JAHP_14_1

Introduction

The kidneys serve as essential regulators of systemic homeostasis which performing critical roles in fluid and electrolyte balance, waste product excretion, acid–base regulation and endocrine-signaling (Crintea et al., 2025). These vital organs are particularly susceptible to damage from a range of toxic-agents including xenobiotics, pharmaceuticals and environmental compounds (Iovdijová and Bencko, 2010). In companion animals, kidney toxicity can result from various causes such as drugs, environmental toxins, and other harmful substances, and may ultimately lead to death. In humans, ethanol is a widely consumed substance known to induce systemic toxicity when ingested in high quantities (Le Dare et al., 2019). Excessive ethanol exposure can lead to both acute and chronic renal impairment primarily through mechanisms involving oxidative-stress, inflammation and direct cytotoxic effects on renal tissues (Rodrigo and Rivera, 2002). Ethanol metabolism generates excessive reactive oxygen species (ROS) which disrupt the cellular redox state and overwhelm intrinsic antioxidant defenses (Contreras-Zentella et al., 2022). This oxidative burden coupled with ethanol-induced upregulation of pro-inflammatory cytokines contributes to functional deterioration and structural damage in nephron units particularly in glomerular and tubular-components (Yang et al., 2022).

Early diagnosis of renal-dysfunction is essential for preventing the progression of irreversible kidney damage (Locatelli et al., 2002). Traditionally, serum creatinine and urea are employed to assess renal-function. However, these conventional markers are influenced by numerous confounding variables such as age, body mass, dietary protein intake and hydration-status (Al-Ziaydi et al., 2020). These limitations hinder their sensitivity, especially in detecting mild or subclinical renal impairment. In recent years, cystatin C has emerged as a more accurate and sensitive biomarker for assessing glomerular-filtration-rate (GFR). Cystatin C is a low-molecular-weight cysteine protease inhibitor produced consistently by all nucleated cells. It is freely filtered by the glomeruli and almost completely reabsorbed and degraded in the renal tubules without significant-secretion (Onopiuk et al., 2015). Significantly, its serum concentration is unaffected by age, sex, muscle mass or diet making it a more reliable index of GFR than creatinine (Rajagopalan et al., 2016). Elevated serum cystatin C levels are indicative of early-stage renal-dysfunction and have also been linked to systemic-inflammatory-responses, cardiovascular-pathology and metabolic abnormalities (Tanwar et al., 2021). Therefore, cystatin C holds promise not only as a renal function biomarker but also as a broader indicator of systemic stress and disease.

Numerous medicinal plants are known to possess potent antioxidant and anti-inflammatory properties which may contribute to cellular protection in organ systems vulnerable to oxidative and inflammatory-damage (Ondua et al., 2019). E. sativav generally referred to as (rocket or arugula) is one such plant that has attracted considerable attention. It belongs to the Brassicaceae family and has long been consumed for both culinary and traditional medicinal purposes (Barazani and Ziffer-Berger, 2014). Rich in a diverse array of phytochemicals including glucosinolates, isothiocyanates, flavonoids, phenolic acids, vitamins and minerals. E. sativa exhibits a wide spectrum of pharmacological activities (Jin et al., 2009). These include antioxidative, anti-inflammatory, antimicrobial, hepatoprotective, cardioprotective and anticancer effects (Duru et al., 2022).

The antioxidative properties of E. sativa are primarily attributed to its capacity to scavenge ROS enhance endogenous-antioxidant enzymes such as superoxide dismutase and glutathione-peroxidase and reduce lipid peroxidation (Piragine, 2020). Additionally, the plant exerts anti-inflammatory effects through downregulation of key proinflammatory mediators, including tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) (Ochar et al., 2024). Its isothiocyanate content particularly erucin derived from glucoerucin which is believed to modulate nuclear factor kappa B (NF-κB) signaling and other molecular pathways involved in inflammation (Genah et al., 2024). These pharmacodynamic properties suggest a promising role for E. sativa in counteracting ethanol-induced toxicity which is known to be mediated by similar oxidative and inflammatory mechanisms (Vicidomini et al., 2024).

