Hasawi lemon, locally termed "Hasawi Lomi," is a regional variety of Citrus aurantiifolia. Cultivated predominantly in the Al-Ahsa region of Saudi Arabia, this variety is well-known for its unique flavor, aromatic properties, and high acidity. There is a deficiency of scientific studies specifically focusing on the phytochemical composition and medicinal properties of the Hasawi lemon. Given the distinct environmental conditions of Al-Ahsa, it is anticipated that the Hasawi lemon could exhibit a unique phytochemical profile. Therefore, targeted phytochemical analyses and pharmacological assessments specific to the Hasawi lemon are essential to fully elucidate its medicinal potential and explore its potential applications in pharmaceutical and nutraceutical products.

The genus Citrus, family Rutaceae, comprise of 17 species, and the commonly cultivated are lemons (Citrus lemon), tangerines/mandarin (C. reticulata), oranges (C. sinensis), and limes (C. aurantifolia) (Brah et al., 2023). Ethnopharmacological records describe C. aurantifolia is widely used in traditional systems for its antitoxic, stomachic, and appetite-enhancing properties (Arias and Ramón-Laca, 2005). Ancient and medieval sources, particularly from Arabic and Mediterranean traditions, reported lime as antidotes against poison and tonics for digestion. Modern ethnopharmacological studies have identified flavonoids, saponins, and essential oils in lime with antibacterial and immunomodulatory effects. Moreover, its traditional use as a diuretic and antitoxic agent supports its ethnomedicinal relevance in managing renal ailments such as urolithiasis and toxin-induced nephropathies (Rahayu et al., 2024).

Lime oil possess various biological properties including anti-inflammatory (Amorim et al., 2016), antioxidant, hypolipidemic (Lin et al., 2019), antiulcer (Studiawan et al., 2023), anti-cancer (Narang and Jiraungkoorskul, 2016), Antiosteoporotic (Shalaby et al., 2011), antimicrobial (Al-Aamri et al., 2018), antityphoid and hepatoprotective properties (Jain et al., 2020). Because of the effective antibacterial and antifungal actions, citrus oil is used in skin care products (Jain et al., 2020) and in agriculture and food industry (Jain et al., 2020). Furthermore, C. aurantifolia essential oil with ketotifen administration inhibited weight gain via promoting anorexia (Asnaashari et al., 2010). C. aurantifolia essential oils inhibited AChE and BChE anti-cholinesterase effect (Tundis et al., 2012). CAEO displayed glucose lowering action and ameliorated hyperglycemia related dyslipidemia complications in alloxanized animals (Ibrahim et al., 2019). Regarding the hepatic actions, C. aurantifolia leaf extract protect against acetaminophen provoked hepatic injury (Adanma et al., 2019). While hydroethanol extracts of C. aurantifolia fruits protect against CCl4-induced hepatotoxic rats (Okoro, 2019). Concerning the renal effects, Citrus aurantifolia peel extract diminished nephrotoxicity prompted via doxorubicin (Oyinloye, E.O. et al., 2024) or by gentamicin (Oyinloye, E. et al., 2024). Moreover, polyphenol-rich Citrus leaf extract reduced renal toxicity caused by feeding with heated palm oil (Li et al., 2019).

Methotrexate (MTX) is used in several types of carcinomas as well as autoimmune syndromes, (Salem et al., 2017). MTX and MTX-polyglutamate (MTX-PGs) deter dihydrofolate reductase, making DNA replication unachievable thus averts cell growth. Accumulation of MTX-PGs leads to liver enzyme levels escalation, causing MTX prompted hepatic injury (Schmidt et al., 2022). Renal excretion is the primary route of elimination for methotrexate (MTX) and its metabolites. Patients treated with a high-dose MTX may develop acute kidney injury (AKI). MTX and its metabolite, 7-hydroxymethotrexate (7-OH-MTX) precipitate in renal tubules causing AKI in acidic conditions (Ali et al., 2024). Numerous reports stated MTX side effects, include gastrointestinal disorders, intestinal mucositis (Ali et al., 2024; Zhou et al., 2018), myelosuppression (Manappallil et al., 2018), kidney damage, reproductive toxicity (Felemban et al., 2020), and life-threatening liver dysfunction (Ali et al., 2017; Bu et al., 2018). MTX toxicity comprises of oxidative stress, inflammation, and destruction of cells leading to organ injury and damage (Widemann and Adamson, 2006). There are increasing interest in using natural plant-based therapies to counteract MTX associated toxicities. For instance, curcumin (Salem et al., 2017), Vitamin C (Savran et al., 2017), Ginkgo biloba (Sherif and Al-Shaalan, 2022), Chlorogenic acid (Ali et al., 2017) and Ellagic acid (Ebrahimi et al., 2019) among others.

The chemoprotective potential of C. aurantifolia fruit peel essential oil (Hasawi lemon) (CAEO) against MTX-induced toxicities has not been investigated earlier. Therefore, the aim of this study was to clarify the molecular mechanisms involved in MTX-induced renal and hepatic injuries and to explore the potential protective effects of C. aurantifolia fruit peel essential oil in these pathological contexts. Specifically, to assess the roles of Janus kinase/signal transducer and activator of transcription 3/nuclear factor-kappa B (JAK2/STAT3/NF-κB) signaling pathway, as well as the nuclear factor erythroid 2–related factor 2/heme oxygenase-1/NAD(P)H quinone dehydrogenase 1 (Nrf2/HO-1/NQO1) axis, in MTX-induced hepatorenal injuries.