Assessment of Ovarian Inflammatory and Oxidative Stress Response to High-Salt Dietary Exposure: Protective Role of Allium Sativum Extract in Adult Female Wistar Rats

standard. Spectrophotometric data acquisition was conducted using a Vis Spectrophotometer S23A (Axiom, UK) for all colorimetric determinations. Statistical Analysis All data are expressed as mean ± standard error of the mean . Inter group comparisons were conducted using one-way analysis of variance via SPSS Statistics software (version 20.0; IBM Corp., Armonk, NY, USA). Statistical significance was established at a threshold of P < 0.05. Results In Table 1, TNF-alpha was significantly higher in the HSD-only group compared to the NC. In the groups that were given AS extract at 100, 200 and 400 mg/kg, the level of TNF-alpha was significantly decreased compared to the HSD-only group. In the 200 and 400 mg/kg treated groups, the levels of TNF-alpha were significantly lower compared to the AS 100 mg/kg treated group. In Table 1, ovarian IL-10 was significantly reduced in the HSD-only group compared to the NC. Treatment with AS extract significantly improved IL-10 in a dose-dependent manner. Table 1: Results of Allium sativum extract on ovarian TNF-alpha and ovarian IL-10 Groups TNF-α (pg/mL) IL-10 (pg/mL) NC 150.64±0.75 381.52±3.64 HSD-only 190.80±2.35a 273.34±4.25a HSD+AS (100mg/kg) 177.36±1.12ab 285.38±2.17ab HSD+AS (200mg/kg) 164.46±3.79abc 307.98±3.61abc HSD+AS (400mg/kg) 148.02±2.88abcd 312.66±3.43abcd NC = normal control, HSD = high salt diet, AS =Allium sativum, superscripts a, b, c, and d represent statistically significant difference (P<0.05) compared to NC, HSD, HSD+AS 100 mg/kg and HSD+AS 200 mg/kg respectively. MDA, SOD, GSH and CAT Table 2, shows that malondialdehyde concentration was significantly elevated in the HSD- only group compared to the normal control (NC) group. Administration of AS extract at 200 and 400 mg/kg significantly reduced MDA levels relative to the HSD-only group. Although treatment with 100 mg/kg of AS extract lowered ovarian MDA (152.42 ± 0.55), the reduction was not statistically significant. Notably, the 400 mg/kg group exhibited significantly lower MDA compared to the HSD+AS (100 mg/kg) group. tissue superoxide dismutase activity was markedly decreased in the HSD-only group (16.90 ± 1.13) compared to the NC group (29.54±1.08). Treatment with AS extract at 100, 200, and 400 mg/kg significantly increased ovarian SOD levels (17.66 ± 0.45, 22.64 ± 0.66, and 22.94 ± 0.94, respectively) compared to the HSD-only group. Furthermore, SOD activity in the 400 mg/kg group was significantly higher than in the HSD+AS (100 mg/kg) group. tissue glutathione concentration was significantly reduced in the HSD-only group (11.12 ± 0.71) compared to the NC group (28.90 ± 1.50). Treatment with AS extract at 100, 200, and 400 mg/kg significantly increased ovarian GSH levels (13.80 ± 0.98, 16.90 ± 0.40, and 19.20±0.73, respectively) relative to the HSD-only group. tissue catalase activity was significantly decreased in the HSD-only group (3.20 ± 0.10) compared to the NC group (7.90 ± 0.27). Treatment with AS extract at 400 mg/kg significantly increased ovarian CAT activity (5.84 ± 0.26) compared to the HSD-only group, as well as the HSD+AS (100 mg/kg) and HSD+AS (200 mg/kg) groups. Table 2: Results of High salt diet on oxidative stress treated with Allium sativum in adult Female Wistar rats Groups MDA (nmol/mg protein) SOD (U/mg protein) GSH (U/mg protein) CAT (U/mg protein) NC 92.60±3.50 30.66 ± 0.97 29.54 ± 1.16 7.64 ± 0.35 HSD-only 170.92±2.14a 17.26 ± 0.69a 12.46 ± 0.89a 3.44± 0.23a HSD+AS (100mg/kg) 153.28±1.50ab 18.10 ± 0.26a 14.66 ± 0.71a 3.60 ± 0.20a HSD + AS (200mg/kg) 147.78±1.41abc 22.34 ± 1.02abc 17.52 ± 0.49ab 4.90 ± 0.16abc HSD + AS (400mg/kg) 139.98±1.12abc 23.32 ± 0.59abc 19.32 ± 0.70abc 5.86 ± 0.19acd NC = normal control, HSD = high salt diet, AS =Allium sativum, superscripts a, b, c, and d represent statistically significant difference (P<0.05) compared to NC, HSD, HSD+AS 100 mg/kg and HSD+AS 200 mg/kg respectively. Discussion Female reproductive health is profoundly influenced by dietary and environmental