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. 2025 Jun 16;35(1):22.
doi: 10.1186/s12610-025-00271-4.

The antioxidant and therapeutic effects of Malva sylvestris extract on testicular tissue and sperm quality in varicocele-induced adult Wistar rats

Fatemeh Koohkan  1 , Mahnaz Azarnia  1 , Latifeh Karimzadeh Bardeei  2
Affiliations

The antioxidant and therapeutic effects of Malva sylvestris extract on testicular tissue and sperm quality in varicocele-induced adult Wistar rats

Fatemeh Koohkan et al. Basic Clin Androl. .

Abstract
in English, French

Background: Varicocele is a major male infertility issue, and Malva sylvestris shows promise as a treatment due to its antioxidant properties. The present study evaluated the protective effects of Malva sylvestris on testicular health, sperm quality, and oxidative stress-related gene expression. Adult male Wistar rats were randomly assigned into five groups (n = 8): the control group, the varicocele model group, the varicocele group with partial occlusion of the left renal vein treated with 750 or 1500 mg/kg Malva sylvestris for 21 days, and a surgical sham group. The epididymal content and Histological analyses of animals testicular tissue were examined to evaluate fertility parameters, and qRT-PCR was employed to determine the expression of SIRT1, FOXO1, NRF2, NF-κB, and TGF-β genes.

Results: Varicocele leads to the induction of apoptosis, the occurrence of DNA damage, a reduction in SIRT1 and FOXO1, and an increase in NRF2, TGF-β, and NF-κB gene expression. According to Histological morphometric analysis, treatment with Malva sylvestris showed the increased thickness of Mean Seminiferous Tubule Diameter and Epithelial Thickness in the spermatogenic epithelium, as well as the presence of a greater number of germ cells and mature sperm. In this study, Malva sylvestris treatment showed antioxidative effects since it upregulated the expression of SIRT1 and FOXO1 genes and downregulated the expression of NF-κB, NRF2, and TGF-β genes.

Conclusion: Based on the findings, Malva sylvestris as cytoprotective agent modulate key antioxidant pathways, including upregulation of SIRT1 and FOXO1 (linked to cellular repair and OS resistance) and downregulation of pro-inflammatory mediators; then, Malva sylvestris as a promising natural antioxidant for managing varicocele-related infertility, offering a potential adjunct or alternative to conventional therapies.

RéSUMé: CONTEXTE: La varicocèle est un problème majeur d’infertilité masculine, et Malva sylvestris est prometteuse en tant que traitement, en raison de ses propriétés antioxydantes. La présente étude a évalué les effets protecteurs de Malva sylvestris sur la santé testiculaire, la qualité du sperme et l’expression des gènes liés au stress oxydatif. Des rats Wistar mâles adultes ont été répartis au hasard en cinq groupes (n = 8) : groupe témoin, groupe modèle varicocèle, groupe varicocèle avec occlusion partielle de la veine rénale gauche traité avec 750 ou 1500 mg/kg de Malva sylvestris pendant 21 jours, et groupe simulé chirurgicalement. Le contenu épididymaire et les analyses histologiques de tissu testiculaire des animaux ont été examinés pour évaluer les paramètres de fertilité, et la qRT-PCR a été utilisée pour déterminer l’expression des gènes SIRT1, FOXO1, NRF2, NF-κB et TGF-β. RéSULTATS: La varicocèle entraîne l’induction d’une apoptose, l’apparition de dommages à l’ADN, une réduction de SIRT1 et FOXO1, et une augmentation de l’expression des gènes NRF2, TGF-β et NF-κB. Selon l’analyse morphométrique histologique, le traitement par Malva sylvestris a montré une augmentation de la moyenne de l’épaisseur du tube séminifère et de l’épaisseur épithéliale de l’épithélium séminifère, ainsi que la présence d’un plus grand nombre de cellules germinales et de spermatozoïdes matures. Le traitement par Malva sylvestris a montré des effets antioxydants puisqu’il régulait à la hausse l’expression des gènes SIRT1 et FOXO1 et à la baisse l’expression des gènes NF-κB, NRF2 et TGF-β. CONCLUSIONS: Sur la base des résultats, Malva sylvestris, en tant qu’agent cytoprotecteur, module les principales voies antioxydantes, y compris la régulation à la hausse de SIRT1 et FOXO1, et la régulation à la baisse des médiateurs pro-inflammatoires. Malva sylvestris, en tant qu’antioxydant naturel prometteur pour la gestion de l’infertilité liée à la varicocèle, offre un complément potentiel ou une alternative aux thérapies conventionnelles.

Keywords: Malva sylvestris; Infertility; Oxidative stress; Sperm; Testis; Varicocele.

