Identification of the structure-dependent toxicity of hepatotoxic pyrrolizidine alkaloids

肝毒性吡咯里西啶生物碱的结构依赖性毒性的鉴定

• 批准号: 315853093
• 负责人: Professor Dr. Alfonso Lampen
• 金额: --
• 依托单位: Abteilung 5: Lebensmittelsicherheit
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/315853093?language=en
• 关键词: Identification; structure; dependent; toxicity; hepatotoxic

Unsaturated pyrrolizidine alkaloids (PA) belong to the most toxic compounds; more than 400 PA have been described in a wide range of plants. They are also found in Senecio vulgaris L. PA are toxic to humans and animals. PA were detected in substantial amounts into honey and tea. Acute PA poisoning via food contamination causes severe damage to the liver; long-term, sub-lethal doses may cause cumulative damage or cancer. Animal studies revealed that PA are carcinogenic. Mutagenicity of PA was observed in several in vitro and in vivo test systems. The toxicity of these compounds is associated with enzymatic reactions by enzymes dominantly expressed in the liver and dependent on PA´s bioavailability. However, the mode of action of PA to generate hepatotoxicity is not yet identified.Based on our results for bioavailability and the molecular mode of action of the structurally different PA echimidine, heliotrine, senecionine, and senkirkine we hypothesize that PA act in a structure-dependent manner. Therefore we aimed to group the PA structure- and toxicity-dependent within this project.For that purpose bioavailability shall be further studied in an in vitro model for the human intestinal barrier with a larger set of PA (22). As endpoint for grouping the ABCB1-dependent transport/detoxification shall be investigated for the structure-dependent mode of action of PA. Additionally, the molecular mechanism of PA hepatic toxicity shall be analyzed. We already showed in our transcriptomic study with the 4 structurally different PA differences in the induction of gene expression in the liver. Therefore, in vivo analysis of the induction of hepatotoxicity in mice in real time shall be analysed using two-photon excitation microscopy. The structure-dependent induction of toxicity shall be investigated using specific liver marker in wildtype mice and in further step in Abcb1 knockout mice. The identified toxic effects on liver shall be correlated with the transcriptomics data from human primary hepatocytes. Based on these results specific genes and signalling pathways shall be verified in regard to a structure-dependent induction by PA using quantitative real time PCR and reporter gene assays, respectively.This study aimed to close gaps in regard to the structure-dependent PA bioavailability and hepatic mode of action to form PA toxicity group on the basis of its different structure. These data shall contribute to a better understanding of the toxic consequences of an acute or chronic PA uptake to enhance the protection of humans.

不饱和吡咯lizidine生物碱（PA）是毒性最大的一类化合物；在广泛的植物中发现了400多种PA。它们也被发现在Senecio vulgaris L. PA对人类和动物有毒。蜂蜜和茶叶中检测到大量的PA。食物污染引起的急性PA中毒对肝脏造成严重损害；长期的亚致死剂量可能导致累积损害或癌症。动物实验表明PA具有致癌性。在体外和体内试验系统中观察了PA的致突变性。这些化合物的毒性与主要在肝脏中表达的酶的酶促反应有关，并依赖于PA的生物利用度。然而，PA产生肝毒性的作用方式尚未确定。基于我们的生物利用度和结构不同的PA的分子作用模式，我们假设PA以结构依赖的方式起作用。因此，我们的目标是在这个项目中对PA的结构和毒性进行分组。为此，生物利用度需要在具有更大PA组的人肠屏障体外模型中进一步研究（22）。作为分组abcb1依赖性转运/解毒的终点，应研究PA的结构依赖性作用模式。并对PA肝毒性的分子机制进行分析。我们已经在转录组学研究中证明了4种结构不同的PA在肝脏中诱导基因表达的差异。因此，对小鼠肝毒性诱导的实时体内分析应采用双光子激发显微镜进行分析。结构依赖性的毒性诱导将在野生型小鼠和Abcb1敲除小鼠中使用特异性肝脏标志物进行进一步研究。已确定的对肝脏的毒性作用应与人原代肝细胞的转录组学数据相关。基于这些结果，将分别使用实时定量PCR和报告基因检测来验证PA对结构依赖性诱导的特定基因和信号通路。本研究旨在弥补结构依赖性PA生物利用度和肝脏作用方式的空白，根据其不同的结构形成PA毒性基团。这些数据将有助于更好地了解急性或慢性PA摄取的毒性后果，以加强对人类的保护。

项目成果

Structure-dependent induction of apoptosis by hepatotoxic pyrrolizidine alkaloids in the human hepatoma cell line HepaRG: Single versus repeated exposure.

• DOI: 10.1016/j.fct.2018.02.036
• 发表时间: 2018-02
• 期刊: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association
• 作者: J. Waizenegger;A. Braeuning;M. Templin;A. Lampen;S. Hessel-Pras

The pyrrolizidine alkaloid senecionine induces CYP-dependent destruction of sinusoidal endothelial cells and cholestasis in mice

• DOI: 10.1007/s00204-019-02582-8
• 发表时间: 2019-10-12
• 期刊: ARCHIVES OF TOXICOLOGY
• 影响因子: 6.1
• 作者: Hessel-Pras, Stefanie;Braeuning, Albert;Reif, Raymond
• 通讯作者: Reif, Raymond

Comparative analysis of 3D culture methods on human HepG2 cells

• DOI: 10.1007/s00204-016-1677-z
• 发表时间: 2017-01-01
• 期刊: ARCHIVES OF TOXICOLOGY
• 影响因子: 6.1
• 作者: Luckert, Claudia;Schulz, Christina;Hessel, Stefanie
• 通讯作者: Hessel, Stefanie