Abstract
Infertility rates have risen significantly, one of which is due to monosodium glutamate (MSG) consumption. Recent studies have shown that flavonoids in black garlic (Allium sativum) act as antioxidants. The aim of this study was to assess the effect of black garlic extract (BGE) on gonadosomatic index, follicle-stimulating hormone (FSH) levels, and spermatozoa quality in rats exposed to MSG. Twenty-five healthy rats, aged ten to twelve weeks, were divided equally into five experimental groups: (1) negative control (NC), no intervention; (2) positive control (PC), fed with MSG 8 mg/kg; and (3) fed with MSG + BGE 200 mg/kg; (4) fed with MSG + BGE 400 mg/kg; and (5) fed with MSG + BGE 600 mg/kg. Oral MSG was administered once a day for two weeks before BGE administration was started for two weeks. The measured endpoints were gonadosomatic index, FSH levels, and spermatozoa concentration and quality (spermatozoa motility and abnormality). Analysis of variance (ANOVA) followed by Duncan’s post hoc analysis was used to assess the measurement differences. The result suggested that the administration of BGE did not significantly affect the gonadosomatic index (p=0.513). Significant decreases in FSH levels (p=0.005) and spermatozoa concentration were observed in the PC group compared to other groups (p<0.001). Additionally, spermatozoa motility was significantly lower in the PC group compared to NC, BGE200, BGE400, and BGE600 (p<0.001), with higher motility noted in BGE200, BGE400, and BGE600 compared to PC (p<0.001). Furthermore, PC had significantly higher spermatozoa abnormalities compared to NC, BGE200, BGE400, and BGE600 (p<0.001). In conclusion, administration of BGE had a significant effect on the improvement of FSH levels and the quality of spermatozoa in rats exposed to MSG.
Keywords: Black garlic, MSG, gonadosomatic index, FSH, spermatozoa quality
Introduction
Infertility rates increased annually by 12–15% globally. In Indonesia, 10–15% of 39.8 million childbearing-age couples faced infertility issues [1,2]. Oxidative stress contributes to infertility, with approximately 30–80% originating from the environment and dietary lifestyles involving monosodium glutamate (MSG) consumption [3,4]. Developed countries consume MSG at 0.3– 1.0 g/day, while developing countries, such as Indonesia, have an intake of 0.6 g/day [5,6].
Glutamate in MSG has a direct free radical effect at a cellular level, causing damage to testicular organs and spermatozoa cells [7]. MSG-induced oxidative stress leads to deoxyribonucleic acid (DNA) damage, cell membrane peroxidation, and abnormal spermatozoa morphology, eventually leading to cell death [8,9]. In addition, excessive MSG-induced lipid peroxidation damages mitochondrial DNA, reducing spermatozoa motility and causing infertility by hindering penetration of sperm cells into ovum cells [10,11].
Free radicals from MSG can also affect pituitary cells, disrupting reproductive and endocrine systems [12]. Pituitary cell damage inhibits the production of gonadotropin-releasing hormone (GnRH), leading to decreased luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels [12]. Reduced FSH levels hinder spermatogenesis, leading to decreased spermatozoa numbers [13]. Furthermore, diminished LH and FSH impact Leydig and Sertoli cell activity, causing atrophy in testicles and a decline in testicular weight and gonadosomatic index over time [12,14].
Recent studies have shown that antioxidants effectively inhibit oxidative stress, promoting and sustaining male fertility [15-17]. Extensive studies have been conducted on herbal plants as a source of antioxidants, including the utilization of black garlic (Allium sativum) [15-18]. Black garlic, produced by heating garlic at 70°C for 15, 25, and 35 days, has higher antioxidant content than raw garlic due to the presence of phenolic compounds of flavonoids and S-Allylcysteine [16]. The aim of this study was to assess the effect of black garlic extract (BGE) on gonadosomatic index, FSH level, and spermatozoa quality in rats exposed to MSG.