factors, with excessive salt intake emerging as a critical disruptor of immune and oxidative balance (Li et al., 2019). The ovary and uterus, being hormonally regulated and metabolically active tissues, are particularly vulnerable to such perturbations (Barraza-Ortega et al., 2025). In this study, high-salt dietary exposure induced a pronounced pro-inflammatory state and oxidative stress. The cytokine profile observed in the HSD-only group revealed a significant rise in TNF-α, a master regulator of inflammation, alongside a reduction in IL-10, a potent anti-inflammatory cytokine. This imbalance reflects a shift toward a pro-inflammatory milieu, which can compromise function by disrupting follicular development, steroidogenesis, and uterine receptivity (Fitzgerald et al., 2020; Chen et al., 2024). Treatment with Allium sativum (AS) extract effectively attenuated TNF-α levels and restored IL-10 expression in a dose-dependent manner. These effects are likely mediated by garlic’s bioactive constituents; allicin and polyphenols which inhibit NF-κB activation, thereby suppressing pro-inflammatory cytokine transcription, while simultaneously enhancing STAT3- driven IL-10 production (Ciou et al., 2024; Chen et al., 2024). The dual modulation of TNF-α and IL-10 highlights AS extract’s capacity to recalibrate immune responses, preserving reproductive homeostasis under salt-induced stress (Mans, 2023). Oxidative stress markers in uterine tissue further substantiated the impact of HSD. Elevated MDA levels indicated enhanced lipid peroxidation, while reductions in SOD, GSH, and CAT reflected compromised antioxidant defenses (Halliwell & Gutteridge, 2015). These changes are characteristic of ROS-driven tissue injury, which can impair function and reproductive outcomes (Rahman et al., 2021). Administration of AS extract significantly mitigated these effects, lowering MDA and enhancing antioxidant enzyme activities in a dose-dependent manner. Garlic’s antioxidant efficacy is well-documented, with allicin and related sulfur compounds acting directly as free radical scavengers and indirectly by activating the Nrf2 pathway, which upregulates antioxidant gene expression (Mahajan, 2025; Chen et al., 2025). The restoration of SOD, GSH, and CAT observed in this study emphasizes the ability of AS extract to strengthen endogenous antioxidant defenses, thereby counteracting the oxidative burden imposed by high salt intake. Notably, the highest dose (400 mg/kg) produced the most pronounced improvements, suggesting a threshold for optimal bioactivity (Das et al., 2024). These findings reveal a coherent mechanistic narrative: high salt intake disrupts immune and redox balance in reproductive tissues, while Allium sativum extract restores homeostasis through dual modulation of inflammatory and antioxidant pathways. The suppression of TNF-α and enhancement of IL-10 reflect inhibition of NF-κB and activation of STAT3 signaling, respectively, while the reduction in MDA and restoration of antioxidant enzymes align with Nrf2-mediated transcriptional regulation (Arreola et al., 2025). This integrated response highlights the capacity of AS extract to simultaneously dampen inflammatory cascades and reinforce antioxidant defenses, thereby protecting tissues from diet- induced injury. References Abdelnour, S. A., Abd El-Hack, M. E., Noreldin, A. E., Batiha, G. E., Beshbishy, A. M., Ohran, H., ... & Swelum, A. A. (2020). High salt diet affects the reproductive health in animals: an overview. Animals, 10(4), 590. Agarwal, A., Gupta, S., & Sharma, R. K. (2005). Role of oxidative stress in female reproduction. Reproductive biology and endocrinology, 3(1), 28. Arreola, R., Quintero-Fabián, S., López-Roa, R. I., Flores-Gutiérrez, E. O., Reyes-Grajeda, J. P., Carrera-Quintanar, L., & Ortuño-Sahagún, D. (2025). Immunomodulation and anti‐ inflammatory effects of garlic compounds. Journal of immunology research, 2015(1), 401630. Barraza-Ortega, E., Gómez-Gil, B., García-Gasca, T., Lizárraga, D., Díaz, N., & García-Gasca, A. (2025). The impact of lifestyle on reproductive health: microbial complexity, hormonal dysfunction, and pregnancy outcomes. International