Ethanol-induced kidney injury is typically characterized by structural alterations in the renal cortex and medulla including glomerular atrophy, tubular epithelial degeneration and interstitial infiltration by inflammatory cells (Rikalo and Romanenko, 2018). These histopathological features are accompanied by biochemical derangements, including elevated serum urea and creatinine, as well as increased levels of oxidative stress biomarkers such as malondialdehyde (Dinu et al., 2006). The depletion of intracellular antioxidants further exacerbates renal damage (Dennis and Witting, 2017). In this context, therapeutic agents capable of restoring antioxidant balance and attenuating inflammation may preserve renal architecture and function (Lee et al., 2024). E. sativa due to its phytochemical constituents, is hypothesized to confer such benefits. This study aims to evaluate the effects of E. sativa leaf extract on serum cystatin C levels and overall renal function in a rat model of ethanol-induced nephrotoxicity.

Materials and Methods

Plant material and extract preparation

Fresh E. sativa leaves were procured from a local source and authenticated by a qualified botanist. The leaves were thoroughly rinsed with distilled water then air-dried under shade at “ambient room temperature” for seven days. Subsequently, the dried leaves were pulverized into a fine powder using an electric-grinder. A quantity of 100 grams of the powdered material was subjected to maceration in 70% ethanol at a ratio of 1:10 (w/v) for 72 hours with intermittent agitation. The resulting extract was filtered through muslin cloth followed by Whatman No.1 filter paper (Abed, 2015; Ali et al., 2024). The filtrate was concentrated under reduced pressure at 40°C using a rotary evaporator to yield a semi-solid-extract, which was stored at 4°C in airtight containers until further use.

Experimental animals and design

Thirty male Wistar rats, each weighing between 210 and 275 grams, were obtained and allowed to acclimate for one week under controlled laboratory conditions temperature maintained at 22 ± 2°C, relative humidity at 55 ± 5%, and a 12-hour light/dark cycle. Animals had unrestricted access to standard laboratory chow and water throughout the study. The experimental protocol received prior approval from The experimental protocol received prior approval from the Animal Ethical Research committee, college of dentistry, Al-Iraqia university (Ethical approval Number: ESA and HER- 10-10-07-2025), ensuring compliance with accepted guidelines for the care and use of laboratory animals (Kiani et al., 2022).

The ethanol (95% ethyl alcohol, BDH England) used in the current was purchased from local medical laboratory supplies sales office, Baghdad, Iraq. Ethanol was used at concentration of 60% v/v diluted by distilled water. A 5 g/kg/day was the dose of ethanol at estimated volume between 2.3 to 2.9 ml for each rat.

The animals were randomly allocated into five groups of six rats each (n = 6) when group I (Normal-Control) received distilled water orally and served as the untreated control group, group II (Ethanol Control) administered ethanol orally at a dose of 5 g/kg/day to induce renal toxicity according to previously published study (Pruett et al., 2020). Group III (Ethanol + E. sativa 250 mg/kg) treated with ethanol plus E. sativa extract at 250 mg/kg/day. Group IV (Ethanol + E. sativa 375 mg/kg) treated with ethanol plus E. sativa extract at 375 mg/kg/day. Finally, group V (Ethanol + E. sativa 500 mg/kg) was treated with ethanol plus E. sativa extract at 500 mg/kg/day.

All interventions were administered orally via gavage once daily for a total duration of 30 days. In the treatment groups, E. sativa extract was administered one hour prior to ethanol to assess its potential nephroprotective effects.

Sample collection and biochemical measurements

At the end of the experimental period, animals were fasted overnight and anesthetized with intraperitoneal injections of ketamine (80 mg/kg) and xylazine (10 mg/kg). Blood samples were collected by cardiac-puncture, allowed to clot at room temperature, and then centrifuged at 3000 revolutions per minute for 15 minutes (Abed et al., 2023). Serum was carefully separated and stored at −20°C pending biochemical analysis.

