Effect of co-administration of gallic acid and quercetin or gallic acid and rutin on impaired spermatogenesis and oxidative damage in a busulfan-treated rat model.

Gallic acid (GAL), rutin (RUT), and quercetin (QUE) are common antioxidant agents in fruits and vegetables with intriguing pharmacological effects. In the present study, we compared the therapeutic outcomes of GAL + QUE in comparison with GAL + RUT co-treatment in a busulfan (BUS) model of testicular injury in Wistar rats. BUS (4 mg kg-1 body weight (b.w) was injected intraperitoneally daily for 4 days. GAL + RUT or GAL + QUE (20 mg kg-1 b. w) was delivered by oral gavage for 52 days. Examination of the testes of BUS-treated rats both biochemically and under light microscopy revealed an increased level of lipid peroxidation, DNA fragmentation, glutathione-S-transferase, lactate dehydrogenase, gamma-glutamyl transpeptidase, alkaline phosphatase and acid phosphatase with a concomitant decrease in the level of antioxidants: glutathione, ascorbic acid, superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase activities, suggesting testicular injury. Tissue sections confirmed the testicular injury-induced by BUS, including diminished spermatogenesis score index, tubular diameter, gonado-somatic index, testis weight, epithelia thickness and higher percentage of aberrant tubules. GAL + QUE co-administration had better recovery effects than GAL + RUT on the biochemical markers and protected against BUS-induced testicular damage. GAL + QUE treatment regimen has better capacity to maintain the antioxidant capacity of the testes and is more potent at reducing BUS-induced oxidative damage compared to GAL + RUT.

Ascorbic acid attenuates gasoline-induced testicular toxicity, sperm quality deterioration, and testosterone imbalance in rats.

The present study evaluated the protective effect of ascorbic acid (ASCB) against gasoline fumes (PET) induced testicular oxidative stress, sperm toxicity, and testosterone imbalance in Wistar rats. Twenty-four (24) male albino rats (75 ± 16 g) were randomized into three experimental groups (N = 8). The control group: received normal saline, PET group: exposed to PET 6 h daily by inhalation in an exposure chamber and PET + 200 mg ASCB/kg body weight group: exposed to PET 6 h daily by inhalation and administered ASCB per os. Treatment of ASCB and PET exposure was done thrice and five times weekly for a period of 10 weeks respectively. ASCB co-treatment prevented PET-induced increases in the oxidative stress markers (glutathione, glutathione S-transferase, superoxide dismutase, catalase, hydrogen peroxide generation, nitric oxide, and lipid peroxidation) and serum testosterone concentration (p < .05). Sperm quality was low and those with damaged heads and tails increased alongside histological injuries in the PET-exposed rats, which were also minimized with ASCB administration. ASCB protected against PET-induced oxidative stress, sperm, and testis damage in rats.

Influence of triazines and lipopolysaccharide coexposure on inflammatory response and histopathological changes in the testis and liver of BalB/c mice.

Background: Triazines are environmental active chemicals that have been reported to alter the inflammatory status of the gonads. We tested the anti-inflammatory effect of the triazines (atrazine; ATZ, simazine; SMZ and cyanazine; CYZ) on the testis and compared it with the more classical liver model that has substantial populations of resident macrophages comparable to the testis. Methods: BalB/c mice were treated with 25 mg/kg ATZ, SMZ and CYZ for 30 days and injected with lipopolysaccharide (0.5 mg/kg i.p.) 6 h before sacrifice. Myeloperoxidase activity and nitric oxide level in the testis and liver homogenates were determined by spectrophotometry whereas tumor necrosis factor-alpha and interleukin-6 concentrations were evaluated by immunoassay. Haematoxylin and eosin stained sections of the tissues were observed using a light microscope. Results: Myeloperoxidase activity, nitric oxide, tumor necrosis factor-alpha, and interleukin-6 levels were decreased in the liver and testis of the triazines co-treated animals. SMZ has the most potent inhibitory effect and ATZ the least effect on inflammatory mediators in both tissues. Microscopic evaluation showed loss of inflammatory cells in the inter-tubular areas of the testis and few patchy masses of infiltrating inflammatory cells around the central vein of the liver. Conclusion: Triazines inhibit the levels of inflammatory mediators in the testis and liver of mice. The anti-inflammatory effect of triazines in a lipopolysaccharide-induced inflammation model was established in this study.

