Impact of Aqueous Extract of the Stem Bark of Anthocleista schweinfurthii Gilg (Loganiaceae) on Some Parameters of the Reproductive Function of Adult Albino Male Rats

Impact of


Introduction
Today, living beings suffer from various diseases, with drugs being the preferred first treatment option. Human beings have always believed that drugs have the capacity to regulate diseases and have devoted the field of Medicine to ensuring the health and well-being of patients [1]. Multiple treatment options exist, for the treatment of various diseases. For a specific disease, there is the best treatment option [2]. For example, antibiotics are used to cure infections [3]; glucocorticoids are widely used in the treatment of chronic diseases because of their immunosuppressive and anti-inflammatory properties [4]. However, the clinical efficacy of certain medicines or drugs is compromised by the metabolic effects of longterm or high dose treatment and their potential for negatively affecting other body functions. This is shown in the case of dexamethasone, a glucocorticoid; high doses are known to cause a spectrum of clinical features such as obesity, insulin resistance and glucose intolerance [5] and its long-term use is associated with numerous side effects, such as the development of osteoporosis [6]. It is also known that estrogens and phytoestrogens act like anthropogenic endocrine disrupting compounds [7] and interfere with spermatogenesis and reduce sperm quality and production [8]. Anthocleista schweinfurthii, a Loganiaceae, named "Yütrùm" in Foumban, a locality in the West Cameroon, is a shrub of secondary forests. It stems bark decoction is used empirically in the treatment of stomach aches, female infertility, hernias and ovarian problems [9]. Mezui and collaborators in 2015 showed that the extract of A. schweinfurthii possesses significant anti-ulcer activity [10]. The extract protects the gastric mucosa even when gastric acidity is high and accelerates the healing of chronic gastric ulcer symptoms. Also, the results of the work of Ngoungoure and collaborators in 2017 showed that A. schweinfurthii had estrogenic properties in ovariectomized rats by reducing certain postmenopausal symptoms [11]. Consequently, within the limits of knowledge till date, there is a lack of scientific data on the impact of this plant on reproductive function of male rats. The purpose of this study was to investigate the impact of the aqueous extract of the stem bark of Anthocleista schweinfurthii Gilg (Loganiaceae) on reproductive function of adult albino male rats.

Plant Material
Stem bark of A. schweinfurthii was collected at Foumban, Western region of Cameroon, in February 2020. Authentication of this plant was done in 2002 by Pr. ZAPFACK Louis, Botanist (systematic/ecology), Department of Vegetal Biology, University of Yaoundé I, Cameroon. The sample of the plant was identified at the national herbarium under the voucher number 9890/SRFCam.

Preparation of Extract
Stem barks were cleaned, cut in small pieces and dried at ambient temperature. Dried pieces of barks of A. schweinfurthii were pulverized and 160 g were added in 4 L of tap water and were boiled for 45 min. The decoction was decanted, filtered with Watmann N° 3 filter paper and the filtrate evaporated in dry oven at 45°C and a powder of 17 g was obtained for an extraction yield of 10.62%.

Animal Material
Male Wistar rats aged approximately 2 months and weighting about 150 g were obtained from Animal House of Laboratory of Animal Physiology of University of Yaoundé I. They were housed in plastic cages at room temperature under natural day/night cycle. They had free access to a standard soy-free rat diet and tap water. All the experimental protocol was undertaken in accordance with the guidelines established by the European Union on Animal Care (CEE Council 86/609) adopted by the Ethical Committee of the Cameroonian Ministry of Scientific Research and Technology Innovation (Ref. No FWA-IRD0001954).

Experimental Design
Male rats were randomly divided into three groups (A to C) comprising of six rats each. The group A (Nor + H 2 O) served as the control while B (Nor +AS 300) and C (Nor +AS 400) constituted the treated groups. The extract was prepared in distilled water and orally administered to treated groups at doses of 300 and 400 mg/kg/day respectively, while distilled water (1 mL/100 g/day) was given to the control groups for 28 days. Body weights were recorded weekly throughout the treatment period and before sacrifice.

