25-Hydroxycholesterol 3-Sulfate Recovers Acetaminophen Induced Acute Liver Injury via Stabilizing Mitochondria in Mouse Models

Abstract

Acetaminophen (APAP) overdose is one of the most frequent causes of acute liver failure (ALF). N-acetylcysteine (NAC) is currently being used as part of the standard care in the clinic but its usage has been limited in severe cases, in which liver transplantation becomes the only treatment option. Therefore, there still is a need for a specific and effective therapy for APAP induced ALF. In the current study, we have demonstrated that treatment with 25-Hydroxycholesterol 3-Sulfate (25HC3S) not only significantly reduced mortality but also decreased the plasma levels of liver injury markers, including LDH, AST, and ALT, in APAP overdosed mouse models. 25HC3S also decreased the expression of those genes involved in cell apoptosis, stabilized mitochondrial polarization, and significantly decreased the levels of oxidants, malondialdehyde (MDA), and reactive oxygen species (ROS). Whole genome bisulfite sequencing analysis showed that 25HC3S increased demethylation of 5mCpG in key promoter regions and thereby increased the expression of those genes involved in MAPK-ERK and PI3K-Akt signaling pathways. We concluded that 25HC3S may alleviate APAP induced liver injury via up-regulating the master signaling pathways and maintaining mitochondrial membrane polarization. The results suggest that 25HC3S treatment facilitates the recovery and significantly decreases the mortality of APAP induced acute liver injury and has a synergistic effect with NAC in propylene glycol (PG) for the injury.

High-Dose Acetaminophen as a Treatment for Cancer

“…On the other hand, NAC (100 mg/kg IP) provided no significant protection. The lack of observed hepato-protection of NAC in these pre-clinical models is surprising, yet has similarly been observed in other studies [13,27]. Higher doses of NAC (500-1200 mg/kg) have demonstrated improved efficacy in preventing AAP toxicity in mouse models [28,29], but are well beyond the maximal safe dose in humans [30].…”

“…Numerous other antidotes to AAP toxicity have been evaluated pre-clinically, including 25-hydroxycholesterol-3-sulfate [13] and heparan sulfates [14]. However, these drugs have not been evaluated in human patients for protection against AAP toxicity; fomepizole and NAC are the drugs with the most clinical data supporting their use.…”

“…As discussed above, the data supporting NAC as the standard rescue agent for AAP overdose are based largely on single-arm non-randomized trials [8]. Furthermore, there is a paucity of data comparing the efficacy of NAC to alternative rescue agents that have demonstrated pre-clinical promise, such as heparan sulfates [14], 25-hydroxycholesterol-3sulfate [13], and the CYP2E1 inhibitor fomepizole [26]. Our lab directly compared the most established antidotes to AAP toxicity, NAC and fomepizole, in pre-clinical mouse models.…”

High-Dose Acetaminophen with Concurrent CYP2E1 Inhibition Has Profound Anticancer Activity without Liver Toxicity

“…The relative lack of efficacy of NAC for preventing hepatotoxicity in our models is unexpected given the established role of NAC as an antidote to AAP toxicity (Akakpo et al, 2022;Heard, 2008). Prior pre-clinical studies of NAC yielded mixed results in preventing AAP liver toxicity (Khayyat et al, 2016;Saito et al, 2010;Wang et al, 2021). Khayyat et al showed that NAC (106 mg/kg IP) administered 1.5 hours after AAP (400 mg/kg IP) decreased ALT from 940 (no NAC) to 860 U/L (+ NAC) relative to baseline ALT of 10 U/L (Khayyat et al, 2016).…”

High-Dose Acetaminophen with N-acetylcysteine Rescue Inhibits M2 Polarization of Tumor-Associated Macrophages

“…In vivo, the treatment with high-dose AAP (500 mg/kg) given simultaneously with NAC/PG was well tolerated by mice. Of note, PG, in addition to being a diluent to enhance AAP solubility for in vivo administration, also helps reverse AAP toxicity via CYP2E1 inhibition [15] and thus was included in the rescue cocktail. AAP treatment alone led to prototypical zone 3 necrosis (around the central veins), where CYP2E1 enzymes are most concentrated [16].…”