Testosterone restores TM3 and TM4 cell viability, reduces reactive oxygen species generation, and protects against atrazine-induced stereological changes in rat testes.

In this study, we performed the stereological examination of rat testes and evaluated the protective effect of testosterone against atrazine (ATZ) toxicity in TM3 Leydig and TM4 Sertoli cells. Testosterone intake in rats increased the volumetric density of the seminiferous tubules; tubular diameter; germinal epithelial height; number of spermatogonia, primary and secondary spermatocytes, round spermatids, Sertoli cells, and Leydig cells; and Johnsen scores compared with the values after ATZ treatment (p < 0.05). Furthermore, testosterone increased the viability of TM3 cells and reduced reactive oxygen species (ROS) generation in TM4 cells compared to the ATZ-treated group. In conclusion, exogenous testosterone intake maintains testicular morphometry and spermatogenesis in rats, and minimizes cell death and ROS generation in testicular cell lines exposed to ATZ. However, TM4 cells are more responsive to testosterone-mediated regulation of ROS generation induced by ATZ than TM3 cells.

Atrazine: cytotoxicity, oxidative stress, apoptosis, testicular effects and chemopreventive Interventions.

Atrazine (ATZ) is an environmental pollutant that interferes with several aspects of mammalian cellular processes including germ cell development, immunological, reproductive and neurological functions. At the level of human exposure, ATZ reduces sperm count and contribute to infertility in men. ATZ also induces morphological changes similar to apoptosis and initiates mitochondria-dependent cell death in several experimental models. When in vitro experimental models are exposed to ATZ, they are faced with increased levels of reactive oxygen species (ROS), cytotoxicity and decreased growth rate at dosages that may vary with cell types. This results in differing cytotoxic responses that are influenced by the nature of target cells, assay types and concentrations of ATZ. However, oxidative stress could play salient role in the observed cellular and genetic toxicity and apoptosis-like effects which could be abrogated by antioxidant vitamins and flavonoids, including vitamin E, quercetin, kolaviron, myricetin and bioactive extractives with antioxidant effects. This review focuses on the differential responses of cell types to ATZ toxicity, testicular effects of ATZ in both in vitro and in vivo models and chemopreventive strategies, so as to highlight the current state of the art on the toxicological outcomes of ATZ exposure in several experimental model systems.

Fluted pumpkin seeds protect the spermatogenesis score index and testicular histology of caffeine treated rats.

The present study was designed to evaluate the protective effect of fluted pumpkin seeds (FPS) against caffeine (CAFF) induced testicular toxicity in rats. Thirty young healthy male Wistar rats (196 ± 12 g) were randomly organized into five sets of six animals per each group: control, caffeine (CAFF; 50 mg kg-1 bw) and FPS co-treatment groups (CAFF + 50 mg FPS, CAFF + 100 mg FPS and CAFF + 200 mg FPS kg-1  bw). CAFF and FPS were administered daily and twice per week respectively by oral gavage for 40 days. CAFF treatment decreased testicular lactate dehydrogenase enzyme activity level, which was attenuated on co-administration with FPS at 50 and 100 mg kg-1  bw. Furthermore, CAFF decreased seminiferous epithelia thickness and spermatogenesis score index and increased the number of tubules with abnormal histological features, which were attenuated on co-administration with FPS at 50 mg kg-1  bw much more than at the higher doses (p < 0.05). CAFF did not affect malondialdehyde and glutathione concentrations and glutathione peroxidase (GSH-Px) activity in the testes whereas FPS co-treatment at the higher doses elevated glutathione level and GSH-Px activity and did not affect spermatogenesis score index at the highest dose (200 mg kg-1  bw). Testicular malondialdehyde concentrations remained unaffected in all FPS co-treatment groups. Overall, FPS is able to minimize the CAFF-induced testicular injury at lower than at the higher tested doses.

Oral antiviral treatments for COVID-19: opportunities and challenges.