Serum Collection
Sacrifice of rats under anaesthesia was carried on the 29th day, the next day after the termination of the whole experiments following over-night fasting of the animals. After sacrifice, blood samples were collected from the jugular vessels into clean and dry centrifugation tubes. The blood samples were centrifuged at 3000 rpm for 15 minutes and different sera were aspirated with pipettes into clean and dry Eppendorf tubes and stored at a temperature at -20°C for further biochemical analysis (proteins, cholesterol, LH, FSH, testosterone).

Tissues Collection
Thereafter, the rats were quickly dissected. The testis, the seminal vesicles and the epididymis were taken and rinsed with physiological solution (NaCl 0.9%). These organs were cleaned of superficial fatty layer and weighed using 4-digital electronic balance (Mettler PL301) for the determination of the testis/body, seminal vesicles/body and epididymis/body weight ratio. A part of the testis, epididymis and seminal vesicles were fixed in Bouinʼs fluid for histology investigations and other parts were used for preparation of homogenates.

Epididymal Sperm Motility, Viability and Counts
Epididymal sperm counts were performed to assess the rats' reproductive status. Shortly after dissection, about 6 mm length of the caudal epididymal tissue was cut into small pieces in a stemmed glass containing 10 mL of NaCl 0.9% solution and incubated in a water bath at 34°C temperature. This sperm was further used for determination of some sperm characteristics:

(i) Sperm Count and Mobility
A volume of 20 µl of the sperm suspension was aspirated and deposited on a Malassez cell. It was observed in a photonic microscope (OLYMPUS JAPAN), X 400 and the number of spermatozoa (mobile and immobile) was rapidly counted in five randomly selected quadrants [17]. Quota of mobile spermatozoa was calculated using the following formula: % Mobility = (Mobile sperm/Total number of sperm) × 100 The number of spermatozoa per mL of sperm (N) was calculated using the following formula according to Sultan et al., 1982, [18].
Sperm count = X = sperm count in 4 randomly selected quadrants of the Malassez's cell df = dilution factor (20).

(ii) Sperm Viability
Sperm viability assessed from eosin staining that discriminate life sperm from dead sperm by staining cytoplasm of cell. A volume of 20 µL of the sperm sample previously obtained was placed on a microscopic slide and 10 µL of eosin 0.5% added to it and then covered with a slide and observed with light microscope X 400. A total of 100 sperm were counted within 2 minutes after the addition of the stain. Evaluation of live (unstained) and necrospermic (stained) spermatozoa were done with light microscopy according to the method described by Talebi and collaborators in 2007 [19].

Preparation of Homogenates
0.4 g of testis sample was ground in a mortar and homogenised at 20% in the phosphate sodium buffer, pH =7.4 on an ice tray. 0.4 g of epididymis was also ground in a mortar and homogenised in a phosphate potassium buffer, pH = 6.8, at 20% on an ice tray. A part of seminal vesicle tissue was ground in a mortar too and homogenised at 25% in distilled water on ambient temperature. Then, the homogenates were centrifuged for 25 min at 3,000 rpm maintaining the temperature at 4°C. The supernatant obtained was assayed and stored at -20°C for the estimation of both biochemical analysis including oxidant stress markers (CAT, SOD, GSH, NO, MDA), total protein, testicular cholesterol and seminal fructose.