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Conflict of interest statement

Declarations. Ethics approval and consent to participate: There were no “human subjects” in this study. All animal experiments comply with the ARRIVE guidelines and carried out in accordance with the National Institutes of Health guide for the care and use of Laboratory animals. The study was approved by the Ethics Committee of Kharazmi University under code IR.KHU.REC.1402.091. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
A Surgery and induction of the varicocele model in adult male Wistar rats. The left renal vein, from which the internal spermatic vein branches off, was tied. a: Location of the renal vein and internal spermatic vein relative to the kidney, b: needle passage to isolate the renal vein, c: renal vein node. 1: Renal vein, 2: spermatic vein. B Histological analysis of the testicular tissue of the studied groups using hematoxylin and eosin staining. a: Control group, b: varicocele group, c: M.S 750 mg/kg treatment group, d: M.S 1500 mg/kg treatment group, 1: Spermatogonial cell, 2: primary spermatocyte cell, 3: secondary spermatocyte cell, 4: spermatid, 5: sperm. C Histological analysis of a: mean seminiferous tubule diameter, b: mean seminiferous epithelial thickness. D The number of spermatogonia, primary spermatocytes, and spermatids. The extent of destruction in the induction group and repair in the treatment group can be seen. Differences among the experimental groups were analyzed using one-way analysis of variance (ANOVA) coupled with Tukey’s post hoc test. All values are expressed as mean ± SD. A p value of ≤ 0.05 was considered significant. P < 0.05(*), P < 0.01(**), P < 0.001(***). M.S: Malva sylvestris
Fig. 2
Fig. 2
Comparison of epididymal sperm parameters. a: Motility, b: viability, c: normal sperm morphology in the left testicle. The treatment groups showed a significant increase compared to the induction group. M.S 750: M.S 750 mg/kg, M.S 1500: M.S 1500 mg/kg, ns = 8. Differences among the experimental groups were analyzed using one-way analysis of variance (ANOVA) coupled with Tukey’s post hoc test. A p value of ≤ 0.05 was considered significant. P < 0.05(*), P < 0.01(**), P < 0.001(***). M.S: Malva sylvestris
Fig. 3
Fig. 3
Effects of M.S treatment (750 and 1500 mg/kg) on Sperm Chromatin Structure in varicocele-induced rats. A Comparison of sperm DFI strand damage through sperm chromatin structure analysis. B Comparison of DFI in the studied groups. C Comparison of the level of apoptosis by examining the MMP of the tissue testis with JC-1 dye in a: the control group, b: the varicocele group, c: the 750 mg/kg treatment group, and d: the 1500 mg/kg group. The extent of damage in the induction group and repair in the treatment group can be seen in this image, according to the chromatin structure (DFI) and mitochondrial membrane staining. Differences among the experimental groups were analyzed using one-way analysis of variance (ANOVA) coupled with Tukey’s post hoc test. A p value of ≤ 0.05 was considered significant. P < 0.05(*), P < 0.01(**), P < 0.001(***). M.S: Malva sylvestris; DFI: DNA Fragmentation Index; SCSA: Sperm Chromatin Structure Assay; DFI: DNA Fragmentation Index
Fig. 4
Fig. 4
Comparison of sperm cell apoptosis in the investigated groups. a Control group, b varicocele group, c 750 mg/kg treatment group, d 1500 mg/kg treatment group, e comparison of sperm cell necrosis in the investigated groups, f comparison of secondary apoptosis in sperm cells in the studied groups, g comparison of living sperm cells in the studied groups, h comparison of primary apoptosis in sperm cells in the studied groups. Differences in live and apoptotic cells are observed in the induction and treatment groups using one-way analysis of variance (ANOVA) coupled with Tukey’s post hoc test. A p value of ≤ 0.05 was considered significant. P < 0.05(*), P < 0.01(**), P < 0.001(***). M.S 750: M.S 750 mg/kg, M.S 1500: M.S 1500 mg/kg. Q1: Necrosis, Q2: Late apoptosis, Q3: Early apoptosis, Q4: Live cells. M.S: Malva sylvestris
Fig. 5
Fig. 5
Effects of M.S treatment (750 and 1500 mg/kg) on SIRT1, FOXO1, NF-κB, and TGF-β gene expression in the varicocele model. a SIRT1 gene expression, b NRF2 gene expression, c NF-κB gene expression, d TGF-β gene expression, e FOXO1 gene expression. Malva Sylvestris shows increased expression of SIRT1 and FOX01 genes and decreased expression of NF-kB, TGF-β, and NRF2 genes. Differences among the experimental groups were analyzed using one-way analysis of variance (ANOVA) coupled with Tukey’s post hoc test. A p value of ≤ 0.05 was considered significant. P < 0.05(*), P < 0.01(**), P < 0.001(***). M.S: Malva sylvestris
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References

    1. Babakhanzadeh E, Nazari M, Ghasemifar S, Khodadadian A. Some of the factors involved in male infertility: a prospective review. Int J Gen Med. 2020;13:29–41. Available from: https://www.tandfonline.com/action/journalInformation?journalCode=dijg20. Cited 2023 Jun 7. - PMC - PubMed
    1. Alharbi M, Zini A. Epidemiology of varicocele in pediatric, adolescent, and adult populations. Varicoc Male Infertil. 2019;70:97–106. Available from: https://link.springer.com/chapter/10.1007/978-3-319-79102-9_8. Cited 2023 Jun 9. - DOI
    1. Moein MR, Soleimani M, Tabibnejad N. Reactive oxygen species (ROS) production in seminal fluid correlate with the severity of vericocele in infertile men. Iranian J Reprod Med. 2008;6:15.
    1. Rao L, Babu A, Kanakavalli M, Padmalatha V, Singh A, Singh PK, et al. Chromosomal abnormalities and Y chromosome microdeletions in infertile men with varicocele and idiopathic infertility of South Indian Origin. J Androl. 2004;25:147–53. - PubMed
    1. Wang K, Gao Y, Wang C, Liang M, Liao Y, Hu K. Role of oxidative stress in varicocele. Front Genet. 2022;13:850114. - PMC - PubMed

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