Methods
Study design
An in vivo experimental study was conducted from August 2022 to September 2023 at the Laboratory of Experimental Animal and Physiology, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia. Federer equation was employed for sample size calculation, resulting in 25 rats for five experimental groups–divided equally to each group. Randomization was carried out using a completely randomized design method with a unidirectional pattern. Each experimental unit was homogeneous and each unit had the same opportunity for each intervention.
Eligibility criteria
Wistar rats (Rattus norvegicus) were obtained from the Laboratory of Experimental Animal and Physiology, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia. Healthy rats, aged ten to twelve weeks and weighing 150–200 grams, were included. Exclusion criteria were rats with ongoing infection and the presence of structural or functional abnormalities. Drop-out criteria included the presence of any complication, infection, and death during the study period.
Animal preparations
Rats were caged for seven days in a consistent environment, well-shaded, and tranquil rooms. The animals were provided with the same quantity and type of food, with a tray beneath the cages to collect urine and feces, cleaned once daily. Each rat was accommodated in ventilated cages maintained at a temperature of 22°C and humidity ranging from 50–60%. A regular diurnal lighting cycle (12:12 light-dark) was maintained, with lights turning on at 5:30 AM. Rats were fed commercial ratio brand 552 at 10% of the rat’s average body weight, approximately 15–20 grams per day. Rats were given uncontaminated clean water ad libitum.
Study groups and interventions
Rats were randomly grouped into five groups: (1) negative control (NC), no intervention; (2) positive control (PC), provided with MSG 8 mg/kg only; (3) BGE200, provided with MSG 8 mg/kg and BGE 200 mg/kg; (4) BGE400, MSG 8 mg/kg and BGE 400 mg/kg; and (5) BGE600, MSG 8 mg/kg and BGE 600 mg/kg. MSG was administered for two weeks, of which it was dissolved in 10 mL of 0.5% carboxymethyl cellulose (CMC) and administered once a day orally. BGE200, BGE400, and BGE600 groups received BGE with specific doses once a day orally for two weeks. All animals were then cervical decapitated.
Outcome endpoints
The endpoints of the study were the gonadosomatic index, FSH level, and spermatozoa quantity and quality (spermatozoa motility and abnormality). The gonadosomatic index was calculated by weighing both testicular organs, dividing by the rat’s body weight, and multiplying by 100%. Serum FSH level was measured using enzyme-linked immunosorbent assay (ELISA) using Rat Follicle-Stimulating Hormone Competitive Elisa Kit (BZ-08185100-CPEB, Biozatix Indonesia, Indonesia). The blood was collected using a cardiac puncture. The absorbance was read on a spectrophotometer at a wavelength of 450 nm [19].
Spermatozoa were obtained from epididymal secretions in 0.9% NaCl to assess the spermatozoa quality. Spermatozoa concentration and motility were assessed using wet preparations in a Neubauer counting chamber (400× magnification) and recorded with an OptiLab camera on a microscope. The concentration value of spermatozoa was expressed in mm3. Evaluation of spermatozoa motility was carried out by observing sperm movement directly under a microscope and grouped into moving groups (both fast and slow moving) and groups that did not move at all. Spermatozoa morphology (spermatozoa abnormality) was analyzed from photographs of smears stained with eosin 2%. The percentages of normal and abnormal spermatozoa were counted from every 200 spermatozoa observed. Spermatozoa cells with a head without an acrosome, a neck that bends and/or folds, and a tail that bends and/or twists were considered abnormal as recommended [20,21].
Statistical analysis
Shapiro-Wilk test was employed for assessing the normality of data distribution. To assess differences in gonadosomatic index, FSH level, spermatozoa concentration and quality (motility and abnormality), the analysis of variance (ANOVA) test followed by Duncan’s post hoc test was used. Statistical analysis was conducted using SPSS version 25.0 software (IBM SPSS, Chicago, IL, USA), with p<0.05 was considered statistically significant.