Serum cystatin C concentrations were quantified using a species-specific sandwich enzyme-linked immunosorbent assay (ELISA) kit (Shanghai DeBo Biochemistry Laboratory Inc). Then, the absorbance of the samples was measured at 450 nm with ELISA Lab immunoassay reader (Cobrin et al., 2013). The results were displayed as µg/ mL Cystain C in the blood. The creatinine serum detection kit was used to estimate the level of creatinine using colorimetric measurement method and according to instruction of the manufacture (StressXpress®). Additionally, the urea was also estimated by colorimetric assay kit (Urease method) (E-BC-K183-S) and measured by spectrophotometer (at wave length of 580 nm) according to the instruction of the manufacture.

Statistical analysis

Data were expressed as mean values with corresponding standard-deviations (mean ± SD). Statistical comparisons among groups were performed using one-way analysis of variance (ANOVA), followed by “Tukey’s post hoc test” for multiple comparisons. Differences were considered statistically-significant when p-values were less than 0.05 (Abed et al., 2022). Statistical-analyses and graphical representations were carried out using GraphPad-Prism software (version 6).

Results and Discussion

The present results assessed the renoprotective potential of E. sativa leaf extract against ethanol-induced nephrotoxicity by evaluating serum concentrations of cystatin C, urea and creatinine. Chronic ethanol administration (5 g/kg/day for 30 days) elicited significant renal-impairment, evidenced by marked elevations in all measured biomarkers compared to the normal control group (p ≤ 0.05), the results are illustrated in Figures 1-3.

Serum cystatin C, a highly sensitive marker for early renal dysfunction and decline in glomerular filtration rate, increased significantly from 0.52 ± 0.05 mg/L in the control group to 1.25 ± 0.12 mg/L in ethanol-treated rats (p ≤ 0.05). This substantial increase indicates significant glomerular injury induced by ethanol toxicity, confirming the utility of cystatin C as a reliable early biomarker in nephrotoxicity models. Importantly, treatment with E. sativa extract elicited a statistically-significant, dose-dependent reduction in cystatin C levels: 0.98 ± 0.09 mg/L, 0.75 ± 0.07 mg/L, and 0.58 ± 0.06 mg/L at 250, 375, and 500 mg/kg doses, respectively (p ≤ 0.05 versus ethanol group). The near normalization of cystatin C at the highest dose underscores the extract’s potent capacity to mitigate early glomerular damage as in Figure 1. Cystatin C is a low molecular weight protein that produced consistently by nucleated cells. It is filtrated freely at the glomerulus and reabsorbed and catabolized by cell of the proximal tubule (Abrahamson et al., 1990). The result of the current study showed that, the obvious decreasing of cystatin C level in groups treated with E. sative suggests that the plants components including the flavonoids, glucosinolates, and isothiocyanates have a protection effects and restore the glomerular filtration capacity by ameliorative oxidative and inflammatory insults caused by ethanol metabolism. These results agree with previously published study (Sarwar et al., 2007), who approved the antioxidant and protective activities of E. sative seeds on the mercuric chloride induced renal toxicity.

Serum urea is an indicator of renal nitrogenous waste clearance, rose significantly from 5.3 ± 0.6 mmol/L in the control group to 12.8 ± 1.2 mmol/L following ethanol exposure (p ≤ 0.05). Administration of E. sativa resulted in significant, dose-dependent decreases in serum urea: 9.5 ± 0.9 mmol/L, 7.2 ± 0.7 mmol/L, and 6.1 ± 0.5 mmol/L at the respective doses (p ≤ 0.05 vs. ethanol group). These reductions suggest restoration of renal excretory function and amelioration of tubular impairment caused by ethanol (Figure 2). A treatment with E. sativa extract led to progressive and dose dependent decreases in urea concentration that approach the normal values at higher doses. The decrease in urea levels suggests an improvement in nitrogen clearance, likely due to the regeneration of tubular structures and enhanced renal perfusion. This finding is consistent with previously published studies (Hannan et al., 2021), which demonstrated the pharmacological activity of Nigella sativa and its bioactive constituent, thymoquinone, in protecting against kidney injury.

