Venlafaxine-Induced Cytotoxicity Towards Isolated Rat Hepatocytes Involves Oxidative Stress and Mitochondrial/Lysosomal Dysfunction
Venlafaxine-Induced Cytotoxicity Towards Isolated Rat Hepatocytes Involves Oxidative Stress and Mitochondrial/Lysosomal Dysfunction
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DOI：
10.15171/apb.2016.066
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发表时间：
2016-12-01
影响因子：
3.6
通讯作者：
Eghbal, Mohammad Ali
中科院分区：
文献类型：
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作者：
Ahmadian, Elham;Babaei, Hossein;Eghbal, Mohammad Ali
目的：抑郁症是世界范围内的一种公共疾病。尽管文拉法辛被广泛用于治疗抑郁症，但它与毒性反应的发生有关。因此，本研究的目的是在新鲜分离的大鼠肝细胞模型上探讨文拉法辛诱导肝细胞死亡的机制。分析细胞损伤、活性氧(ROS)生成、脂质过氧化、线粒体膜电位下降、溶酶体损伤、谷胱甘肽(GSH)水平。结果：文拉法辛以浓度依赖的方式诱导ROS形成，细胞内GSH含量减少，GSSG水平升高，溶酶体膜完整性丧失，线粒体膜崩解，最终细胞死亡。N-乙酰半胱氨酸、牛磺酸和Quercetine显著减少文拉法辛诱导的上述细胞事件。此外，自由基清除剂(丁基羟甲苯和α-生育酚)、细胞色素P42E1抑制剂(4-甲基吡唑)、溶酶体促进剂(甲胺和氯喹)、三磷酸腺苷产生剂(L-谷氨酰胺和果糖)和线粒体封孔剂(三氟拉嗪和L-肉碱)显著减少文拉法辛治疗后的细胞毒性、ROS生成和溶酶体渗漏。线粒体功能障碍伴随着线粒体呼吸系统电子传递复合体II和IV的阻断。结论：我们的数据表明文拉法辛诱导肝细胞氧化应激，我们的研究结果为提出线粒体和溶酶体是文拉法辛介导的细胞损伤的主要靶点提供了证据。
Purpose: Depression is a public disorder worldwide. Despite the widespread use of venlafaxine in the treatment of depression, it has been associated with the incidence of toxicities. Hence, the goal of the current investigation was to evaluate the mechanisms of venlafaxine-induced cell death in the model of the freshly isolated rat hepatocytes.Methods: Collagenase-perfused rat hepatocytes were treated with venlafaxine and other agents. Cell damage, reactive oxygen species (ROS) formation, lipid peroxidation, mitochondrial membrane potential decline, lysosomal damage, glutathione (GSH) level were analyzed. Moreover, rat liver mitochondria were isolated through differential centrifugation to assess respiratory chain functionality.Results: Our results demonstrated that venlafaxine could induce ROS formation followed by lipid peroxidation, cellular GSH content depletion, elevated GSSG level, loss of lysosmal membrane integrity, MMP collapse and finally cell death in a concentrationdependent manner. N-acetyl cysteine, taurine and quercetine significantly decreased the aforementioned venlafaxine-induced cellular events. Also, radical scavenger (butylatedhydroxytoluene and a-tocopherol), CYP2E1 inhibitor (4-methylpyrazole), lysosomotropic agents (methylamine and chloroquine), ATP generators (L-gluthamine and fructose) and mitochondrial pore sealing agents (trifluoperazine and L-carnitine) considerably reduced cytotoxicity, ROS generation and lysosomal leakage following venlafaxine treatment. Mitochondrion dysfunction was concomitant with the blockade of the electron transfer complexes II and IV of the mitochondrial respiratory system.Conclusion: Therefore, our data indicate that venlafaxine induces oxidative stress towards hepatocytes and our findings provide evidence to propose that mitochondria and lysosomes are of the primary targets in venlafaxine-mediated cell damage.