The male reproductive system is very susceptible many factors including chemicals, environmental and industral pollutants as well as bad dietary [1]. The epididymis is an essential accessory organ in male reproductive system. It is important for sperms motality, fertilization and ability of development [2]. In animals, steroidal hormones are essential for both reproduction as well as the stress responses. The primary cause of infertility in males is their incapacity to generate an adequate quantity of viable, healthy sperm. So, one of the most crucial indicators of men's reproductive health is the ability of the testicles to operate normally. As the main sex hormone with testosterone is important in this sense and a low level of it can lead to infertility, libido loss  and muscle weakness. Ageing, diabetes, oxidative stress and hazardous materials are a few examples of internal or external variables that affect normal testicular function [3]. Sexual system is considered sensitive to environment and type of nutrition. These are enough to cause diseases then reducing fertility.

Athletic supplements are often utilized in athletic and bodybuilding circles for its potential to enhance muscle mass and athletic performance. Though much attention has been focused on its benefits for physical strength, there is less awareness about its impacts on sexual health, particularly in male rats. These animals are frequently used in scientific studies due to their physiological similarities to humans and well-documented sexual behaviors, making them ideal subjects for examining the sexual effects of supplements like proteins, creatine and carbohydrates.

Nonetheless, engaging in athletic competition might result in temporary harm to the system of reproduction as well as sexual dysfunction (genital discomfort, hypoesthesia of the genitalia, hypogonadism, DE, lowered sexual operate, etc.) or irreversible (hypogonadism, DE, etc.), through direct action (external sexual organs wounds, saddle-related diseases in cyclists, etc.) or indirect action (drug use, substance addiction, tension, exercise-related hypogonadism, etc.) [4].

The intersection of nutrition and male reproductive health has an increasing attention in recent years, particularly in relation to how dietary interventions can influence reproductive outcomes [5]. Provide a foundational understanding of the complexities surrounding diet and male reproduction, emphasizing the necessity for collaborative research between nutrition and reproductive biology. Different researches focus on that the timing and duration of dietary treatments can yield vastly different outcomes, especially when comparing pre-pubertal and sexually mature males. It is thought that the best age for experiments on rats reproductive system is 10 weeks and more, because it’s the age of puberty and all the reproductive organs functionally and structurely are perfect. Some scientist advocate for a multifaceted approach to assessing fertility, suggesting that no single assay can comprehensively measure reproductive success [6]. This nuanced perspective underscores the importance of recognizing sex-specific responses to dietary macronutrient ratios [7].

Pini et al. [5] were suggesting that male reproductive structure may be uniquely affected by specific dietary components. The insights from their research underscore the necessity of understanding the distinct nutritional needs and responses of male rats, particularly in the context of protein supplementation and its potential effects on reproductive health.

Increasing caloric intake by supplements used to combat excessive energy use which is in turn enough to reverse the functions of sexual system towards worse. supplementation taken by athletes usually taken for increasing the improvement of performance [8]. Certain supplements have high protein or creatine contents or carbohydrate depending on the purpose from that supplements. In an otherwise healthy person, their use may result in elevated blood urea or creatine. 

In the study of Crittenden et al. [9], underscore the significance of specific protein isolates in enhancing athletic performance, suggesting that elite athletes require higher protein intake for optimal muscular function. This research highlights that not all dietary proteins exert the same effects, with dairy proteins such as casein and whey demonstrating distinct physiological impacts post-ingestion. The implications of this study are critical as they lay the groundwork for understanding how protein supplementation might not only support athletic performance but also potentially influence hormonal levels and, by extension, reproductive health.

The effects of creatine supplementation on muscular performance have been extensively examined, but less is known about how it affects male rats' sexual systems. Based on available studies and experimental evidence, creatine supplementation does not directly affect reproductive hormones or the sexual system, although some findings suggest possible interactions with metabolism, oxidative stress and general physiology that could indirectly influence reproductive function.

Athletes have begun using creatine supplements because of its positive effects on performance. Its impact on sexual behavior is still debatable [10]. Prior research examining the impact of creatine on physical performance has shown possible behavioral effects, but sexual health was rarely the main emphasis. This work supports previous research showing a rise in performance at first, followed by a fall at larger doses, indicating that this biphasic response might be typical for a variety of physiological domains [11].