Biochemical Analysis
Total protein levels were determined using colorimetric methods described by Gornall and collaborators (1949) [20]. Assay kit was used to determine total cholesterol level. This kit was supplied by BIOLABO SAS, France.
The FSH SA ELISA and Testosterone ELISA kits were intended for the quantitative measurement of Follicle Stimulating Hormone (FSH) and testosterone respectively. These kits were supplied by CALBIOTECH, a life science company, El Cajon, California. The LH ELISA kit was used for quantitative determination of Luteinizing Hormone (LH). This kit was supplied by Dialab Company, Austria.
Fructose levels were determined in seminal vesicle following protocols described in a WHO manual [21].
Reduced glutathione (GSH) was assessed using the method described by Ellman, (1959) [22] and results were expressed as Mmol GSH/g protein. Lipid peroxidation levels in homogenates were estimated using the procedure described by Wilbur and collaborators (1949) [23] and results were expressed as Mmol MDA/g of organ. Superoxide dismutase (SOD) was assayed by following the method of Misra and Frisovich, (1978) [24]. The enzyme unit of activity was defined, as the enzyme required for 50% inhibition of epinephrine auto-oxidation. Catalase is a ferriheme-containing enzyme that is responsible for the conversion of hydrogen peroxide (but not other peroxides) to water [25]. This activity was measured by the method of Aebi (1974) and was expressed as mM of H 2 O 2 /min/g of protein [26]. Nitrites levels were assayed following the Griess reaction [27].

Histological Investigation
After measuring the weight of each organ sample for two weeks, a Bouin fixation for the testicular, seminal vesicle and epididymis tissues was performed to determine histological changes in the organs following treatment with distilled water or aqueous extract of A. schweinfurthii. Fixed tissues were dehydrated using a series of graded alcohol mixtures. The dehydrated tissue was then cleared in xylene for two hours and thirty minutes. Tissues were then embedded in paraffin and were cut at a thickness of 5 µm using a microtome (Reichert-Jung 2030). Sections of tissues were mounted on slides and stained by immersing them in Mayer hematoxylin solution. The slides were rinsed under running tap water to remove excess hematoxylin, dipped in alcohol, eosin solution and then dehydrated through a series of graded alcohols and cleared in xylene. Finally, the tissues were mounted under a synthetic resin. Microscopic evaluation of the slides was undertaken and variations in histo-architecture were recorded [28]. (Loganiaceae) on some Parameters of the Reproductive Function of Adult Albino Male Rats

Statistical Analysis
Statistical analysis was carried out using Graphpad Prism 8.0.1.244 software. Data are presented as mean ± standard error of mean (SEM). Results were assessed using one-way analysis of variance (ANOVA) followed by the Tukey posthoc test. A probability level less than 0.05 was accepted as significance.

Qualitative Phytochemical Analysis of the Aqueous Extract of Anthocleista Schweinfurthii
The qualitative phytochemical screening carried out on Anthocleista schweinfurthii stem bark aqueous extract revealed that it contained alkaloids, flavonoids, saponins, reduced sugars, tannins, phenols, steroids and triterpenes but not glycosides and lipids (Table 1).

Effect of Anthocleista Schweinfurthii Aqueous Extract on Percentage of Weight Gain
The graphic representation ( Figure 1) shows the results of weight gain. There was no significant diference in weight gain observed between extract-treated groups and their controls receiving distilled water.

Effects of Anthocleista Schweinfurthii Extract on Relative Weight of the Testes, the Epididymis and the Seminal Vesicles
The effects of A. schweinfurthii on the relative weight of the testes, the epididymis and the seminal vesicles are summarised in figure 2. The treatment with A. schweinffurthii aqueous extract during 28 days induced a dose dependent decreasing effect on certain sexual organs relative weight. We observed at doses of 300 mg/kg and 400 mg/kg, a significant lowering of the relative weight of testis (p < 0.005 and p < 0.001 respectively) and epididymis (P < 0.002 and p < 0,001 respectively) compared to the control group. Nevertheless, treatment with the plant extract for 28 days induced a decrease in seminal vesicle relative weight at the dose of 400 mg/kg. However, the reduction was not significant in comparison to their control group receiving placebo (distilled water). Data are shown as means ± SEM, n = 6/group. *p < 0,05; **p < 0,01; ***p < 0,001: statistically significant compared with Nor + H2O. Nor + H2O = normal animals treated with distilled water (control group), Nor + AS 300 and Nor + AS 400 = normal animals treated with A. schweinfurthii aqueous extract at the doses of 300 and 400 mg/kg respectively.