Serum creatinine, a conventional marker of glomerular filtration, was also significantly elevated by ethanol treatment, increasing from 35.4 ± 3.1 µmol/L in the control group to 68.7 ± 5.4 µmol/L (p ≤ 0.05). Treatment with E. sativa extract yielded significant dose-dependent decreases in creatinine concentration: 52.3 ± 4.7 µmol/L, 42.8 ± 3.9 µmol/L, and 37.1 ± 3.4 µmol/L for 250, 375, and 500 mg/kg doses, respectively (p ≤ 0.05 compared to ethanol-only group). These results indicate improved glomerular filtration capacity and reduced renal dysfunction (Figure 3). The level of serum creatinine is extensively used in clinical laboratory investigation. However, it is less sensitive to early kidney dysfunction because it elevated only after substantial nephron loss (Ávila et al., 2025). These results are compatible with previously published study by (Boukarine et al., 2023), who approved the antioxidant therapeutic effects of E. sativa against hepato-renal toxicity caused by xylene in Wistar rats.

The nephroprotective effects observed are likely attributable to the phytochemical constituents of E. sativa, which include flavonoids, glucosinolates, and phenolic compounds with well-documented antioxidant and anti-inflammatory properties (Raquel and Fernando, 2016). These bioactive molecules mitigate oxidative stress by scavenging reactive oxygen species generated during ethanol metabolism, thereby preventing lipid peroxidation and cellular damage in renal tissues (Sharma et al., 2023). Furthermore, the extract may inhibit proinflammatory cytokine release (e.g., TNF-α, IL-6), thereby attenuating inflammation-mediated renal injury (Thiruvengadam et al., 2024). The cumulative antioxidant and anti-inflammatory activities of E. sativa plausibly underpin the preservation of renal structural and functional integrity, as reflected by normalized serum biomarkers (Fuentes et al., 2014).

The dose-dependent improvements across cystatin C, urea, and creatinine not only confirm the extract’s efficacy but also emphasize the importance of appropriate dosing to maximize therapeutic benefits. Of particular note is the significant reduction in cystatin C, which highlights the extract’s capability to prevent early glomerular filtration rate decline a critical advantage for early intervention strategies (Seronie-Vivien et al., 2008). These results are consistent with existing studies on E. sativa’s protective effects in models of organ toxicity and extend its application to renal protection in ethanol-induced nephrotoxicity (Alam et al., 2007; Boukarine et al., 2023).

Conclusion

E. sativa leaf extract significantly attenuates ethanol-induced renal dysfunction in rats, as demonstrated by the dose-dependent reduction in serum cystatin C, urea, and creatinine levels. These results suggest that E. sativa possesses potent antioxidant and anti-inflammatory properties capable of preserving glomerular and tubular function. This study provides a scientific rationale of E. sativa as a natural therapeutic agent for protecting renal health against alcohol-related toxicity.

Acknowledgement

The authors express their appreciation to technical staff for their assistance and collaboration throughout the course of this research. Special thanks are extended to the Department of basic science / College of Dentistry / Al-Iraqia universty for providing facilities and support that made this study possible.

Novelty Statement

Contrasting previous published studies, that focused on antioxidant and hepatoprotective effect of E. sativa, this study estimates the nephroprotective protentional using a sensitive biomarker (Cystain C) accompanying the traditional renal function markers. Moreover, this study gives a new vision into dose-response interactions and the possible therapeutic relevance of E. sativa in ameliorative renal dysfunction.

AUTHOR’S CONTRIBUTION

All authors contribute and collaborate equally in this article.

Funding

This is a self-funded research, no fund received from any authorities.

Ethical approval

This study has been approved by Ethical and research committee approval number: ESA and HER- 10-10-07-2025.

Data availability

All data generated or analyzed during the current study are available upon request.

Generative AI and AI-assisted technology statement

The authors declare that no Genrative AI was used in the creation of this manuscript.

Conflict of interest

All authors declared no conflict if interest.