The use of antiviral COVID-19 medications can successfully inhibit SARS-CoV-2 replication and prevent disease progression to a more severe form. However, the timing of antiviral treatment plays a crucial role in this regard. Oral antiviral drugs provide an opportunity to manage SARS-CoV-2 infection without a need for hospital admission, easing the general burden that COVID-19 can have on the healthcare system. This review paper (i) presents the potential pharmaceutical antiviral targets, including various host-based targets and viral-based targets, (ii) characterizes the first-generation anti-SARS-CoV-2 oral drugs (nirmatrelvir/ritonavir and molnupiravir), (iii) summarizes the clinical progress of other oral antivirals for use in COVID-19, (iv) discusses ethical issues in such clinical trials and (v) presents challenges associated with the use of oral antivirals in clinical practice. Oral COVID-19 antivirals represent a part of the strategy to adapt to long-term co-existence with SARS-CoV-2 in a manner that prevents healthcare from being overwhelmed. It is pivotal to ensure equal and fair global access to the currently available oral antivirals and those authorized in the future.

Curcumin improves the protective effects of quercetin against atrazine-induced testicular injury in adult Wistar rats.

This study evaluated the beneficial protective effect of cotreatment of curcumin (CUR) and quercetin (QUE) on atrazine (ATZ)-induced testicular toxicity in rats. ATZ challenge diminished luteinizing hormone, follicular stimulating hormone, testosterone and myeloperoxidase enzyme activity, but these effects were attenuated on co-treatment with CUR and QUE. Also, co-treatment of CUR + QUE was better than separate administration of QUE at diminishing malondialdehyde and glutathione and improving tumour necrosis factor-α concentration, germ cell numbers (spermatogonia, spermatocytes and round spermatids) and epididymal sperm quality. Histologically, smaller sized tubules with degenerated epithelia and few germ cells were seen in the seminiferous tubules of the ATZ group whereas CUR + QUE pretreatment improved the histo-morphologic features of the tubules compared to the ATZ group and was also better than separate administration of QUE. We conclude that CUR can improve the protective effects of QUE against ATZ-induced testicular injury by enhancing the levels of reproductive hormones, recovering testicular biochemical parameters and improving the histological features of the testes.

The protective effect of fluted pumpkin seeds against atrazine-induced testicular injury.

The present study evaluated the protective effect of fluted pumpkin seeds (FPS) on atrazine (ATZ)-induced testicular damage. Twenty adult male Wistar rats were divided into four groups of five animals each. The control animals (Group A) received corn oil (2 mL/kg body weight), Group B animals received 50 mg ATZ/kg body weight by gavage, while Groups C and D received 50 mg ATZ/kg body weight once a week in addition to the gavage of 25 mg FPS/kg body weight and 50 mg FPS/kg body weight respectively for 60 days. The results showed that testicular myeloperoxidase activity and nitrite concentration were decreased in all groups compared to the control value. The increase in malondialdehyde and decrease in glutathione concentrations in group B were abrogated in group C (p < 0.05) but not in group D animals. The increase in γ-glutamyl transpeptidase and decrease in lactate dehydrogenase enzyme activities in the group B animals were also normalized to control values in group C but not in group D animals. Interestingly, the testis of the group D animals showed massive depletion of germ cells and the diameter of the seminiferous tubules of these animals were decreased compared to all other groups. However, the number of motile sperms, abnormal sperms and sperm count in Group D animals were similar to the ATZ-treated animals and lower than control values. In conclusion, FPS protected against ATZ-induced testicular damage but can also harm the testis at a higher dose.

COVID-19 vaccinations: The unknowns, challenges, and hopes.

The entire world has been suffering from the coronavirus disease 2019 (COVID-19) pandemic since March 11, 2020. More than a year later, the COVID-19 vaccination brought hope to control this viral pandemic. Here, we review the unknowns of the COVID-19 vaccination, such as its longevity, asymptomatic spread, long-term side effects, and its efficacy on immunocompromised patients. In addition, we discuss challenges associated with the COVID-19 vaccination, such as the global access and distribution of vaccine doses, adherence to hygiene guidelines after vaccination, the emergence of novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, and vaccine resistance. Despite all these challenges and the fact that the end of the COVID-19 pandemic is still unclear, vaccines have brought great hope for the world, with several reports indicating a significant decline in the risk of COVID19-related infection and hospitalizations.

The protective effect of rutin against busulfan-induced testicular damage in adult rats.

