Structure-dependent induction of apoptosis by hepatotoxic pyrrolizidine alkaloids in the human hepatoma cell line HepaRG: Single versus repeated exposure.
Structure-dependent induction of apoptosis by hepatotoxic pyrrolizidine alkaloids in the human hepatoma cell line HepaRG: Single versus repeated exposure.
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DOI：
10.1016/j.fct.2018.02.036
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发表时间：
2018-02
期刊：
影响因子：
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通讯作者：
J. Waizenegger;A. Braeuning;M. Templin;A. Lampen;S. Hessel-Pras
中科院分区：
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作者：
J. Waizenegger;A. Braeuning;M. Templin;A. Lampen;S. Hessel-Pras
吡咯里西啶生物碱 (PA) 是次生植物化合物。人类 PA 中毒会导致严重的急性和慢性肝毒性。然而，PA对人类肝毒性的分子机制尚不清楚。因此，我们研究了用结构不同的逆连碱（echimidine、千里光碱）、天芥菜碱（heliotrine）和otonecine型（senkirkine）的PA单次（24小时）或重复剂量（14天）处理后人类HepaRG细胞的细胞死亡参数。暴露24小时后，只有retronecine型PA在HepaRG细胞中具有细胞毒性，并诱导细胞凋亡，表现为膜不对称性的丧失、线粒体膜电位的破坏以及半胱天冬酶原和PARP裂解的增加。相比之下，14天后所有四种PA都发挥了上述效果。此外，仅在单次暴露（6小时）后，retronecine型PA才检测到凋亡事件caspase 3、8和9激活以及核浓缩和DNA断裂。总的来说，我们的研究揭示了PA暴露后时间和结构依赖性的细胞凋亡，表明retronecine型PA似乎是比天芥菜碱或otonecine型PA更有效的细胞凋亡诱导剂。此外，我们的结果表明，PA 诱导的 HepaRG 细胞凋亡最有可能通过涉及外在死亡受体途径和内在线粒体途径发生。
Pyrrolizidine alkaloids (PA) are secondary plant compounds. PA intoxication in humans causes severe acute and chronic hepatotoxicity. However, the molecular mechanisms of PA hepatotoxicity in humans are not well understood yet.Therefore, we investigated cell death parameters in human HepaRG cells following either single (24 h) or repeated dose treatment (14 d) with structurally different PA of the retronecine (echimidine, senecionine), heliotridine (heliotrine), and otonecine type (senkirkine). After 24 h of exposure only retronecine-type PA were cytotoxic in HepaRG cells and induced apoptosis indicated by a loss of membrane asymmetry, disruption of the mitochondrial membrane potential, and increased pro-caspase and PARP cleavage. In contrast, after 14 d all four PA exerted the aforementioned effects. Furthermore, the apoptotic events caspase 3, 8 and 9 activation as well as nuclear condensation and DNA fragmentation were only detected for the retronecine-type PA after single exposure (6 h).Overall, our studies revealed a time- and structure-dependent apoptosis after PA exposure, suggesting that retronecine-type PA seem to be more potent apoptosis inducers than heliotridine- or otonecine-type PA. Furthermore, our results suggest that PA-induced apoptosis in HepaRG cells occur most probably by involving both, the extrinsic death receptor pathway as well as the intrinsic mitochondrial pathway.