Basic Medical School, Zunyi Medical University, Zunyi, Guizhou, China.
School of pharmacy, Zunyi Medical University, Zunyi, Guizhou, China.
Toxicol Lett. 2020 Oct 15;333:49-61. doi: 10.1016/j.toxlet.2020.07.024. Epub 2020 Jul 26.
Hepatotoxicity induced by Mylabris has been reported in both clinical and animal experiments. Cantharidin (CTD), the main active compound of Mylabris was responsible for the hepatotoxicity, which aroused widespread concern. However, the mechanism of CTD hepatotoxicity remained unclear. In this study, LO2 cells were exposed to two doses of CTD (6.25 and 25 μM) for 12 h, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were measured. The metabolites in LO2 cells were profiled by LC-MS. Partial least squares discriminant analysis and orthogonal partial least squares discriminant analysis were used for screening potential biomarkers. The MetPA software was used for clustering and pathway analysis. Network pharmacology was used to predict the genes acted with potential biomarkers. Compared with the control group, the levels of ALT, AST, and LDH was significantly increased after CTD treatment. A total of 46 potential biomarkers for hepatotoxicity induced by CTD were identified. And downregulated potential biomarkers reflected the inhibitory effects of CTD toxicity on metabolism of LO2. Moreover, CTD-induced liver toxicity of LO2 cells is mainly related to three pathways: cysteine and methionine metabolism; glutathione metabolism; and glycine, serine, and threonine metabolism. Furtherly, the mRNA expression of CES2, DNMT1, NOS1, NOS3, S1PR2, and CES1 screened by network pharmacology were regulated by CTD. These studies provide valuable mechanistic insights into CTD-associated hepatotoxicity that will aid in the development of therapeutic prevention and treatment options for this liver disease.
斑蝥素所致肝毒性已在临床和动物实验中得到报道。斑蝥素（CTD）是斑蝥的主要活性成分，其导致肝毒性，引起了广泛关注。然而，CTD 肝毒性的机制仍不清楚。在这项研究中，将 LO2 细胞分别暴露于 2 种浓度的 CTD（6.25 和 25μM）12 h，检测细胞上清液中丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）和乳酸脱氢酶（LDH）的水平。采用 LC-MS 对 LO2 细胞中的代谢物进行分析。采用偏最小二乘判别分析（PLS-DA）和正交偏最小二乘判别分析（OPLS-DA）筛选潜在的生物标志物。利用 MetPA 软件进行聚类和通路分析。采用网络药理学预测与潜在生物标志物相关的基因。与对照组相比，CTD 处理后 LO2 细胞中 ALT、AST 和 LDH 的水平显著升高。共鉴定出 46 个与 CTD 诱导的肝毒性相关的潜在生物标志物。下调的潜在生物标志物反映了 CTD 毒性对 LO2 代谢的抑制作用。此外，CTD 诱导的 LO2 细胞肝毒性主要与三条通路有关：半胱氨酸和蛋氨酸代谢；谷胱甘肽代谢；甘氨酸、丝氨酸和苏氨酸代谢。进一步，网络药理学筛选出的 CES2、DNMT1、NOS1、NOS3、S1PR2 和 CES1 等基因的 mRNA 表达受 CTD 调节。这些研究为 CTD 相关肝毒性提供了有价值的机制见解，有助于开发针对这种肝病的治疗预防和治疗选择。
