP9). A wide range of modern molecular methods provides insights into the fundamental changes in biological and metabolic processes in response to aflatoxins and in combination with the potential hepatoprotective effects of medical herbs offered as health-promoting dietary supplements. The study provides results relevant to the design of diets for pigs and whether the effects of accidental aflatoxin contamination can be prevented. Furthermore, by comparing the reaction of human and porcine cells, the study allows statements to be made about the potential of using medical herbs in the sense of a healthy diet.