Results
Effect of black garlic on gonadosomatic index
Our data suggested that the administration of BGE did not significantly affect the gonadosomatic index of rats (p=0.513) (Table 1). The MSG-treated group (positive control) had the smallest gonadosomatic index, while the negative control group had the highest. The negative control had 15.2%, 2.2%, 8.4%, and 10.1% higher gonadosomatic indexes compared to the positive control, BGE200, BGE400, and BGE600 groups, respectively.
Table 1.
Effects of black garlic extract on gonadosomatic index of rats (n=25)
| Group | Gonadosomatic index (%) (±SD) | p-value |
|---|---|---|
| Negative control | 1.78±0.24 | 0.513 |
| Positive control | 1.51±0.19 | |
| BGE200 | 1.74±0.33 | |
| BGE400 | 1.63±0.30 | |
| BGE600 | 1.60±0.21 |
Effect of black garlic on FSH level
Our data indicated that the MSG-treated only group (positive control) had lower FSH levels compared to the non-treated group (Table 2). Administration of BGE could increase FSH levels in rats exposed to MSG (Table 2). There was a significant difference in FSH levels among groups (p=0.005). The FSH level in the negative control group was 36.6% lower compared to the positive control group, while the FSH concentration in groups BGE200, BGE400, and BGE600 was higher by 5.7%, 29.3%, and 1.63% compared to the positive control group, respectively. Post-hoc sanalysis indicated that the positive control group had lower FSH levels compared to other groups significantly (Figure 1).
Table 2.
Effect of black garlic on follicle-stimulating hormone (FSH) level of rats (n=25)
| Group | FSH level (ng/mL) (±SD) | p-value |
|---|---|---|
| Negative control | 1.23±0.22 | 0.005 |
| Positive control | 0.78±0.34 | |
| BGE200 | 1.30±0.17 | |
| BGE400 | 1.59±0.42 | |
| BGE600 | 1.25±0.38 |
Figure 1.
Post-hoc analysis showing the comparison of follicle-stimulating hormone (FSH) levels between groups of animals. Different superscripts indicate significant differences (p<0.05). BGE: black garlic extract; NC: negative control; PC: positive control.
Effect of black garlic on spermatozoa quantity and quality
Our data suggested that administration of BGE could increase spermatozoa concentration in rats exposed to MSG (Table 3). The positive control group had the lowest spermatozoa concentration, while the BGE600 group had the highest concentration. There was a significant difference in rat spermatozoa concentration among groups (p<0.001) (Table 3). Post-hoc analysis indicated that spermatozoa concentration in the positive control group was significantly 72.27% lower compared to negative control (p<0.001). The level of spermatozoa in groups BGE200, BGE400, and BGE600 was lower at 40.23%, 13.51%, and 2.73% compared to the negative control, BGE200, and BGE400 groups that were significantly lower than the negative control, respectively (Figure 2A).
Table 3.
Effect of black garlic on spermatozoa quantity and quality of rats (n=25)
| Parameters | Negative control | Positive control | BGE200 | BGE400 | BGE600 | p-value |
|---|---|---|---|---|---|---|
| Spermatozoa concentration (106/mm3) | 139.20±10.06 | 38.60±6.80 | 83.20±15.67 | 120.40±14.94 | 135.40±4.51 | <0.001 |
| Spermatozoa motility (%) | 70.00±10.00 | 46.00±11.40 | 75.00±5.00 | 59.00±10.24 | 47.00±13.03 | <0.001 |
| Spermatozoa abnormality (%) | 31.76±21.07 | 70.07±3.86 | 34.96±1.44 | 39.34±14.03 | 57.70±5.56 | <0.001 |
BGE: black garlic extract
Figure 2.
Post-hoc analysis showing the comparison of spermatozoa concentration (A), spermatozoa motility (B), and spermatozoa morphology (C) between groups of animals. Groups with different superscripts indicate significant differences (p<0.05). BGE: black garlic extract; NC: negative control; PC: positive control.
