Solanum tuberosum peel extract attenuates acetaminophen-induced hepatotoxicity through modulation of inflammatory cytokines, apoptotic pathways, and CYP3A4 expression: evidence from molecular docking, Hep G2 cell assays, and an in ovo embryonic model

Abstract

CYP3A4 is the primary cytochrome P450 that plays a secondary role along with CYP2E1 in the oxidation of acetaminophen (APAP) to the toxic metabolite N-acetyl-p-benzoquinone imine (NAPQI). APAP overdosing activates c-Jun N-terminal kinase (JNK), which activates the mitochondrial membrane permeability transition (MPT). This cascade results in disruption of the hepatic homeostasis. Initially, 70 compounds were short-listed and considered for molecular docking with CYP3A4. Solanine, having shown the highest docking score (− 11.2), was considered as a potential drug, and the ethanolic extract of the fruits of Solanum tuberosum was prepared from pulp and peel. Cytotoxicity of the pulp and peel extracts on Hep-G2 cells was assessed by MTT assay, colony formation assay, and DAPI staining. In the next phase, 14-day-old pathogen-free, fertilised chicken eggs were procured and inoculated with 100μL of APAP (0.01 mg/ml concentration), 10% ethanolic extract of Solanum tuberosum (STE), and 70% alcohol (vehicle control) in five challenge sets and three control sets. After 48 h, allantoic fluids were collected for RT-PCR analysis, and histopathological examination was performed for liver, heart, and brain by H&E staining. The peel extract demonstrated a superior safety profile, showing no inhibitory effect on Hep G2 cell proliferation up to 96 h. in ovo administration of STE significantly down-regulated IL-6, IL-10, IL-1β, TGF-β1, CYP3A4, c-Jun-K, and Bax gene expression compared with the APAP control group. Conversely, STE peels up-regulated the anti-apoptotic Bcl-2 gene. Histopathological examination revealed the protective effect of STE peel on hepatocytes against APAP-induced toxicity. Solanum tuberosum peel extract confers significant protection through modulation of pro-inflammatory cytokines, apoptotic markers, and CYP3A4 expression. Solanine–Glutathione synergy demonstrated strong binding affinity with CYP3A4, enhancing detoxification against APAP-induced hepatotoxicity. Selective cyto-protection by STEpeel extract preserved Hep G2 viability, reduced nuclear damage, and showed superior safety compared to pulp extract. Significant down-regulation of pro-inflammatory cytokines (IL-1β, IL-6, TGF-β1) established potent immunomodulatory effects. STE extracts suppressed pro-apoptotic (Bax, Caspase-3, JNK) and upregulated anti-apoptotic (Bcl-2) genes, protecting hepatocytes. Histopathology confirmed STE, especially peel extract, mitigated APAP-induced necrosis in liver, brain, and heart. Solanine–Glutathione synergy demonstrated strong binding affinity with CYP3A4, enhancing detoxification against APAP-induced hepatotoxicity. Selective cyto-protection by STEpeel extract preserved Hep G2 viability, reduced nuclear damage, and showed superior safety compared to pulp extract. Significant down-regulation of pro-inflammatory cytokines (IL-1β, IL-6, TGF-β1) established potent immunomodulatory effects. STE extracts suppressed pro-apoptotic (Bax, Caspase-3, JNK) and upregulated anti-apoptotic (Bcl-2) genes, protecting hepatocytes. Histopathology confirmed STE, especially peel extract, mitigated APAP-induced necrosis in liver, brain, and heart.