Pesticides are one of the most widely used tools in pest management globally, both in agriculture and in domestic settings (Kotia et al., 2025). In particular, organophosphates (OPs) and pyrethroids are among the most widely used classes in terms of effects, breadth of use and commercial availability (Bayrak et al., 2025; Cavallaro et al., 2023; Ramya et al., 2023; Roy et al., 2026; Tudi et al., 2021). However, the increasing intensification of industrial agriculture and the extensive use of chemical formulations, often associated with repeated treatments and mixtures of active ingredients, have increased the likelihood of release into the environment and accidental or chronic exposure of non-target organisms (Barathinivas et al., 2022; Hossinian et al., 2025; Sula et al., 2020; Stara et al., 2019; Vajargah et al., 2021).

Among these is quinolphos (O, O-diethyl O-quinoxalin-2-yl phosphorothioate), a versatile organophosphate insecticide, which has been widely used for crop protection due to its effectiveness against a diverse array of pests (Gupta et al., 2017). Its pervasive utilization has engendered considerable apprehensions regarding detrimental health implications for non-target organisms, including humans (Hernández et al., 2013; Ismail et al., 2026; Mostafalou and Abdollahi, 2016). In general OPs, including quinolphos, are known for their detrimental impact on liver and kidney function (Saha et al., 2025).

Pesticide exposure through occupational contact, aerial drift, and inadequate protective measures represents a major health concern for agricultural workers and nearby populations, contributing to hepatorenal damage and chronic diseases (Damalas and Koutroubas, 2016; Jayasumana et al., 2015; Lebov et al., 2015). Consequently, there is an urgent need to develop efficient detoxification approaches employing natural protective agents.

The liver and kidneys, key organs for xenobiotic biotransformation and elimination, are particularly vulnerable to pesticide-induced injury (Abdel-Daim et al., 2018; Jindal et al., 2024; Kalender et al., 2012; Mousaviyon et al., 2025). The liver serves as the primary site for OP metabolism, where bioactivation pathways can generate reactive intermediates and electrophilic metabolites that disrupt cellular homeostasis and promote hepatocellular damage (Abdel Moneim, 2016). In parallel, the kidneys, through their high perfusion rate and filtration function, may accumulate parent compounds and metabolites, making renal tissue especially prone to both acute and chronic pesticide-related nephrotoxicity (El-Demerdash, 2011; Mansour and Mossa, 2010). Quinolophos toxicity has been associated with excessive reactive oxygen species (ROS) generation and oxidative stress, leading to lipid peroxidation, mitochondrial dysfunction, and depletion or inhibition of endogenous antioxidant defences (Chromcova et al., 2015; Padmanabha et al., 2015). These interrelated events can trigger membrane destabilization, impaired energy metabolism, and activation of inflammatory cascades, ultimately culminating in structural and functional alterations such as cellular degeneration, inflammatory infiltration, necrosis, and progressive fibrotic remodeling in both hepatic and renal tissues (Padmanabha et al., 2015).

Due to the limitations and adverse effects of synthetic pharmacological agents, there is growing interest in researching natural alternatives that can alleviate the toxicity induced by contaminants (Akinwande et al., 2019; Multisanti et al., 2025a; Impellitteri et al., 2025). These include substances derived from plant sources, including plants (Multisanti et al., 2025b; Impellitteri et al., 2025). Plants belonging to the genus Ziziphus have been recognised in traditional medicine for their various pharmacological properties, such as antioxidant, anti-inflammatory, hepatoprotective, and nephroprotective effects (Pawlowska et al., 2009). The plant Z. nummularia, which is used in traditional medicine, has gained attention because of its alleged nephroprotective and hepatoprotective properties (Mesmar et al., 2022). Z. nummularia, referred to as wild jujube, is a xerophytic shrub indigenous to arid and semi-arid areas of Asia and Africa. Phytochemical analyses have revealed bioactive compounds in Z. nummularia leaves, such as flavonoids, saponins, alkaloids, tannins, and phenolic compounds, which collectively enhance its pharmacological potential (Mesmar et al., 2022). Ethnobotanical surveys document its traditional use in South Asian folk medicine for treating fever, inflammation, gastrointestinal disorders, and pain, with leaves and bark particularly valued for hepatoprotective properties (Khare, 2011; Abbasi et al., 2012).

Although various Ziziphus species have demonstrated hepatoprotective and nephroprotective effects, information on Z. nummularia’s protective effects against organophosphate-induced toxicity remains limited. Previous studies have primarily focused on biochemical parameters, with fewer incorporating comprehensive histopathological and behavioral assessments (Farkhondeh et al., 2024; Yu et al., 2021).

This study systematically investigates the hepatoprotective and nephroprotective properties of Z. nummularia leaf extract against quinalphos-induced toxicity in Wistar rats. Using an integrative approach combining standardized histopathological lesion scoring (0–3 scale) with behavioral evaluations, we aim to establish the dose-dependent protective effects of the extract and elucidate the relationship between microscopic tissue changes and functional outcomes.