References

Abed HH (2015). Study chemical composition of Borago officinalis L. extraction and antibacterial activity against gastrointestinal infections. Int. J., 3(6): 1530–1535.

Abed HH, Ali AM, Al-Ziaydi AG (2023). Evaluation level of serum vitamin B12 in Iraqi patients with diabetes mellitus type 2, who used the metformin drug as a hypoglycemic agent. Pak. J. Pharm. Sci., 36(2): 425–429.

Abed HH, Al-Ziaydi AG, Taher IA, Al-Dulaimi AK (2022). Comparison of some hematological parameters between male and female patients infected with COVID-19. Hum. Antibodies. Preprint, pp. 1–5. https://doi.org/10.3233/HAB-220006

Abrahamson M, Olafsson I, Palsdottir A, Ulvsbäck M, Lundwall Å, Jensson O, Grubb A (1990). Structure and expression of the human cystatin C gene. Biochem. J., 268(2): 287–294. https://doi.org/10.1016/0167-4781(90)90171-W

Alam MS, Kaur G, Jabbar Z, Javed K, Athar M (2007). Eruca sativa seeds possess antioxidant activity and exert a protective effect on mercuric chloride induced renal toxicity. Food Chem. Toxicol., 45(6): 910–920. https://doi.org/10.1016/j.fct.2006.11.013

Ali AM, Hussein NA, Jawad D, Abed HH, Mekkey SM (2024). Hepatotoxic effects of Stevia rebaudiana leaf extract and commercial stevia on rats: A comparative study. Rev. Clin. Pharmacol. Pharmacokinet., 38(2): 69–72. https://doi.org/10.61873/XFAQ5379

Al-Ziaydi AG, Abed HH, Ali AM, Mobder E, Nayif AHF, Fadhel AH (2020). Evaluate toxic effect of bisphenol a on kidney of male mice and mitigation its effect by green tea extract. J. Glob. Pharma Technol., 12(2): 664-670. https://jgpt.co.in/index.php/jgpt/article/view/3349/2646

Ávila M, Mora Sánchez MG, Bernal Amador AS, Paniagua R (2025). The metabolism of creatinine and its usefulness to evaluate kidney function and body composition in clinical practice. Biomolecules, 15(1): 41. https://doi.org/10.3390/biom15010041

Barazani O, Ziffer-Berger J (2014). Eruca sativa, a tasty salad herb with health-promoting properties. Med. Aromat. Plants Middle-East. pp. 269–279. https://doi.org/10.1007/978-94-017-9276-9_15

Boukarine R, Hamdi L, Khelili K (2023). Therapeutic effects of rocket leaves (Eruca sativa) against hepato-renal toxicity caused by xylene in Wistar rats. Pak. J. Zool., 55(3): 1437. https://doi.org/10.17582/journal.pjz/20211107181135

Cobrin AR, Blois SL, Kruth SA, Abrams-Ogg ACG, Dewey C (2013). Biomarkers in the assessment of acute and chronic kidney diseases in the dog and cat. J. Small Anim. Pract., 54(12): 647–655. https://doi.org/10.1111/jsap.12150

Contreras-Zentella ML, Villalobos-García D, Hernández-Muñoz R (2022). Ethanol metabolism in the liver, the induction of oxidant stress, and the antioxidant defense system. Antioxidants, 11(7): 1258. https://doi.org/10.3390/antiox11071258

Crintea IN, Cindrea AC, Mederle OA, Trebuian CI, Timar R (2025). Electrolyte imbalances and metabolic emergencies in obesity: Mechanisms and clinical implications. Diseases, 13(3): 69. https://doi.org/10.3390/diseases13030069

Dennis JM, Witting PK (2017). Protective role for antioxidants in acute kidney disease. Nutrients, 9(7): 718. https://doi.org/10.3390/nu9070718

Dinu D, Nechifor MT, Movileanu L (2006). Ethanol-induced alterations of the antioxidant defense system in rat kidney. J. Biochem. Mol. Toxicol., 19(6): 386–395. https://doi.org/10.1002/jbt.20101