In male rats, moderate creatine consumption seems to improve sexual behavior and performance; this is mostly shown in the form of enhanced desire, mounting and prolonged copulatory phases. These benefits go beyond physical athletics and are probably the consequence of increased muscle function and general energy reserves [12].

Despite the generally recognized usefulness of special diets for improving physical performance, the use of sports supplements may have a negative effect on the body. Carbohydrate supplements, frequently used by athletes to increase strength and physical endurance, not only lead to various disorders of the gastrointestinal tract but also impair the function of other organs and systems. 

The effects of the carbohydrate supplements maltodextrin and glucose administered to male rats during puberty on the dynamics of the blood steroid transport levels and the structure of the sexual system in pubertal and adult animals. The administration of carbohydrate supplements was found to be followed by changes in testosterone or certain sex steroid-binding globulin levels during puberty or in adult animals [13].

These studies provide a foundation for comprehending the complex interactions between male reproductive health and athletic supplements, especially when it comes to hormonal balance and sex organs histology. The body of research indicates that the three supplementation has consequences on male rat reproductive health towards worse. Therefore, more research is needed to fully understand these implications.

The exploration of how athletic supplements, like proteinic, creatine and hydrocarbonic supplements, influence male reproductive systems is an area with few researches that intertwines the fields of nutrition and reproductive biology. The literature reveals a progression of insights that elucidate the complex relationships between supplements intake, biochemical balance and reproductive health in male rat. 

Dosage and Administration

Twenty-four males of albino rats aged (11±4) weeks to ensure sexual maturity, physiological relevance with balancing consistency. Their weight rate were (133) gm were used in the experiment. The animels were divided into four groups each group contains sixrats, the feding is by gavage tube:

• Group (C): Dranched normal feed and distailed water

• Group (G1): Dranched concentration o.627 gm/mL of al-mass with carbohydrate nature

• Group(G2): Dranched creatine with concentration 0.7 gm/mL

• Group(G3): Dranched with isotop of whey protein nature with concentration0.636 gm/mL

The duration of the experiment were thirty days, then the animal were weighted and sacrificed for collecting serum and sexual organs.

BioChemical Test

At the end of the experiment, the animals were under anasthesia by inhealing chlorophorm then sacrificed for collecting blood serum and sexual organs. Serum samples were collected by using centerfuge 3000 pm/15 minutes three sex hormones (FSH, Tstosteron and LH) were estimated. By Using Enzyme-Linked Immunosorbent Assay (ELISA) kits (DiaSorin Inc., USA), this used the conventional sandwich enzyme-linked immune-sorbent assay methodology as its foundation.

Histophathological Preparation

As a part of ultimate protocol, sexual organs were isolated after sacrificing animals and adipose tissues were removed. The organs Samples were fixed by keping in formalin 10%, then dehydrated with ethanol, followed by cleaning with xylene. Then the samples embedded in paraffin as final step of preparation blocks of samples and creation of histological sections. After that, sample blocks are cut by using microtome. The section are put on slides and stained with hematoxiline and eosin stain. Then they examined by light microscope with power 10X and 40X [14].

Statistical Analysis

 The Statistical Analysis System- SAS (2018) program was used to detect the effect of difference factors in study parameters. Least significant difference–LSD test (Analysis of Variation-ANOVA) was used to significant compare between means in this study [15].

Results of Sexual Hormones Estimation

In Table 1, there are non-significant increasing levels of testosterone in both groups G1 and G2 by 5.89±4.59 IU/L in comparsion to control group ((5.23±2.64) IU/L, while G3 of has decreasing value of testosterone also non-significantly to (4.93±2.88) IU/L in comparsion to other groups. 

The following table shows a non-significant increasing in Luteinizing Hormone (LH) in G2 and G3 with values (2.30±0.70) IU/L and (1.67±0.34) IU/L respectively higher than control group (1.43±0.57) IU/L and G1 (1.20±0.17) IU/L. 

Table 1: Comparison between Difference Groups in Sex Hormones (IU/Liter)

NS: Non-Signifacant

Figure 1: Horizantal Microscopic Section for Male Rats, Shows Cauda of Normal Epididymal Tubules Filled with Sperm (Double Arrow) and there is Normal Steriocillia (Arrow), (Control), H&E Stain, 10X.40X

Follicle Stimulated Hormone (FSH) rises in serum but non-significantly in both G1 and G3 to (2.61±1.38)IU/L and (2.63±0.96) IU/L respectively in comparsion to control group (1.43±0.96) IU/L, while G2 has nearly the same value (1.47±0.89) IU/L to control group.