Effects of Anthocleista Schweinfurthii on Caudal Epididymis Sperm Count, Viability and Motility
Concerning semen analysis, it was found that oral administration of A. schweinfurthii extract at 300 mg/Kg and 400 mg/kg to adult male rats induced a significant decrease of the sperm count, sperm motility and viability ( Figure 3).

Epididymal Sperm Count
Epididymal sperm counts were decreased in A. schweinfurthii treated rats as compared to distilled water treated rats (control group). Indeed, the treatment with A. schweinfurthii at doses of 300 mg/kg and 400 mg/kg induced a significant decrease in number of caudal epididymal spermatozoa (p < 0.033 and p < 0.001 respectively) in relation to control group ( Figure 3A). Figure 3B shows the percentages of motile sperms from all groups. Rats that were treated with A. schweinfurthii at the both doses presented a significantly lower percentage of motile sperms compared to control group (p < 0.001).

Sperm Viability
The administration of A. schweinfurthii stem bark aqueous extract induced a significant decrease in sperm viability of animals treated with the 300 mg/kg dose (p < 0.002) and with the 400 mg/kg dose (p < 0.001) compared to control group ( Figure 3C).

Effects of Anthocleista Schweinfurthii Extract on Total Proteins
The figure 4 below shows the effects of oral administration of A. schweinfurthii aqueous extract on total proteins concentration in the serum ( Figure 4A) and in androgendependent organs ( Figure 4B). The treatment with plant extract for 28 days at the doses of 300 mg/kg and 400 mg/kg induced a significant reduction (p ˂ 0.002 and p ˂ 0.001 respectively) in total proteins concentration in comparison to the control group receiving distilled water. Total protein concentrations in testis were significantly reduced (P < 0.001) in animals treated with plant extract at the dose of 300 mg/kg in comparison to control group while the dose of 400 mg/kg induced a significant decrease (p ˂ 0.002) in total protein concentration in seminal vesicles and in epididymis. Data are shown as means ± SEM, n = 6/group. *p < 0.05; **p < 0.01; ***p < 0.001: statistically significant compared with Nor + H2O. Nor + H2O = normal animals treated with distilled water (control group), Nor + AS 300 and Nor + AS 400 = normal animals treated with Anthocleista schweinfurthii aqueous extract at the doses of 300 and 400 mg/kg respectively. (Loganiaceae) on some Parameters of the Reproductive Function of Adult Albino Male Rats

Effects of Anthocleista Schweinfurthii Extract on Total Cholesterol
The effects of A. schweinfurthii extract are shown on figure 5. A. schweinfurthii at the dose of 400 mg/kg significantly reduced (p < 0.001) serum total cholesterol concentration relative to the control group ( Figure 5A). In contrast, rats that were treated with A. schweinfurthii at the both doses showed a significant increase of the testicular total cholesterol concentration as compared to distilled water treated animals (p < 0.001) ( Figure 5B).

Figure 5. Effects of Anthocleista schweinfurthii extract on total cholesterol.
Data are shown as means ± SEM, n = 6/group. ***p < 0.001: statistically significant compared with Nor + H2O. Nor + H2O = normal animals treated with distilled water (control group), Nor + AS 300 and Nor + AS 400 = normal animals treated with Anthocleista schweinfurthii aqueous extract at the doses of 300 and 400 mg/kg respectively.