Journal of Molecular Sciences, 26(17), 8574. Behrman, H. R., Kodaman, P. H., Preston, S. L., & Gao, S. (2001). Oxidative stress and the ovary. Journal of the Society for Gynecologic Investigation, S40-S42. Chen, H., Song, J., Zeng, L., Zha, J., Zhu, J., Chen, A., ... & Chen, G. (2024). Dietary sodium modulates mTORC1-dependent trained immunity in macrophages to accelerate CKD development. Biochemical pharmacology, 116505. Chen, L., Peng, X. L., Chen, Z. X., Qi, L. M., Deng, T. T., & Xia, L. N. (2024). Immune dysregulation orchestrated by high-salt diet: Mechanistic insights into disease pathogenesis. Nutrition and Dietary Supplements, 147-164. Chen, M., Li, Y., & Liu, X. (2025). A review of the role of bioactive components in legumes in the prevention and treatment of cardiovascular diseases. Food & Function, 16(3), 797-814. Ciou, J. J., Chien, M. W., Hsu, C. Y., Liu, Y. W., Dong, J. L., Tsai, S. Y., ... & Sytwu, H. K. (2024). Excess Salt Intake Activates IL-21–Dominant Autoimmune Diabetogenesis via a Salt-Regulated Ste20-Related Proline/Alanine-Rich Kinase in CD4 T Cells. Diabetes, 73(4), 592-603. Craig, W. (1999). Health-promoting properties of common herbs. The American journal of clinical nutrition, 70(3), 491S-9S. Das, D., M, K., Mitra, A., Zaky, M. Y., Pathak, S., & Banerjee, A. (2024). A review on the efficacy of plant-derived bio-active compounds curcumin and aged garlic extract in modulating cancer and age-related diseases. Current Reviews in Clinical and Experimental Pharmacology Formerly Current Clinical Pharmacology, 146-162. Fitzgerald, T., Gupta, A., & Sampson, M. . (2020). Impact of high-sodium diets on the hypothalamic-pituitary-gonadal axis in female rats: A mechanistic review. Endocrine Journal, 67(6), 487-497. Halliwell, B., & Gutteridge, J. M. (2015). Free radicals in biology and medicine. Oxford university press. Hou, E., Yan, J., Zhu, X., & Qiao, J. (2022). High-salt diet contributes to excess oxidative stress and abnormal metabolism in mouse ovaries. Biomedical Chromatography, 36(12), e5500. Hu, Y., Yang, X., & Lin, Z. (2021). High sodium intake impairs function and induces oxidative stress in female rats. Journal of Endocrinology, 248(3), 261–272. Khatua, T. N., Adela, R., & Banerjee, S. K. (2013). Garlic and cardioprotection: insights into the molecular mechanisms. Canadian journal of physiology and pharmacology, 91(6), 448- 458. Li, M., Wang, J., & Liu, Q. (2019). High-salt diet, renal function, and reproductive health: A review of recent findings. Reproductive Toxicology, 88, 23–32. Mahajan, B. (2025). Molecular Mechanisms and Therapeutic Implications of Bioactive Substances from Medicinal Plants in Type 2 Diabetes. Sciences, 131-138. Mans, D. R. (2023). Naturally Occurring Antioxidants in Seven Well-Known Fruits from the Republic of Suriname (South America): Part 2. In Recent Developments in Antioxidants from Natural Sources. IntechOpen. O'Hara, M., Kiefer, D., Farrell, K. & Kemper, K. (1998). A review of 12 commonly used medicinal herbs. Archives of family medicine, 7(6), 523. Perić, L., Drenjančević, I., Jukić, I., Boris, A., Šušnjara, P., Kolobarić, N., ... & Stupin, A. (2025). Carnosine-enriched functional food enhances micro-and macrovascular endothelium- independent vasodilation in competitive athletes—a randomized study. Applied Physiology, Nutrition, and Metabolism, 50, 1-11. Rahman, M. A., Hannan, M. A., Dash, R., Rahman, M. H., Islam, R., Uddin, M. J., ... & Rhim, H. (2021). Phytochemicals as a complement to cancer chemotherapy: Pharmacological modulation of the autophagy-apoptosis pathway. Frontiers in Pharmacology, 639. Ruder, E. H., Hartman, T. J., & Goldman, M. B. (2009). Impact of oxidative stress on female fertility. Current opinion in obstetrics and gynecology, 21(3), 219-222. Vassalle, C., Maffei, S., & Fiorentino, G. . (2015). Impact of dietary salt on oxidative stress and cardiovascular health: Implications for female reproductive organs. Oxidative Medicine and Cellular Longevity, 295871. Yang, L., Zuo, X., & Zheng, L. . (2018). Alterations in morphology and function due to excessive salt consumption in rats. Reproductive Toxicology, 113–121.