The NTCP receptor inhibitor Ezetimibe reduces the toxicity of α-amanitin: in vitro and in vivo study.

Background and Aims: To verify the protective effect of Ezetimibe, an sodium taurocholate co-transporting polypeptide (NTCP) inhibitor, on α-amanitin poisoning in vitro and in vivo by inhibiting NTCP to prevent α-amanitin into hepatocytes. Approach and Results: In animal experiments, the survival rate was significantly improved in the treatment group. The pathomorphological characteristics of liver and kidney in the treatment group were significantly improved. In cell experiments,The cell viability of the treatment group was significantly improved, and the expression of NTCP in the treatment group was significantly decreased by immunofluorescence. In molecular docking simulations, we demonstrated the potential of NTCP to bind Ezetimibe and α-amanitin, respectively. Transcriptomics in high-throughput sequencing was used to detect the differential metabolic genes between α-amanitin poisoning group and the treatment group, and signal pathway enrichment was used to analyze the significantly different signal pathways. Conclusions: Ezetimibe, as an inhibitor of NTCP, can reduce the entry of α-amanitin into hepatocytes to play a protective role and improve the cell viability and survival rate of mice. Compare the control group using high-throughput sequencing Differences in mRNA gene expression in L-02 cell lines between α-amanitin poisoning group and Ezetimibe treatment group after poisoning

研究背景与目的：本研究旨在验证依泽替米贝（Ezetimibe）——一种牛磺胆酸钠协同转运多肽（sodium taurocholate co-transporting polypeptide, NTCP）抑制剂——通过抑制NTCP阻止α-鹅膏毒肽（α-amanitin）进入肝细胞，从而在体内外模型中对α-鹅膏毒肽中毒发挥保护作用。研究方法与结果：动物实验中，治疗组小鼠的生存率显著提升；治疗组肝肾组织的病理形态学特征得到明显改善。细胞实验显示，治疗组细胞活力显著升高，免疫荧光检测结果表明治疗组NTCP的表达量显著下调。分子对接模拟实验证实，NTCP可分别与依泽替米贝、α-鹅膏毒肽结合。本研究通过高通量转录组测序检测α-鹅膏毒肽中毒组与治疗组间的差异代谢基因，并通过信号通路富集分析筛选显著差异的信号通路。研究结论：依泽替米贝作为NTCP抑制剂，可通过减少α-鹅膏毒肽进入肝细胞发挥保护作用，提升细胞活力与小鼠生存率。本研究通过高通量测序，对比L-02细胞系中对照组、α-鹅膏毒肽中毒组与中毒后依泽替米贝治疗组的mRNA基因表达差异。