Silibinin (Sil) is used as hepatoprotective drug and is approved for therapeutic use in amanitin poisoning. In our study we compared Sil-bis-succinate (Sil_BS), a water-soluble drug approved for i.v.-administration, with Sil solved in ethanol (Sil_EtOH), which is normally used in research.

We challenged monocultures or 3D-microtissues consisting of HepG2 cells or primary hepatocytes with α-amanitin and treated with SIL_BS, SIL_EtOH, penicillin and combinations thereof. Cell viability and the integrity of the microtissues was monitored. Finally, the expression of the transporters OATP1B1 and B3 was analyzed by qRT-PCR.

We demonstrated that primary hepatocytes were more sensitive to α-amanitin compared to HepG2. Primary hepatocytes cultures were protected by Sil_BS and Sil_EtOH independent of penicillin from the cytotoxic effects of α-amanitin. Subsequent studies of the expression profile of the transporters OATP1B1/B3 revealed that primary hepatocytes do express both whereas in HepG2 cells they were hardly detectable.

Our study showed that Sil_BS has significant advantage over Sil_EtOH with no additional benefit of penicillin. Moreover, HepG2 cells may not represent an appropriate model to investigate Amanita phalloides poisoning in vitro with focus on OATP transporters since these cells are lacking sensitivity towards α-amanitin probably due to missing cytotoxicity-associated transporters suggesting that primary hepatocytes should be preferred in this context.

水飞蓟宾（Sil）用作保肝药物，并被批准用于金黄色素中毒的治疗用途。在我们的研究中，我们比较了批准用于静脉给药的水溶性琥珀酸双琥珀酸酯（Sil _BS）和溶解在乙醇中的Sil（Sil _EtOH），后者通常用于研究。

我们用α-amanitin挑战了由HepG2细胞或原代肝细胞组成的单一培养物或3D显微组织，并用SIL _BS，SIL _EtOH，青霉素及其组合处理。监测细胞活力和微组织的完整性。最后，通过qRT-PCR分析转运蛋白OATP1B1和B3的表达。

我们证明，与HepG2相比，原代肝细胞对α-amanitin更为敏感。独立于青霉素的Sil _BS和Sil _EtOH保护原代肝细胞培养物免受α-amanitin的细胞毒性作用。随后对转运蛋白OATP1B1 / B3的表达谱的研究表明，原代肝细胞确实表达了这两种表达，而在HepG2细胞中几乎无法检测到。

我们的研究表明，Sil _BS具有比Sil _EtOH显着的优势，而没有青霉素的其他优势。此外，HepG2细胞可能不是代表以OATP转运蛋白为核心的体外研究伞形毒蝇中毒的合适模型，因为这些细胞对α-amanitin缺乏敏感性，可能是由于缺少与细胞毒性相关的转运蛋白，这表明在这种情况下应首选原代肝细胞。