Our data also suggested that MSG reduced spermatozoa motility, while BGE could increase spermatozoa motility in rats exposed to MSG (Table 3). There was a significant difference in rat spermatozoa motility among groups (p<0.001). The motility of spermatozoa in the positive control, BGE400, and BGE600 groups was 34.28%, 15.71%, and 32.85% lower, respectively, compared to the negative control group, while the motility of spermatozoa in BGE200 group was 7.14% was higher than the negative control group. Post-hoc analysis indicated that the motility of rat sperm between PC and BGE200 groups had significant differences (p<0.001). The motility of spermatozoa between PC, BGE400, and BGE600 showed no significant difference (Figure 2B).
In addition, our data suggested that MSG increased spermatozoa abnormalities, while BGE reduced spermatozoa abnormalities in rats exposed to MSG (Table 3). The BGE200 group had the lowest spermatozoa abnormalities, while the positive control group had the highest (Table 3). The morphology of spermatozoa abnormalities in the positive control, BGE400, and BGE600 groups were respectively 120.62%, 23.86%, and 81.57% higher compared to the negative control group, while the morphology of spermatozoa abnormalities in the BGE200 group was 10.07% higher than the negative control group. The results of statistical analysis showed that the morphology of mouse spermatozoa abnormalities between treatments showed very significant differences (p<0.001) (Table 3). Post-hoc analyses indicated that the positive control group had significantly higher spermatozoa abnormalities (p<0.001) compared to the negative control, BGE200, BGE400, and BGE600 (Figure 2C).
Discussion
The present study found that the administration of black garlic extract did not significantly affect the gonadosomatic index of rats (p>0.05). This lack of significance in the gonadosomatic index may be attributed to the relatively brief duration of MSG administration, in which the increase in testicular weight in this study was accompanied by an increase in body weight in other groups, resulting in nonsignificant differences in the gonadosomatic index. The gonadosomatic index is influenced by testicular weight and body weight of rat, with testicular weight dependent on spermatogenic cell differentiation [14].
Black garlic has main bioactive components of phenolic organic compounds, especially flavonoids, contributing as antioxidants to minimize damage to Sertoli cells in testicles [14]. However, studies on black garlic and its impact on testicular weight and gonadosomatic index are limited. A study found that oral administration of 5% raw and 15% cooked garlic for 30 days resulted in an increase in Sertoli cell count and testicular weight [18]. In addition, flavonoids in neem seed extract increased testicular weight and gonadosomatic index [22]. In contrast, another study reported a decrease in testicular weight and gonadosomatic index in rats orally administered 0.3 and 0.6 mg/gr BW of MSG for 35 days [23]. Leydig and Sertoli cells play a crucial role in spermatogenesis, and a decrease in testicular weight due to MSG toxicity in previous study may result from a reduction in germ cell and mature spermatid density, along with Leydig and Sertoli cell damage.
In the present study, administration of black garlic extract can increase FSH concentrations in rats exposed to MSG. The decrease in FSH levels in MSG-treated control in the present study is attributed to direct damage to Sertoli cells by MSG, leading to oxidative stress and membrane damage, thereby interfering with FSH receptors (FSHR) and inhibiting FSH input into Sertoli cells [24]. This finding aligned with a previous study that showed decreased FSH concentrations in rats administered 30 and 60 mg/kg BW of MSG for eight weeks compared to the control group [9]. A previous study also observed decreased testosterone concentrations in rats administered 8 mg/kg BW/day of MSG for 14 days compared to the control group [25].