Effects of Anthocleista Schweinfurthii Extract on Serum Gonadotrophin Hormones (LH, FSH) and Serum Testosterone Concentrations
Statistically significant differences were found in the serum concentrations of both LH or FSH or in serum testosterone between the control group treated with distilled water and the group of animals treated with A. schweinfurthii aqueous extract at the doses of 300 and 400 mg/kg respectively ( Figure 6). Indeed, A. schweinfurthii at both doses showed a significant decrease in serum LH concentration and in the serum FSH concentration (p < 0.001 for LH concentration and p ˂ 0.033 at 300 mg/kg and p ˂ 0.002 at 400 mg/kg for FSH) (Figure 6 A) compared to the control group. Also, the extract plant at the both doses significantly reduced serum testosterone concentration (p < 0.001) ( Figure 6B) in comparison to animals treated with distilled water. Data are shown as means ± SEM, n = 6/group. *p < 0.05; **p < 0.01; ***p < 0.001: statistically significant compared with Nor + H2O. Nor + H2O = normal animals treated with distilled water (control group), Nor + AS 300 and Nor + AS 400 = normal animals treated with Anthocleista schweinfurthii aqueous extract at the doses of 300 and 400 mg/kg respectively.

Effects of Anthocleista Schweinfurthii Extract on Vesicular Fructose Level
The aqueous extract of A. schweinfurthii caused a significant reduction of vesicular fructose in group Nor + AS 300 (p < 0.05) and in group Nor + AS 400 (p < 0.001) when compared to control group receiving distilled water (Figure 7). Data are shown as means ± SEM, n = 6/group. *p < 0.05; ***p < 0.001: statistically significant compared with Nor + H2O. Nor + H2O = normal animals treated with distilled water (control group), Nor + AS 300 and Nor + AS 400 = normal animals treated with Anthocleista schweinfurthii aqueous extract at the doses of 300 and 400 mg/kg respectively. Table 2 and Figure 8 summarise the results of A. schweinfurthii stem bark aqueous extract on some oxidative stress parameters in the testes, the seminal vesicles and the epididymis, notably superoxide dismutase (SOD) and catalase (CAT) activities, reduced glutathione (GSH) levels, nitrites (NO) content and malondialdehyde (MDA) levels.

Effects of Anthocleista Schweinfurthii Extract on Some Oxidative Stress Parameters
Our results showed that 28 days of oral administration of A. schweinfurthii to male rats significantly increased SOD activity in testes (p < 0.001) at the both doses as well as catalase activities in a dose-dependent manner (p ˂ 0.033 at 300 mg/kg and p ˂ 0.001 at 400 mg/kg) compared to control group. At the dose of 400 mg/kg, we observed a significant increase in GSH levels (p ˂ 0.033) in testes in comparison to animals treated with distilled water. We also observed that in the test group, the plant extracts significantly decreased testicular MDA levels, in a dose-dependent manner (p ˂ 0.033 at 300 mg/kg and p ˂ 0.002 at 400 mg/kg) compared to the control group.
Concerning seminal vesicle, it was found that oral administration of A. schweinfurthii induced a significant lowering of MDA levels (p ˂ 0.001) at all treated doses. Also, The 400 mg/kg dose of A. schweinfurthii significantly decreased CAT activity (p ˂ 0.033) comparatively to animals treated with distilled water. Meanwhile, no statistically significant differences were found in SOD activity, GSH and nitrites levels between normal animals treated with distilled water and normal animals treated with plant extract.
In the epididymis, Treatment with A. schweinfurthii extracts at the two doses significantly increased GSH (p ˂ 0.002) and nitrites (p ˂ 0.033 at 300 mg/kg and p ˂ 0.002 at 400 mg/kg) levels versus control group. Also, we observed a significant decrease in MDA levels (p ˂ 0.001) at the both doses compared to control group.

Figure 8. Effects of Anthocleista schweinfurthii extract on CAT (A) and SOD (B) in testes, seminal vesicle and epididymis.
Data are shown as means ± SEM, n = 6/group. *p < 0.05; ***p < 0.001: statistically significant compared with Nor + H2O. Nor + H2O = normal animals treated with distilled water (control group), Nor + AS 300 and Nor + AS 400 = normal animals treated with Anthocleista schweinfurthii aqueous extract at the doses of 300 and 400 mg/kg respectively.