The study examines the protective effects of Z. nummularia leaf extract against quinolphos-induced toxicity in Wistar rats, providing valuable insights into its potential pharmacological applications for alleviating pesticide-induced organ damage. The results of the study reveal significant changes in behavioural patterns and histopathological structures, supporting the protective function of the extract. During the experimental period, a significant disparity was observed in the behaviour of the control group compared to the quinolphos-exposed rats. The control group exhibited normal activity levels, characterised by alertness and active movement, whereas the quinolphos exposure group demonstrated notable behavioural abnormalities, including diminished locomotor activity, as indicated by a rough coat texture and substantial body weight loss. The noticeable reduction in activity and coarse coat texture suggests systemic toxicity linked to cholinesterase inhibition from organophosphate exposure (Salvi et al., 2003). The observed behavioural deficits correspond with other research indicating that exposure to agrochemicals, including organophosphates, causes systemic toxicity, leading to discernible behavioural abnormalities in rodent models (Badr, 2020). These behavioural alterations align with established effects of organophosphate pesticides that impair central nervous system function, leading to reduced motor skills and cognitive deficits (Chen et al., 2021). The decline in mobility and grooming within the quinolphos group corresponds with documented toxicological effects of organophosphate exposure (Badr, 2020).

The study also emphasised a dose-dependent response to the Z. nummularia extract. The behavioural recovery noted in the high-dose Z. nummularia extract group (500 mg/kg) indicates a substantial dose-dependent pharmacological potential, suggesting that this extract may mitigate the neurotoxic effects of quinolphos. The high-dose treatment group exhibited significant improvements, evidenced by recovery from anorexia and enhanced grooming behaviours, corroborating the hypothesis that elevated doses of natural extracts frequently augment protective effects against induced toxicity (You et al., 2025). The enhancement in appetite and overall clinical status in the high-dose treatment group signifies successful detoxification and reestablishment of physiological homeostasis, aligning with research that points out the hepatoprotective properties of diverse medicinal plant extracts (Olukanni et al., 2025). The phytochemical characteristics of the Ziziphus extract, which are known to contain bioactive compounds with potential antioxidant activity, may be partially responsible for the behavioural recovery seen when the extract was administered (Mesmar et al., 2022; Ray et al., 2024). This underscores the importance of Z. nummularia in traditional medicine, especially regarding its application as a neuroprotective agent (Boutlelis et al., 2025).

In the low-dose treatment group (250 mg/kg), partial symptom alleviation was noted; however, persistent indications of diminished locomotor activity and fatigue suggest that an increased dosage may be required for maximal effects. This corresponds with findings highlighting the importance of concentration in deploying plant extracts as antidotes against chemical-induced toxicity (Rongzhu et al., 2007). The observed behavioural enhancements correspond with changes in biochemical parameters, underscoring the protective properties of specific plant extracts against oxidative stress and inflammation (Liu et al., 2011).

Furthermore, the association between behavioural enhancement and histopathological observations indicates that Z. nummularia extract may operate on various levels to alleviate quinolphos-induced damage. Z. nummularia has become known for its bioactive compounds possessing antioxidant properties, potentially mitigating oxidative stress and inflammation caused by organophosphates (Dickey et al., 2007). This highlights the crucial function of natural extracts in safeguarding and revitalising impaired organ systems. The observed survival rate among treatment groups indicates that Z. nummularia administration may improve resistance to organophosphate toxicity. Moreover, specific Ziziphus species have demonstrated sedative, analgesic, anxiolytic, and anti-nociceptive effects, indicating their role in regulating central nervous system activity (Henneh et al., 2021; Kumar et al., 2023; Chung et al., 2025). The incorporation of traditional herbal remedies such as Z. nummularia into modern treatment protocols shows potential for mitigating organophosphate-induced toxicity, due to their natural composition and reduced side effects relative to synthetic pharmaceuticals (Mesmar et al., 2022; Imran et al., 2023).

The histopathological examinations indicate substantial insights into the structural damage caused to renal tissues by quinolphos exposure. The control and vehicle-treated groups exhibited no discernible pathological changes, in contrast to the quinolphos-treated group, which displayed significant alterations in multiple renal compartments. The results reveal significant glomerular cell debris, marked basement membrane irregularities, tubular injury, and interstitial oedema, suggesting considerable renal dysfunction due to the organophosphate toxin. These observations correspond with existing literature that records the nephrotoxic effects of organophosphates and their ability to induce acute kidney injuries characterised by glomerular and tubular disruptions, as noted in previous studies on various nephrotoxins (Sobolev et al., 2022).