Flavonoids in black garlic reduce oxidative stress by inhibiting lipid peroxidation and preventing the conversion of androgens into estrogen, ultimately increasing testosterone levels [14,26]. In the present study, an increase in FSH levels was observed in the BGE200 and BGE400 groups, indicating that flavonoid antioxidants can stimulate FSHR. This finding was aligned with a previous study that reported consuming flavonoid in Icariin dietary supplement at 100 and 200 mg/kg BW for 35 days increases testosterone and FSHR levels, and protects the Sertoli cells and blood-testis barrier [27]. A study found garlic administration at 6 mL/kg BW/day for 65 days could increase androgen receptors [28]. This is consistent with another study that found elevated androgen hormones after administration of garlic powder at 200 mg/kg BW for 42 days [29]. In contrast, a study reported a decrease in androgen levels after administration of garlic extract at 400 mg/kg BW for one month [30]. This finding may be attributed to variation in dosage or administration duration, yet research on the effect of black garlic in androgen levels, especially FSH levels, remains limited.
In the present study, administration of black garlic extract can increase spermatozoa concentration in rats exposed to MSG. Flavonoids in black garlic stabilize radicals, combat superoxide, and inhibit lipid peroxidase, thus enhancing spermatozoa quality [14,26,34]. A previous study demonstrated that the administration of garlic powder 200 mg/kg BW for 42 days could increase spermatozoa concentration and motility, and decrease morphological abnormalities [35]. Consistent findings were observed in another study, which emphasized the positive impact of garlic on spermatozoa concentration, motility, and viability [18,36].
On the other hand, the MSG-treated only group in the present study has the lowest spermatozoa concentration, which might be due to the oxidative stress of MSG. This finding was aligned with a previous study that showed decreased motility yet normal morphology in rats exposed to 4 mg/gr BW of MSG for 35 days [33]. Small cytoplasm content in sperm cells leads to insufficient antioxidant defenses during oxidative stress. Excessive lipid peroxidase formation damages mitochondrial DNA, contributing to decreased spermatozoa motility. This process causes a loss of 60% of polyunsaturated fatty acids, disrupting sperm membrane permeability and depleting sperm cell ATP, affecting flagellum movement. Oxidative DNA damage and peroxidation also increase abnormal morphology and death in sperm cells, ultimately reducing spermatozoa concentration [31,32].
MSG can cause damage to the hypothalamus, leading to pituitary cell damage, inhibition of GnRH production, and a decrease in the production of FSH levels [12,13]. The resulting decrease in FSH levels can affect spermatogenesis and lead to a reduction in spermatozoa concentration [12,13]. Furthermore, a decrease in FSH levels leads to a decline in Leydig and Sertoli cell activity, causing testicular atrophy, decreased testicular weight, and a decline in gonadosomatic index percentage [12,13]. Hence, long, continuous exposure to MSG can be attributed to testicular atrophy and a decrease in gonadosomatic index percentage [12,13].
Conclusion
Administration of BGE had a significant effect on the improvement of FSH levels and quality of spermatozoa in rats exposed to MSG. However, BGE treatment did not yield a significant effect on the gonadosomatic index.
Acknowledgments
Thanks to the Laboratory of Experimental Animal and Physiology, Faculty of Veterinary Medicine, Universitas Syiah Kuala and National Research Center (BRIN) for the assistance in this study.
Ethics approval
This study protocol was approved by the Committee of Research Ethics, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia (Approval number: 175/KEPH/X/2022).
Competing interests
All the authors declare that there are no conflicts of interest.
Funding
This study received no external funding.
Underlying data
All data underlying the results is available as part of the article and no additional source data is required.
How to cite
Bani II, Zulkarnain Z, Gholib G, et al. Effect of black garlic (Allium sativum) on gonadosomatic index, follicle-stimulating hormone level and spermatozoa quality: A study in monosodium glutamate-exposed rat models. Narra J 2024; 4 (2): e617 - http://doi.org/10.52225/ narra.v4i2.617.