NITRITES (mmol/g of organ) Mean (ST) Groups
T

Effects of Anthocleista Schweinfurthii Extract on the Histology of the Testis, the Seminal Vesicle and the Epididymis
Microphotography of the testis of rats who received distilled water showed normal tissue architecture, with a normal ongoing spermatogenesis process in the seminiferous tubules. Lumen was found to be full of spermatozoa. The testis of A. schweinfurthii treated groups presented a clearing of lumen of seminiferous tubules of in dose dependent manner, proof of an impairment of the spermatogenesis process ( Figure 9).  As shown in figure 10, the transversal section of epididymis of rats which received distilled water also exhibited a normal architecture with a lot of spermatozoa in the lumen. In the lumen of epididymal tubules of rats treated with distilled water, spermatozoa density decreased.
In the lumen of the seminal vesicle of animals treated with distilled water, we observed seminal secretions. On the other hand, there was no secretion in the lumen of seminal vesicle of animals treated with A. schweinfurthii at dose 400 mg/kg ( Figure 11).

Discussion
Herbal medicines have received greater attention as alternative to clinical therapy of many diseases and the demand for these remedies has currently increased [29], in spite of its minimal known "side-effects" on human health [30]. Number of studies have demonstrated the effect of the stem bark aqueous extract of A. schweinfurthi on acute and chronic gastric ulcer rats' models in rats and our previous studies [11] have shown its oestrogenic effect on some postmenopausal symptoms. This study assessed the impact of the aqueous extract of the stem bark of A. schweinfurthii on some parameters of the reproductive function of adult albino male rats.
The results obtained in this study showed that oral administration of A. schweinfurthi at doses of 300 mg/kg and 400 mg/kg for 28 days caused a significant decrease in the relative weight of the testis, the epididymis and the seminal vesicles compared to a control group. This is a reflection of the site of action of the flavonoids, a phytoestrogen compound present in aqueous extract of A. schweinfurthii, which tends to bind to both estrogen receptors types (α and β) present within the testis, epididymis and seminal vesicle [31,32]. The results are in accordance with the study of Gray and collaborators (2001) [33] which proved that substances which have an effect on androgens action can reduce testis, epididymis, seminal vesicles and prostate weight. Indeed, oestrogens and certain phytoestrogens tend to have 'demasculinising' or anti-androgenic effects which are known to interfere with androgens biosynthesis, or alter the number of androgens receptors [34][35][36]. The decrease in the relative organs weight observed following the administration of the plant extract might be attributed to decrease in rates of total proteins in testes, seminal vesicle and epididymis. In fact, according to Lee and Lee, (1996) [37] as well as Kouakou and Benie, (2003) [38], proteins are essentials components of cells. Therefore, its decrease could induce organ atrophy. Also, total proteins in testis and epididymis are required for spermatogenesis and sperm maturation [39]. Thus, the significant reduction in the concentration of testicular and epididymal proteins in extract plant-treated rats can lead to impaired sperm maturation. Another explanation for the decrease in relative organs' weight could be a significant reduction of sperm parameters (count, mobility and viability).
Sperm Count, motility and viability were significantly affected by A. schweinfurthii in treated groups. There was a drastic fall in the sperm count and reduction in percentages of progressively motile and viable spermatozoa in both dosetreated animals. In fact, a negative effect of molecules with estrogenic action-like compounds derived from plants, such as flavonoids, have been reported to directly affect male sexual functions via spermatogenesis, sperm capacitation [40][41][42]. It is known that A. schweinfurthii exhibited estrogenic properties because it contains flavonoids and phenols. A negative effect on spermatogenesis could be due to increasing apoptosis of developing germ cells [43] and then, a lower number of spermatozoa. Motility is one of the most important features of fertile spermatozoa, widely used as an indicator of sperm function [39]. So, the plant extract might slow down and/or inhibit the spermatogenesis process in the testis, deteriorating sperm quality in the extract-treated group. Our results suggest that A. schweinfurthii stem bark aqueous extract may have anti-fertility effects suggested by the drastic fall in the sperm count and reduction in percentages of progressively motile and viable spermatozoa. This anti-fertility effect of the extract plant is confirmed on microphotography of testis, epididymis and seminal secretion which showed seminiferous tubules lumen clearing, decreased lumen spermatozoa density in epididymal tubules and the absence of secretions in the lumen of seminal vesicles.