Duru ÖA, Araç FÖÖ, Aldemir O (2022). The importance of arugula (Eruca sativa) and pharmacological effects of different phytochemical components in its content for human health. In: Handbook of research on advanced phytochemicals and plant-based drug discovery. IGI Global. pp. 327–340. https://doi.org/10.4018/978-1-6684-5129-8.ch016

Fuentes E, Alarcón M, Fuentes M, Carrasco G, Palomo I (2014). A novel role of Eruca sativa Mill. (rocket) extract: Antiplatelet (NF-κB inhibition) and antithrombotic activities. Nutrients, 6(12): 5839–5852. https://doi.org/10.3390/nu6125839

Genah S, Ciccone V, Filippelli A, Simonis V, Martelli A, Piragine E, Pagnotta E, Pecchioni N, Calderone V, Morbidelli L (2024). Erucin, a natural isothiocyanate, exerts pro-angiogenic properties in cultured endothelial cells and reverts angiogenic impairment induced by high glucose. Phytother. Res., 38(6): 2641–2655. https://doi.org/10.1002/ptr.8183

Hannan MA, Rahman MA, Sohag AAM, Uddin MJ, Dash R, Sikder MH, Rahman MS, Timalsina B, Munni YA, Sarker PP, Alam M, Mohibbullah M, Haque MN, Jahan I, Hossain MT, Afrin T, Rahman MM, Tahjib-Ul-arif M, Mitra S, Kim B (2021). Black cumin (Nigella sativa l.): A comprehensive review on phytochemistry, health benefits, molecular pharmacology, and safety. Nutrients, 13(6), Article 1784. https://doi.org/10.3390/nu13061784

Iovdijová A, Bencko V (2010). Potential risk of exposure to selected xenobiotic residues and their fate in the food chain-Part. I: Classification of xenobiotics. Ann. Agric. Environ. Med., 17(2): 183–192.

Jin J, Koroleva OA, Gibson T, Swanston J, Magan J, Zhang YAN, Rowland IR, Wagstaff C (2009). Analysis of phytochemical composition and chemoprotective capacity of rocket (Eruca sativa and Diplotaxis tenuifolia) leafy salad following cultivation in different environments. J. Agric. Food Chem., 57(12): 5227–5234. https://doi.org/10.1021/jf9002973

Kiani AK, Pheby D, Henehan G, Brown R, Sieving P, Sykora P, Marks R, Falsini B, Capodicasa N, Miertus S, Lorusso L, Dondossola D, Tartaglia GM, Ergoren MC, Dundar M, Michelini S, Malacarne D, Bonetti G, Dautaj A (2022). International bioethics study group. Ethical considerations regarding animal experimentation. J. Prevent. Med. Hyg., 63(2) (Suppl 3): E255–E266.

Le Dare B, Lagente V, Gicquel T (2019). Ethanol and its metabolites: Update on toxicity, benefits, and focus on immunomodulatory effects. Drug Metab. Rev. 51(4): 545–561. https://doi.org/10.1080/03602532.2019.1679169

Lee OYA, Wong ANN, Ho CY, Tse KW, Chan AZ, Leung GP-H, Kwan YW, Yeung MHY (2024). Potentials of natural antioxidants in reducing inflammation and oxidative stress in chronic kidney disease. Antioxidants, 13(6): 751. https://doi.org/10.3390/antiox13060751

Locatelli F, Vecchio LD, Pozzoni P (2002). The importance of early detection of chronic kidney disease. Nephrol. Dialysis Transplant., pp. 17. https://doi.org/10.1093/ndt/17.suppl_11.2

Ochar K, Iwar K, Nair VD, Chung YJ, Ha BK, Kim SH (2024). The potential of glucosinolates and their hydrolysis products as inhibitors of cytokine storms. Molecules, 29(20): 4826. https://doi.org/10.3390/molecules29204826

Ondua M, Njoya EM, Abdalla MA, McGaw LJ (2019). Anti-inflammatory and antioxidant properties of leaf extracts of eleven South African medicinal plants used traditionally to treat inflammation. J. Ethnopharmacol., 234: 27–35.

