Özerkan Dilşad, Özsoy Nesrin, Yılmaz Erkan
Department of Biology, Faculty of Arts and Sciences, Kastamonu University, 37100, Kastamonu, Turkey.
Department of Biology, Faculty of Science, Ankara University, Tandogan, 06100, Ankara, Turkey.
Cytotechnology. 2015 Dec;67(6):995-1002. doi: 10.1007/s10616-014-9738-8. Epub 2014 Jul 6.
Carbon tetrachloride (CCl4) is widely used to induce liver toxicity in in vitro/in vivo models. Lipid peroxidation (LPO) begins with toxicity and affects cell viability. Recently, the beneficial effects of melatonin and Vitamin D on cell proliferation in human normal and cancer cells were found. This study was planned to evaluate antioxidant and cytoprotective activity of melatonin and Vitamin D in CCl4 induced cytotoxicity in HepG2 and Hep3B hepatoma cell lines. Based on the cytotoxicity assay, melatonin and Vitamin D were evaluated for cytotoprotective potential against CCl4 induced toxicity in HepG2 and Hep3B liver cell lines by monitoring cell viability, LPO and glutathione (GSH) level. Different dosages of CCl4 (0.1, 0.2, 0.3 and 0.4 % v/v) were applied to HepG2 and Hep3B cells in order to determine the most toxic dosage of it in a time dependent manner. The same experiments were repeated with exogenously applied melatonin (MEL) and Vitamin D to groups treated with/without CCL4. Cell viability was determined with MTT measurements at the 2nd, 24th and 48th h. GSH content and Malondialdehyde levels were measured from the cell lysates. As a result, both melatonin and Vitamin D administration during CCl4 exposure protected liver cells from CCl4 induced cell damage. Increase in LPO and decrease in GSH were found in the CCl4 groups of both cells. Contrary to these results administration of MEL and Vitamin D on cells exhibited results similar to the control groups. Therefore, melatonin and Vitamin D might be a promising therapeutic agent in several toxic hepatic diseases.
四氯化碳(CCl4)被广泛用于在体外/体内模型中诱导肝毒性。脂质过氧化(LPO)始于毒性作用,并影响细胞活力。最近,发现褪黑素和维生素D对人正常细胞和癌细胞的细胞增殖具有有益作用。本研究旨在评估褪黑素和维生素D在CCl4诱导的HepG2和Hep3B肝癌细胞系细胞毒性中的抗氧化和细胞保护活性。基于细胞毒性试验,通过监测细胞活力、LPO和谷胱甘肽(GSH)水平,评估褪黑素和维生素D对CCl4诱导的HepG2和Hep3B肝细胞系毒性的细胞保护潜力。将不同剂量的CCl4(0.1、0.2、0.3和0.4% v/v)以时间依赖性方式应用于HepG2和Hep3B细胞,以确定其最具毒性的剂量。对用/不用CCL4处理的组外源性应用褪黑素(MEL)和维生素D重复相同的实验。在第2、24和48小时用MTT测量法测定细胞活力。从细胞裂解物中测量GSH含量和丙二醛水平。结果,在CCl4暴露期间给予褪黑素和维生素D均可保护肝细胞免受CCl4诱导的细胞损伤。在两种细胞的CCl4组中均发现LPO增加和GSH减少。与这些结果相反,对细胞应用MEL和维生素D显示出与对照组相似的结果。因此,褪黑素和维生素D可能是几种中毒性肝病中有前景的治疗剂。