Molecular biology reports · 2026
Mechanistic assessment of hepato-reproductive toxicity of Mentha pulegium L. essential oil in rats.
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Pattanashetti Laxmi A, Siddika S Ayesha, Swathi D S, Sumanth B, Mishra Manas, Kulkarni Goutami G, Hiremath Sushmita I, Patil Santosh B
Аннотация
BACKGROUND: Mentha pulegium L. essential oil (MPEO), or pennyroyal oil, is widely used in traditional medicine, yet its safety remains debated because of pulegone, a constituent linked to hepatotoxicity and reproductive toxicity. This study evaluated MPEO's hepato-reproductive toxicity using an integrated in-silico and in vivo approach.
METHODS: Phytochemical composition was determined by GC-MS, while network pharmacology and molecular docking were used to identify likely targets and pathways. Reproductive and developmental toxicity were then assessed in Wistar rats (OECD Guideline 421) given 100, 200, or 400 mg/kg MPEO during premating, mating, gestation, and lactation, evaluating reproductive performance, developmental outcomes, biochemical markers, oxidative stress, hormone levels, and histopathology.
RESULTS: GC-MS confirmed pulegone (42.72%) as the major constituent, and network pharmacology revealed 116 shared targets, with CYP19A1, ESR2, and HSD11B1 emerging as key hub genes in steroid hormone biosynthesis and estrogen signaling. Molecular docking showed favorable pulegone binding affinities for these targets (-5.9, -6.0, and - 6.6 kcal/mol, respectively). In vivo, MPEO exposure produced dose-dependent impairments in fertility, implantation, litter size, pup survival, and offspring growth, alongside elevated liver injury markers, oxidative stress, hormonal disruption, and histopathological damage in liver, testes, and ovaries.
CONCLUSION: Pulegone-rich MPEO induces hepato-reproductive toxicity through endocrine disruption, oxidative tissue injury, and downregulation of steroid hormone biosynthesis signaling pathway. The novelty of this study lies in establishing the first multi-scale mechanistic link between computational targets (CYP19A1, ESR2, HSD11B1) and in vivo toxicity outcomes, offering insight for toxicology risk assessment and regulatory limits on pulegone-containing phytoactives.
METHODS: Phytochemical composition was determined by GC-MS, while network pharmacology and molecular docking were used to identify likely targets and pathways. Reproductive and developmental toxicity were then assessed in Wistar rats (OECD Guideline 421) given 100, 200, or 400 mg/kg MPEO during premating, mating, gestation, and lactation, evaluating reproductive performance, developmental outcomes, biochemical markers, oxidative stress, hormone levels, and histopathology.
RESULTS: GC-MS confirmed pulegone (42.72%) as the major constituent, and network pharmacology revealed 116 shared targets, with CYP19A1, ESR2, and HSD11B1 emerging as key hub genes in steroid hormone biosynthesis and estrogen signaling. Molecular docking showed favorable pulegone binding affinities for these targets (-5.9, -6.0, and - 6.6 kcal/mol, respectively). In vivo, MPEO exposure produced dose-dependent impairments in fertility, implantation, litter size, pup survival, and offspring growth, alongside elevated liver injury markers, oxidative stress, hormonal disruption, and histopathological damage in liver, testes, and ovaries.
CONCLUSION: Pulegone-rich MPEO induces hepato-reproductive toxicity through endocrine disruption, oxidative tissue injury, and downregulation of steroid hormone biosynthesis signaling pathway. The novelty of this study lies in establishing the first multi-scale mechanistic link between computational targets (CYP19A1, ESR2, HSD11B1) and in vivo toxicity outcomes, offering insight for toxicology risk assessment and regulatory limits on pulegone-containing phytoactives.