Cholesterol is a precursor of steroid hormones biosynthesis. In the present study, rats treated with A. schweinfurthii extract presented a significant decrease in serum total cholesterol and a significant increase in testicular cholesterol compared to control group. Several explanations are possible for significant decrease in serum total cholesterol in rats treated with A. schweinfurthii. One possibility is the ability of secondary metabolites of A. schweinfurthii to activate estrogen receptors and induce the transcription of genes involved in lipid metabolism [11]. Moreover, the aqueous extract of A. schweinfurthii contain saponins, which are known for their hypocholesterolemic properties [44]. This hypocholesterolemic effect may be due to the inhibition of cholesterol absorption in the intestine leading to an increase in fecal cholesterol excretion [45]. An increase in testicular cholesterol concentration may reflect reduced conversion of cholesterol to testosterone [46]. The lower concentration of testosterone is the result of estrogenic effect of our plant extract. The same result was obtained by Caceres and collaborators [47] and Corrêa and collaborators [48] respectively in 2014 and 2017 who reported that soybean phytoestrogens reduce testosterone serum concentrations.
In this study, we found a significant decrease in serum gonadotrophin hormones concentrations (LH, FSH). Indeed, Shittu and collaborators (2008) had reported that steroidogenesis and the hypothalamic-pituitary-gonadal axis may appear to be important loci for phytoestrogen actions in the brain [32]. Phytooestrogens are non-steroidal compounds that can bind to both estrogen receptor alpha and estrogen receptor beta because of their ability to mimic the conformational structure of estradiol [49] (Kuiper et al., 1998). Phytooestrogens are found in many vegetables. Thus, this result is due to the ability of estrogen or estrogen-like molecules to decrease the synthesis and/or release of gonadotrophins with the implication of both low serums [50].
Fructose is essential for spermatozoa metabolism and spermatozoa motility. Fructose is an energy source for spermatozoa [51]. Treatment with the aqueous extract at both doses decreased seminal fructose content. In fact, it has been reported by Kameni and collaborators in 2017 [8] that after ejaculation, the spermatozoa in a process named fructolysis, consume fructose. At higher sperm counts, the process is stronger resulting in a low seminal fructose concentration, suggesting that the high fructose concentrations observed in animals treated with distilled water, were released to supply motile sperm with energy after ejaculation. Another explanation for the decrease in seminal fructose is a decrease of LH concentration which affects normal function of seminal vesicles [52].
A. schweinfurthii exhibits an antioxidant property highlighted by a decrease in MDA concentration and an increase in CAT, SOD and GSH in testis, an increase of CAT and a decrease of MDA in seminal vesicle and a decrease of MDA and an increase of GSH and NO in epididymis. These results might be attributed to plant secondary metabolites flavonoids, saponins, alkaloids, phenols known for their significant antioxidant activities [53,54] through radical scavenging, inhibition of the production of reactive oxygen species or inhibition of lipid peroxidation [55].

Conclusion
We found that there is a decrease in the relative mass of the androgen-dependent organs, impaired spermatogenesis and sperm quality, decrease of serum LH, FSH and testosterone levels and also a decrease serum cholesterol and increased (Loganiaceae) on some Parameters of the Reproductive Function of Adult Albino Male Rats testicular cholesterol while the rats consume this plant.
From these findings, it is concluded that Anthocleista schweinfurthii stem bark alters some parameters of the reproductive function of adult albino male rats and suggest a limitation of their use in male subjects.

Research Perspectives
We want to determine the effects on the offspring following parental exposure to the plant extract and also to determine the effects of acute administration of the plant extract on specific periods of development.