Vitamin E Can Ameliorate Oxidative Damage of Ovine Hepatocytes In Vitro by Regulating Genes Expression Associated with Apoptosis and Pyroptosis, but Not Ferroptosis

• Published in: Molecules (Basel, Switzerland) Vol. 26; no. 15; p. 4520
• Main Authors: Jian, Luyang, Xue, Ying, Gao, Yuefeng, Wang, Bo, Qu, Yanghua, Li, Shuanghong, Li, Heqiong, Li, Zhen, Wang, Bing, Luo, Hailing
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 27.07.2021; MDPI
• Subjects: Antioxidants; Apoptosis; Cell death; Cholecystokinin; Cytokines; Ferroptosis; Gene expression; Genes; Hepatocytes; Hydrogen peroxide; IL-1β; Intracellular; IRAK protein; Kinases; Lipids; Liver; Metabolism; Mortality; non-antioxidation; Oxidative stress; Pretreatment; Proteins; Pyroptosis; Sheep; Survival; Testes; TLR7 protein; Tocopherol; Toll-like receptors; Vitamin E
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules26154520

(1) Background: the current research was conducted to investigate the potential non-antioxidant roles of vitamin E in the protection of hepatocysts from oxidative damage. (2) Methods: primary sheep hepatocytes were cultured and exposed to 200, 400, 600, or 800 μmol/L hydrogen peroxide, while their viability was assessed using a CCK-8 kit. Then, cells were treated with 400 μmol/L hydrogen peroxide following a pretreatment with 50, 100, 200, 400, and 800 μmol/L vitamin E and their intracellular ROS levels were determined by means of the DCF-DA assay. RNA-seq, verified by qRT-PCR, was conducted thereafter: non-treated control (C1); cells treated with 400 μmol/L hydrogen peroxide (C2); and C2 plus a pretreatment with 100 μmol/L vitamin E (T1). (3) Results: the 200–800 μmol/L hydrogen peroxide caused significant cell death, while 50, 100, and 200 μmol/L vitamin E pretreatment significantly improved the survival rate of hepatocytes. ROS content in the cells pretreated with vitamin E was significantly lower than that in the control group and hydrogen-peroxide-treated group, especially in those pretreated with 100 μmol/L vitamin E. The differentially expressed genes (DEGs) concerning cell death involved in apoptosis (RIPK1, TLR7, CASP8, and CASP8AP2), pyroptosis (NLRP3, IL-1β, and IRAK2), and ferroptosis (TFRC and PTGS2). The abundances of IL-1β, IRAK2, NLRP3, CASP8, CASP8AP2, RIPK1, and TLR7 were significantly increased in the C1 group and decreased in T1 group, while TFRC and PTGS2 were increased in T1 group. (4) Conclusions: oxidative stress induced by hydrogen peroxide caused cellular damage and death in sheep hepatocytes. Pretreatment with vitamin E effectively reduced intracellular ROS levels and protected the hepatocytes from cell death by regulating gene expression associated with apoptosis (RIPK1, TLR7, CASP8, and CASP8AP2) and pyroptosis (NLRP3, IL-1β, and IRAK2), but not ferroptosis (TFRC and PTGS2).