Onopiuk A, Tokarzewicz A, Gorodkiewicz E (2015). Cystatin C: A kidney function biomarker. Adv. Clin. Chem., 68: 57–69.

Piragine E (2020). Pharmacological modulation of intracellular signalling pathways involved in oxi-inflammageing and age-related cardiovascular pathologies by naturally occurring molecules.

Pruett S, Tan W, Howell GE, Nanduri B (2020). Dosage scaling of alcohol in binge exposure models in mice: An empirical assessment of the relationship between dose, alcohol exposure, and peak blood concentrations in humans and mice. Alcohol., 89: 9–17. https://doi.org/10.1016/j.alcohol.2020.03.011

Rajagopalan P, Abraham G, Reddy YNV, Lakshmanasami R, Prakash ML, Reddy YNV (2016). Population-based estimation of renal function in healthy young Indian adults based on body mass index and sex correlating renal volume, serum creatinine, and cystatin C. Int. J. Nephrol. Renovasc. Dis., pp. 243–247. https://doi.org/10.2147/IJNRD.S104983

Rikalo NA, Romanenko IV (2018). Morphological changes in kidney tissues of rats with acute ethanol-induced injury and after drug correction. J. Educ. Health Sport., 8(2): 272–279.

Raquel P.F. Guiné, Fernando J. Gonçalves, (2016). Bioactive compounds in some culinary aromatic herbs and their effects on human health. Mini Rev. Med. Chem., 16(11): 855–866. https://doi.org/10.2174/1389557516666160211120540

Rodrigo R, Rivera G (2002). Renal damage mediated by oxidative stress: A hypothesis of protective effects of red wine. Free Radic. Biol. Med., 33(3): 409–422. https://doi.org/10.1016/S0891-5849(02)00908-5

Sarwar AM, Kaur G, Jabbar Z, Javed K, Athar M (2007). Eruca sativa seeds possess antioxidant activity and exert a protective effect on mercuric chloride induced renal toxicity. Food Chem. Toxicol., 45(6): 910–920. https://doi.org/10.1016/j.fct.2006.11.013

Seronie-Vivien S, Delanaye P, Pieroni L, Mariat C, Froissart M, Cristol JP, Group S, Biology RF, RFW (2008). Cystatin C: Current position and future prospects. Clin. Chem. Lab. Med., 46(12): 1664–1686. https://doi.org/10.1515/CCLM.2008.336

Sharma P, Nandave M, Nandave D, Yadav S, Vargas-De-La-Cruz C, Singh S, Tandon R, Ramniwas S, Behl T (2023). Reactive oxygen species (ROS)-mediated oxidative stress in chronic liver diseases and its mitigation by medicinal plants. Am. J. Trans. Res., 15(11): 6321.

Tanwar V, Adelstein JM, Wold LE (2021). Double trouble: Combined cardiovascular effects of particulate matter exposure and coronavirus disease 2019. Cardiovasc. Res., 117(1): 85–95. https://doi.org/10.1093/cvr/cvaa293

Thiruvengadam M, Chung I-M, Samynathan R, Chandar SRH, Venkidasamy B, Sarkar T, Rebezov M, Gorelik O, Shariati MA, and Simal-Gandara J (2024). A comprehensive review of beetroot (Beta vulgaris L.) bioactive components in the food and pharmaceutical industries. Crit. Rev. Food Sci. Nutr., 64(3): 708–739. https://doi.org/10.1080/10408398.2022.2108367

Vicidomini C, Palumbo R, Moccia M, Roviello GN (2024). Oxidative processes and xenobiotic metabolism in plants: Mechanisms of defense and potential therapeutic implications. J. Xenobiot., 14(4): 1541–1569. https://doi.org/10.3390/jox14040084

Yang YM, Cho YE, Hwang S (2022). Crosstalk between oxidative stress and inflammatory liver injury in the pathogenesis of alcoholic liver disease. Int. J. Mol. Sci., 23(2): 774. https://doi.org/10.3390/ijms23020774