The administration of Z. nummularia leaf extract exhibited significant nephroprotective effects that were dependent on dosage. The high-dose treatment group exhibited nearly complete recovery, reinforcing the hypothesis that phytochemical metabolites in Z. nummularia positively influence renal architecture and function. Prior research has validated these results, indicating that extracts from Ziziphus species exhibit antioxidant and anti-inflammatory properties that alleviate nephrotoxicity caused by various agents (Mesmar et al., 2022). The low-dose group demonstrated only minor histopathological alterations, further suggesting the extract’s potential to mitigate quinolphos-induced nephrotoxicity, likely due to its phytochemicalss, including flavonoids and saponins, recognised for their protective properties on renal tissues (Mesmar et al., 2021).

The noticeable improvement in kidney histopathology noted in the high-dose group corresponds with the increasing evidence indicating that natural extracts may serve as potent pharmacological agents against nephrotoxicity. Animal models administered different plant extracts have demonstrated a restoration of renal architecture and function, indicating their potential to mitigate the harmful effects induced by nephrotoxic agents (Jeshan et al., 2021). These findings suggest that Z. nummularia is a promising alternative for further pharmacological investigation and clinical use in renal protection against toxic environmental agents.

The microscopic examination of liver sections has provided essential insights into the hepatic histopathological changes caused by quinolphos. The studies indicate significant hepatotoxic alterations accompanied by profound changes in multiple hepatic parameters. The quinolphos-induced group exhibited significant abnormalities, including pronounced alterations in hepatocyte morphology, disturbances in the hepatic portal veins and triad structures, extensive leukocyte infiltration, sinusoidal congestion, and hepatocyte necrosis, all assessed with considerable severity. Furthermore, moderate pathological alterations were noted in additional parameters, encompassing changes in central vein structure, cellular lysis, bile duct pathology, vascular degeneration, cellular swelling, vacuolization, and localised leukocyte infiltration. The control group and vehicle group exhibited no histopathological abnormalities, highlighting the harmful effects of quinolphos.

The administration of Z. nummularia leaf extract exhibited substantial hepatoprotective effects at both low (250 mg/kg) and high doses (500 mg/kg). In the low dose group, moderate hepatocyte alterations, hepatic portal vein changes, and leukocyte infiltration were evident, indicating persistent hepatotoxicity. However, even in this group, improvements were observed in a number of evaluated parameters, such as hepatocyte necrosis, which was noticeably less severe than in the quinolphos group. The elevated dosage demonstrated enhanced protective effects, evidenced by a significant decrease in hepatocyte alterations and total resolution of vascular degeneration, thereby indicating a considerable restoration of hepatic vascular integrity, suggesting the intricate role of phytochemicals in mitigating chemically induced liver pathology (Mesmar et al., 2022).

This synergy indicates that Z. nummularia contains bioactive compounds that can mitigate oxidative stress and inflammation associated with hepatotoxicity, thereby supporting its traditional application in liver disorders. Multiple studies on Z. nummularia highlight its abundant phytochemical composition, encompassing alkaloids and flavonoids, which enhance its pharmacological advantages (Abdulrahman et al., 2022; Mesmar et al., 2022).

The present study confirmed our hypothesis that Z. nummularia leaf extract provides dose-dependent protection against quinalphos-induced hepato-renal toxicity. This hypothesis was substantiated by dose-dependent behavioral recovery (500 mg/kg > 250 mg/kg), near-complete histopathological resolution at high dose versus partial protection at low dose. The vehicle control exhibited no adverse effects, validating the non-toxic nature of the extract.

This study confirms our hypothesis that Z. nummularia leaf extract provides dose-dependent protection against quinalphos-induced hepato-renal toxicity. The integrative method that incorporates behavioural monitoring with comprehensive histopathological grading effectively revealed that quinalphos exposure causes significant multi-organ damage, characterised by prominent nephrotoxic and hepatotoxic changes, such as glomerular atrophy, tubular degeneration, interstitial oedema, hepatocyte necrosis, and sinusoidal congestion. The high-dose Z. nummularia treatment (500 mg/kg) resulted in nearly complete histopathological recovery in renal and hepatic tissues, with complete disappearance of most pathological parameters and restoration of vascular integrity, while the low-dose treatment (250 mg/kg) provided partial protection. The vehicle control group exhibited no adverse effects, confirming the safety profile of Z. nummularia extract. The identified protective mechanisms probably relate to the abundant phytochemical composition of Z. nummularia, especially flavonoids, saponins, and phenolic compounds with antioxidant and anti-inflammatory attributes that mitigate organophosphate-induced oxidative stress. The relationship between behavioural recovery and histopathological restoration indicates the systemic protective effects of the extract. These findings establish Z. nummularia as an appropriate option for development as a natural prophylactic or pharmacological agent for populations at risk of organophosphate exposure, particularly agricultural workers in areas with high pesticide usage, and necessitate further mechanistic investigations and clinical translation studies.