Here, we studied the protective effect of rutin (RUT) against testicular damage caused by busulfan (BUS) in rats. Adult male Wistar rats were intraperitoneally injected with BUS (4 mg/kg body weight at day 7 and 14), and then treated with RUT (30 mg/kg body weight) by gavage thrice weekly for 60 days. The results showed that BUS-induced increase in 3ß-hydroxysteroid dehydrogenase (3ß-HSD) was significantly decreased by RUT, whereas 17ß-HSD activity and plasma testosterone concentration remained unaffected (p > 0.05). It was also observed that RUT inhibited BUS-induced increase in nitrite concentrations and myeloperoxidase enzyme activities in the plasma and testes (p < 0.05). Similarly, BUS-induced decrease in glutathione and increase in malondialdehyde concentrations in the testes were significantly normalized to control values by RUT. Finally, RUT administration showed some tendency to improve the architecture of the seminiferous epithelium of the rat's testes after BUS treatment. Overall, RUT inhibited BUS-induced oxidative damage and inflammation in the testis of an experimental rat model.

Gallic acid ameliorates busulfan-induced testicular toxicity and damage in mature rats.

Here, we studied the protective effect of gallic acid (GAL) as a potent anti-oxidant and anti-inflammatory agent against damage caused by busulfan (BUS) in the testes of adult rats. The adult Wistar rats were assigned as control, BUS: was intraperitoneally (i.p.) treated with busulfan (15 mg/kg, day 7 and 14), GAL + BUS: was co-treated with busulfan (i.p., 15 mg/kg, day 7 and 14) and orally treated (per os) with gallic acid (60 days, 20 mg/kg) and GAL: was treated with gallic acid (per os, 60 days, 20 mg/kg). The results showed that GAL co-treatment increased the numbers of spermatogonia (Type A and B), spermatocytes (primary and secondary) and round spermatids, along with the tubular diameter, epithelial height and gonado-somatic index. In addition, BUS-induced increase in 3ß-hydroxysteroid dehydrogenase and γ-glutamyl transpeptidase activities were inhibited on GAL co-treatment. Similarly, BUS-induced decrease in gluthathione concentration, catalase and superoxide dismutase activities along with increase in myeloperoxidase activity and malondialdehyde concentration were significantly normalized to control values on GAL co-treatment. Busulfan-induced elimination of tubular germ cells was completely prevented by GAL. Overall, GAL may inhibit BUS-mediated spermatogenesis arrest via decreasing inflammatory-mediated oxidative stress in a rat experimental model.

Antioxidant and anti-inflammatory protective effects of rutin and kolaviron against busulfan-induced testicular injuries in rats.

There are few treatment options, including the use of natural phenolics-based combination therapy for mitigating male infertility conditions associated with chemotherapy. Busulfan is an anti-cancer drug that leads to testicular problems in humans. Here, we studied the effect of co-treatment of rutin and kolaviron against busulfan-induced testis damage. Young adult male Wistar rats were intraperitoneally injected busulfan (4 mg/kg b.w), and then orally administered rutin (30 mg/kg b.w), and kolaviron (50 mg/kg b.w) alone and combined for 60 days. Results revealed that rutin and kolaviron alone or in combination reversed busulfan-induced increase in oxidative stress along with sperm quality of treated animals. However, kolaviron and rutin separately improved the concentrations of MDA and GSH and sperm quality more than when they were combined. Similarly, rutin and kolaviron separately or in combination preserved spermatogenesis and relieved busulfan-induced increase in nitric oxide concentration, myeloperoxidase and 3ß-hydroxysteroid dehydrogenase activities. Co-supplementation with kolaviron but not rutin nor when rutin was combined with kolaviron also improved the testicular level of tumor necrosis-alpha. Finally, the histological features in the testes caused by busulfan were reversed by rutin, whereas treatment with kolaviron alone or in combination with rutin partially protected the testis from busulfan-induced injury as demonstrated by the appearance of few germ cells, damaged tubules, loss of round spermatids and defoliation of the seminiferous epithelium. Thus, the combined treatment regimen of rutin and kolaviron sparingly prevented busulfan-induced testicular injuries in rats.Abbreviations: CAT: Catalase; GSH: Glutathione; 3ß-HSD: 3ß- hydroxysteroid Dehydrogenase; MDA: Malondialdehyde; TNF-α: Tumor necrosis-alpha; BUS: Busulfan; RUT: Rutin; KV: Kolaviron; TBARS: Thiobarbituric Acid Reactive Substances; MPO: Myeloperoxidase; ELISA: Enzyme-Linked Immunoassay; NAD: Nicotinamide Adenine Dinucleotide (oxidized); ROS: Reactive Oxygen Species.

