Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, Liaoning, PR China.
Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, Liaoning, PR China; Department of Chemistry and Life Science, School of Advanced Engineering, Kogakuin University, 2665-1, Nakanomachi, Hachioji, Tokyo 192-0015, Japan; Nippi Research Institute of Biomatrix, Toride, Ibaraki 302-0017, Japan.
Toxicol In Vitro. 2022 Apr;80:105330. doi: 10.1016/j.tiv.2022.105330. Epub 2022 Feb 11.
Silibinin is a natural polyphenolic flavonoid, isolated from the seeds of the milk thistle of Silybum marianum (L.) Gaertn. Silibinin has been widely used clinically as a traditional medicine for liver diseases. This study investigated the protective role of silibinin in ethanol- or acetaldehyde-induced apoptosis in human carcinomatous liver HepG2 cells and immortalized liver HL7702 cells, focusing on elucidation of the underlying mechanism in vitro. The toxicity of ethanol or acetaldehyde was evaluated by MTT assay. Apoptosis-related proteins, mitochondrial fission-associated proteins and mitochondrial fusion-associated proteins were analyzed by western blotting and immunofluorescence microscopy. Present experimental results demonstrated that silibinin improved cell viability, reduced the enzyme activities of AST/ALT and ALDH/ADH, inhibited apoptosis and recovered mitochondrial function in ethanol- or acetaldehyde-treated HepG2 or HL7702 cells. Silibinin reduced the expression of mitochondrial fission-associated proteins, dynamin-related protein 1 (DRP1), but increased mitochondrial fusion-associated proteins, optic atrophy 1 (OPA1) and mitofusin 1 (MFN1). Accordingly, inhibition of DRP1 activity with its pharmacological inhibitor or siDRP1 efficiently attenuated ethanol- or acetaldehyde-induced apoptosis, whereas activation of DRP1 by using staurosporine (STS) further increased apoptosis in ethanol- or acetaldehyde-treated HepG2 or HL7702 cells. The results show that silibinin protects cells against ethanol- or acetaldehyde-induced mitochondrial fission that results in apoptosis.
水飞蓟宾是一种天然多酚类黄酮，从奶蓟草（Silybum marianum（L.）Gaertn）的种子中分离得到。水飞蓟宾已广泛临床用于治疗肝脏疾病的传统药物。本研究探讨了水飞蓟宾在乙醇或乙醛诱导的人肝癌 HepG2 细胞和永生化肝 HL7702 细胞凋亡中的保护作用，重点在于体外阐明其潜在机制。通过 MTT 测定评估乙醇或乙醛的毒性。通过 Western blot 和免疫荧光显微镜分析凋亡相关蛋白、线粒体分裂相关蛋白和线粒体融合相关蛋白。目前的实验结果表明，水飞蓟宾可提高细胞活力，降低 AST/ALT 和 ALDH/ADH 酶活性，抑制乙醇或乙醛处理的 HepG2 或 HL7702 细胞凋亡并恢复线粒体功能。水飞蓟宾降低了线粒体分裂相关蛋白、动力相关蛋白 1（DRP1）的表达，但增加了线粒体融合相关蛋白、视神经萎缩 1（OPA1）和线粒体融合蛋白 1（MFN1）的表达。因此，用其药理学抑制剂或 siDRP1 抑制 DRP1 活性可有效减轻乙醇或乙醛诱导的凋亡，而用 staurosporine（STS）激活 DRP1 则进一步增加乙醇或乙醛处理的 HepG2 或 HL7702 细胞的凋亡。结果表明，水飞蓟宾可防止细胞发生乙醇或乙醛诱导的线粒体分裂导致的凋亡。