References
- 1.Mascarenhas MN, Flaxman SR, Boerma T, et al. National, regional, and global trends in infertility prevalence since 1990: A systematic analysis of 277 health surveys. PLoS Med 2012;9(12):1–12. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2.Bennett LR, Wiweko B, Bell L, et al. Reproductive knowledge and patient education needs among Indonesian women infertility patients attending three fertility clinics. Patient Educ Couns 2015;98(3):364–369. [DOI] [PubMed] [Google Scholar]
- 3.Agarwal A, Qiu E, Sharma R.. Laboratory assessment of oxidative stress in semen. Arab J Urol 2018;16(1):77–86. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Mondal M, Sarkar K, Nath PP, et al. Monosodium glutamate suppresses the female reproductive function by impairing the functions of ovary and uterus in rat. Environ Toxicol 2018;33(2):198–208. [DOI] [PubMed] [Google Scholar]
- 5.Hajihasani MM, Soheili V, Zirak MR, et al. Natural products as safeguards against monosodium glutamate-induced toxicity. Iran J Basic Med Sci 2020;23(4):416–430. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6.Sulastri S. Analysis of MSG levels in instant noodle seasoning. Implement Sci 2014;3(1):1–24. [Google Scholar]
- 7.Kalsum U, Ilyas S, Hutahaean S.. Effect of vitamin C administration on histological picture of testicles of mice (Mus musculus L.) exposed to Monosodium glutamate (MSG). Saintia Biologi 2013;1(3):7–12. [Google Scholar]
- 8.Sukmaningsih AASGA, Ermayanti IGAM, Wiratmini NI, et al. Disruption of spermatogenesis after administration of monosodium glutamate in mice (Mus musculus L). J Biol 2011:15(2):49–52. [Google Scholar]
- 9.Kianifard D, Saiah GV, Rezaee F.. Study of the protective effects of quince (Cydonia oblonga) leaf extract on fertility alterations and gonadal dysfunction induced by monosodium glutamate in adult male Wistar rats. Rom J Diabetes Nutr Metab Dis 2015;22(4):375–384. [Google Scholar]
- 10.Tobaben S, Grohm J, Seiler A, et al. Bid-mediated mitochondrial damage is a key mechanism in glutamate-induced oxidative stress and AIF-dependent cell death in immortalized HT-22 hippocampal neurons. Cell Death Differ 2011:18(2):282–292. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11.Niederberger C. Male infertility. J Urol 2010;184(5):2085–2088. [DOI] [PubMed] [Google Scholar]
- 12.Oluwole DT, Ebiwonjumi OS, Ajayi LO, et al. Disruptive consequences of monosodium glutamate on male reproductive function: A review. Curr Res Toxicol 2024;6:100148. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13.Edward Z. Effect of monosodium glutamate (MSG) administration in male rats (Rattus norvegicus) on FSH and LH. Maj Kedokt Andalas 2010;34(2):160–166. [Google Scholar]
- 14.Ahmed T, Wang CK. Black garlic and its bioactive compounds on human health diseases: A review. Molecules 2021:26(16):1–38. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Ménézo YJ, Hazout A, Panteix G, et al. Antioxidants to reduce sperm DNA fragmentation: An unexpected adverse effect. Reprod Biomed Online 2007;14(4):418–421. [DOI] [PubMed] [Google Scholar]
- 16.Qiu Z, Zheng Z, Zhang B, et al. Formation, nutritional value, and enhancement of characteristic components in black garlic: A review for maximizing the goodness to humans. Compr Rev Food Sci Food Saf 2020;19(2):801–834. [DOI] [PubMed] [Google Scholar]
- 17.Asadpour R, Azari M, Hejazi M, et al. Protective effects of garlic aquous extract (Allium sativum), vitamin E, and N-acetylcysteine on reproductive quality of male rats exposed to lead. Vet Res Forum 2013;4(4):251–257. [PMC free article] [PubMed] [Google Scholar]
- 18.Bahrami KH, Mahjor AA, Johary H, et al. Comparative study on histopatological and histomorphometric effect of raw and cooked garlic on spermatogenesis in testis and epidydims of rats. J Fasa Univ Med Sci 2014;3:371–379. [Google Scholar]