Atrazine impairs testicular function in BalB/c mice by affecting Leydig cells.

Several studies have reported the effects of atrazine on the gonads of many experimental models. However, the short-term effects of in vivo exposure to atrazine on the testes of mice are not well clarified. Here we reported that adult BalB/c mice exposed to atrazine (50 mg kg-1 body weight) by gavage for three consecutive days have reduced numbers of 3ß-hydroxysteroid dehydrogenase positive Leydig cells (LCs), associated with increased in situ cell death fluorescence and caspase-3 immuno-expression in the testes. Consequently, immunostaining for cell cycle gene regulators showed increased expressions of p45, accompanied with increased expressions of cyclin D2 and E2. Histological observations of the gonads showed reduced number of germ cells in particular areas, sloughed seminiferous epithelium, presence of giant apoptotic cells close to the seminiferous tubule lumen and in the epididymal lumen along with low numbers of Leydig cells in the testicular interstitial areas. Similarly, LCs isolated from the testes of BalB/c mice that were exposed to atrazine (0.5, 25, 50 mg kg-1 body weight) in the same manner as in the first experiment presented dose-dependent increased caspase-3 activity, decreased cell viability, intratesticular and serum testosterone concentrations and LCs testosterone secretion. In summary, atrazine appears to directly decrease the number of testosterone secreting LCs in mice through apoptosis.

Antifertility and profertility effects of the leaves and seeds of fluted pumpkin: Sperm quality, hormonal effects and histomorphological changes in the testes of experimental animal models.

Fluted pumpkin (FP; Telfairia occidentalis) is an edible vegetable, grown in West Africa, that is used in traditional medicine for its regulatory effects on the male gonads. Scientific articles concerning the effects of FP were identified by searching PubMed, PubChem, Scopus, Springer, ResearchGate, Google Scholar and Web of Science; this literature was to better understand the effects of FP seed (FPS) and leaf (FPL) extracts on the testes. Data showed that in experimental animals extracts of FPL and FPS at 1/100 of the lethal dose promoted testis regeneration and improved testosterone concentration and sperm quality, while at higher doses they had antifertility effects. Several extracts of FPS and FPL, including ethanol, aqueous, methanol and hydroethanolic, had protective effects on the testes of study animals at lower doses (≥ 50 mg/kg body weight), but at higher doses (≥ 200 mg/kg body weight) they inhibited hormone synthesis, sperm quality and histomorphological structure, under both normal and disease conditions. The posttreatment effects of FPS on the gonads were reversible in young mature rats and FPS had slight systemic toxic effects. Although, there are inconsistencies in some of the published results, the current evidence suggests that FPS and FPL have both profertility and reversible antifertility effects in experimental animals.

Evaluation of the protective effects of quercetin and gallic acid against oxidative toxicity in rat's kidney and HEK-293 cells.

Quercetin and gallic acid are phytochemicals with interesting pharmacological properties. We herein investigated the protective effect of quercetin (QUE) in comparison with gallic acid (GAL) against exogenously-induced oxidative damage in rats' kidney and human embryonic kidney (HEK-293) cell lines. Adult Wistar rats were treated with QUE and GAL (50 mg/kg) separately or in combination with di-n-butylphthalate (DnBP) for 14 days; and HEK-293 cells were treated with different concentrations of GAL (25-294 µM) or QUE (2-17 µM or 28-165.43 µM) singly or in combination with H2O2 (200 µM). After treatment, the kidney and cell extracts were processed for biochemical analysis and histopathology. We found that GAL but not QUE prevented DnBP-induced increase in lipid peroxidation (2.603 ± 0.25 vs. 3.65 ± 0.21 µmol/mL). Treatment with QUE but not GAL was associated with increased plasma creatinine (729.09 ± 55.68 vs. 344.25 ± 50.78 µmol/l) and tissue malondialdehyde (3.72 ± 0.62 vs. 1.67 ± 0.47 µmol/mL) concentrations, along with histo-pathological changes such as glomerular and tubular degenerations. However, QUE exhibited wider therapeutic concentration ranges than GAL at which it inhibits lipid peroxidation in HEK-293 cells, and was found to inhibit H2O2-induced lipid peroxidation even at the lowest concentration (2 µM) that was tested (0.607 ± 0.074 vs. 0.927 ± 0.106 µmol/l). These suggest that the in vivo dosages required for the antioxidant protective effects of QUE in renal tissues are low.

