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

Abstract

(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).

SBFI26 induces triple‐negative breast cancer cells ferroptosis via lipid peroxidation

“…The cells were seeded in 90‐mm dishes, cultured overnight and subsequently treated with 100 μM SBFI26 for 24 h. The control group did not undergo SBFI26 treatment; each consisted of three replicates. After a culture period of 24 h, the cells were collected and preserved by adding a 1 mL TRIzol reagent prior to RNA extraction and sequencing 48,49 …”

“…After a culture period of 24 h, the cells were collected and preserved by adding a 1 mL TRIzol reagent prior to RNA extraction and sequencing. 48,49 The manufacturer's protocol used the TRIzol reagent to extract the total RNA. The NanoDrop 2000 spectrophotometer (Thermo Scientific, USA) was employed to evaluate RNA purity and quantification.…”

Exploring micronutrient supplements in disease conditions: are they effective?

“…Therefore, micronutrients can be used in the treatment of cardiovascular disease, and their supplementation also improves patients with cardiovascular disease and cancers [8,72,73,[100][101][102]. Micronutrient supplementation can also reduce the risk of pre-eclampsia and high blood pressure in pregnant women [103,104] and improve arterial blood pressure in diabetic and non-diabetic patients [105][106][107][108].…”

“…In the area of mental health, micronutrients improve motor and cognitive function in elderly patients with cognitive-related diseases [76], alleviate oxidative damage and reduce stress and anxiety in diabetic patients [31,67,115]. This thus contributes to the recovery from traumatic brain injury [71] and reverses depressive symptoms [8,73,77,101,[116][117][118][119][120].…”

Crosstalk between regulated necrosis and micronutrition, bridged by reactive oxygen species

“…Kang et al showed that lipid peroxidation facilitated caspase-11-dependent GSDMD cleavage and pyroptosis, while administration of vitamin E attenuated lipid peroxidation and resultant pyroptosis in myeloid lineage cells (215). Pretreatment of vitamin E also effectively reduced hydrogen peroxide-induced ROS and protected hepatocytes against pyroptosis (216). For necroptosis, vitamin E destabilized necrosome formation by attenuating ROS, which inhibited SMAC mimetic/TNFαinduced necroptosis in cancer cells (217).…”