- 19.Rose MP, Gaines Das RE, Balen AH. Definition and measurement of follicle stimulating hormone. Endocr Rev 2000;21(1):5–22 [DOI] [PubMed] [Google Scholar]
- 20.World Health Organization. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction, 4th ed. Cambridge: Cambridge University Press; 1999. [Google Scholar]
- 21.World Health Organization. WHO laboratory manual for the examination and processing of human semen, 5th ed. Geneva: World Health Organization; 2010. [Google Scholar]
- 22.Auta T, Hassan AT. Reproductive toxicity of aqueous wood-ash extract of Azadirachta indica (neem) on male albino mice. Asian Pacific J Reprod 2016;5(2):111–115. [Google Scholar]
- 23.Kandeel A, Salim A, Zaghloul K.. Protective effect of L-carnitine against reproductive toxicity of monosodium glutamate in male albino mice. BMJ Br Med J 2019;6:10–32. [Google Scholar]
- 24.Casarini L, Crépieux P, Reiter E, et al. FSH for the treatment of male infertility. Int J Mol Sci 2020;21(7):1–20. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 25.Abu-Taweel GM. Effect of monosodium glutamate and aspartame on behavioral and biochemical parameters of male albino mice. African J Biotechnol 2016;15(15):601–612. [Google Scholar]
- 26.Ryu JH, Kang D.. Physicochemical properties, biological activity, health benefits, and general limitations of aged black garlic: A review. Molecules 2017;22(6):919. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 27.Chen M, Hao J, Yang Q, et al. Effects of icariin on reproductive functions in male rats. Molecules 2014;19(7):9502–9514. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 28.Zeid EHA, Alam RTM, El-Hameed NEA.. Impact of titanium dioxide on androgen receptors, seminal vesicles and thyroid hormones of male rats: Possible protective trial with aged garlic extract. Andrologia 2017;49(5): 10.1111/and.12651. [DOI] [PubMed] [Google Scholar]
- 29.Nasr AY, Alshali RA. Cytoprotective and antioxidant effects of aged garlic extract against adriamycin-induced cardiotoxicity in adult male rats. Anat Cell Biol 2020;53(2):201–215. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 30.Hajiuon B. Effects of garlic (Allium sativum L.) hydroalcoholic extract on estrogen, progesterone and testosterone levels in rats exposed to cell phone radiation. Zahedan J Res Med Sci 2014;16(12):20–25. [Google Scholar]
- 31.Dutta S, Sengupta P, Slama P, et al. Oxidative stress, testicular inflammatory pathways, and male reproduction. Int J Mol Sci 2021;22(18):1–20. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 32.Dutta S, Majzoub A, Agarwal A. Oxidative stress and sperm function: A systematic review on evaluation and management. Arab J Urol 2019;17(2):87–97. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 33.Mulyani SSTE, Adriani M, Wirjatmadi B.. Effect of mung bean sprouts extract to the morphology and motility spermatozoa in mice exposed monosodium glutamate. Int J Prev Public Heal Sci 2016;2(3):14–17. [Google Scholar]
- 34.Amić D, Stepanić V, Lučić B, et al. PM6 study of free radical scavenging mechanisms of flavonoids: Why does O-H bond dissociation enthalpy effectively represent free radical scavenging activity? J Mol Model 2013;19(6):2593–2603. [DOI] [PubMed] [Google Scholar]
- 35.Nasr N, Elmadawy M, Almadaly E, et al. Garlic powder attenuates apoptosis associated with lead acetate-induced testicular damage in adult male rats. Alexandria J Vet Sci 2017;54(1):70. [Google Scholar]
- 36.Abdennour C, Ouarda A.. Evaluation of the therapeutic efficiency of raw garlic on reproduction of domestic rabbits under lead induced toxicity. Ann Biol Res 2011;2(3):389–393. [Google Scholar]