Effect of Anthocleista djalonensis A. Chev Root Extract on Testis of Matured Rats.

OBJECTIVE: To investigate the effects of different extracts of Anthocleista djalonensis on the testis and epididymal sperms of rats. METHODS: Fifty male Wistar rats were randomly divided into 10 groups (n=5 in each group) and orally treated with 50, 100 and 200 mg/kg body weight each of methanol, aqueous ethanol (H-EtOH) and chloroform extracts of A. djalonensis. Corn oil was used as vehicle (2 mL/kg). After 60 days of treatment, testosterone (T) and cholesterol (CHOL) concentrations, catalase (CAT), lactate dehydrogenase (LDH), 3ß-hydroxysteroid dehydrogenase (3ß-HSD) and superoxide dismutase (SOD) activities in the testes along with myeloperoxidase (MPO) activities and nitrite concentrations (NO) in the serum and testes as well as sperm quality were measured. RESULTS: T and CHOL concentrations along with 3ß-HSD activity were significantly higher in the animals treated with the low dose than in those treated with the high dose of the chloroform extract (P<0.05). Furthermore, the chloroform extract was more effective than the methanol extract that had the most marginal effect on T level at the high dose and the H-EtOH extract that was only effective at the medium dose. LDH activity was dose-dependently increased by the extracts in all groups. The CAT-SOD antioxidant system was increased in the treated animals at all doses compared to the control values, but the increase in glutathione level reached significant level in those treated with the low dose H-EtOH aqueous ethanol extract (P<0.05). Only the high dose of chloroform extract had significant inhibitory effects on MPO activity (P<0.05). Serum NO concentration was decreased at all doses of the extracts. The inhibitory effects of the extracts on testicular NO concentrations follow this order, chloroform extract > H-EtOH > methanol. Although all extracts at all doses showed excellent stimulatory effects on sperm quality (count, motility and morphology), the methanol extract at the high dose was the most effective on sperm count (P<0.05). CONCLUSION: The chloroform extract of A. djalonensis has better androgen-like and anti-inflammatory effects whereas the methanol extract has the best effect on sperm count.

Morphometric Evaluation of the Seminiferous Tubules and the Antioxidant Protective Effects of Gallic Acid and Quercetin in the Testis and Liver of Butyl Phthalate Treated Rats.

The antioxidant protective effects of gallic acid (GAL) and quercetin (QUE) against oxidative stress induced by di-butyl phthalate (DnBP) in the liver and testis of rats were evaluated in this study. Adult albino Wistar rats (180-225 g) were treated with QUE or GAL (50 mg/kg) alone or in combination with DnBP (1 mL/kg) for 15 days. After treatment, tissue samples were taken for determination of glutathione and malondialdehyde levels, and superoxide dismutase and catalase activities. Serial sections of the testis and liver were stained with haematoxylin and eosin for microscopy and seminiferous tubular morphometry. As expected, DnBP induced oxidative stress was evident by increased malondialdehyde level in both organs. Co-treatment with GAL or QUE reversed the malondialdehyde by 45.42, 37.44 and 37.57%, 23.32% and catalase by 52.21, 70.15 and 85%, 38.14% in the testis and liver respectively whereas superoxide dismutase activity and glutathione level were differently modulated parallel to histopathological improvement in both tissues. The seminiferous tubular diameter, epithelial height, epithelial germ cell count and tubular length were significantly decreased by 11.09, 51.91, 40.65 and 11.10% respectively versus control values after DnBP treatments and were attenuated on co-treatment with GAL or QUE. Co-treatment with GAL afforded better protective effects in both tissues but QUE treatment alone appeared more effective than GAL on the investigated morphometric data. It seems likely that GAL or QUE prevented the tissue damage but the antioxidant profiles of the liver and testis are different in response to the oxidative stress.