Results of Histological Changes in Reproductive Organs

After sacrificing the animals of all treatment groups, male reproductive organsincluding epididymis, seminal vesicles and testis were prepared for making histological sections and stained by hematoxlin and eosin staining. The final sections were examined microscopally by using two powers 40x and 10x to search possible histopathological changes in organs architecture in comparsion to normal control group.

Epididymis 

Figure 1 shows normal epididymal tubules of control group. It is filled with sperms and the inner edge contain normal steriocillia. The section shows normal diameter of tubules, but Figure 2 of the epididymal tubules for male rats dranched carbohydtratic supplement showed different diameters. Most of them has few steriocillia in the inner edge. The luminal of tubules either contain fewer sperms than usual or completely empty of sperms. There is also seen a congestion and hemorridge among tubules. There is also infiltration of neutrophils in interstitium, necrosis in epithelial cells, In addition to few hyperplasia of clear cells in the lining of epididymal tubules. 

Figure 2: Horizantal Microscopic Section showed the Epididymal Tubules in Male Rats of G1. A-the Tubules with Abnormal different Diameters, Congestion (Arrowhead), Necrosis of Epthelial Cells (Arrow), Neutrophilic Infiltration in Interstitium (Yellow Arrowhead).  B-there is no Sperms in the Lumen (Arrow) and Few Steriocillia (Thin Arrow), Narrow Cell (NC), Dendritic Cell (DC), Basal Cell (BC), Principal Cell (PC), Hyperplasia of Clear Cells (Arrowhead). H&E Stain, 10X & 40X

In the same as In the Figure 3, where there is a clear effect of creatine on the structure of cauda epididumal tubules. The horizantal section shows necrotic area among tubules. There are tubules lumen are empty of sperms with fewer stereocilia. 

The most effect among groups is in when male rats dranched proteinic supplements. The effect as illustrated in Figure 4, there are much changes in epididymis like hemorrage, lumen empty of sperms, few stereocillia, necrotic area among lobules and hyperplasia of clear cells.

Seminal Vesicles

In comparssion to normal section of seminal vesicles in Figure 5 which appears normal architecture of seminal vesicles with normal amount of lubricating fluid, normal mucosal tertary and secondary fold lining with epithelial secretory cells. The three athletic supplements shows through Figures 6-8 that there are clear alterations through the reduction of seminal vesicles lubricating fluid. 

Figure 3: Horizantal Microscopic Section of Cauda Epididymal Tubules for Male Rats of G2, A-Shows Lumen in Some Epididymal Tubules are Empty of Sperms (Arrow L), Hemorridge (Arrow), Necrotic Area (Rounded Head). B-Empty Lumen  of Sperms (Thick Arrow) with Few Steriocilli (Arrowhead), Principal Cells (Strightline), Basal Cells (Douple Arrow), Connective Tissue (Arrow). H&E Stain, 10X & 40X

So, the lumen appears empty of fluid. The most effect can be seen in the treatments with carbohydrate and creatine in groups (G2 and G3). The alterations also involve thickening the layer of interstial smooth muscle. Hyperchromatic muscle cell nuclei can be seen in the section of Figure 6. degeneration in some areas are noticed in epithelial tissue within mucosal folds. When male rats treated with creatine, seminal vesicles contain regions of hemorrage, in addition to necrotic and atrophy areas within epthelial tissues. The smooth muscle layer has profuse fibers as well as congestions (Figure 7).

In Figure 8 of G3, the alteration in the architecture of seminal vesicles is seen in tall, columinal, pseudostratified epithelial cells. The alteration involves shrinking of epithelial cells.

Results of Testis

According to Figure 9 of control group, normal section of seminiferous tubules can be seen clearly. The seminiferous tubules has normal diameter and contain normal development of germ cells. Sertoli cells with extending cytoplasm are found within germ cells. Leydig cells are there among seminiferous tubules.

Figure 4: Horizantal Microscopic Section of Cauda Epididymis in G3, A-Showed Hemorrhage (Arrow), Necrotic Area (Arrow), Lumen Empty of Sperms (L). B-Showed Epidedymal Lobules with Few Steriocilli (Yellow Arrow), Lumen is Empty of Sperms (Thick Arrow), hyperplasia of Clear Cells (Arrow). H&E Stain, 10X & 40X

Figure 5: Horizantal Microscopic Section of Seminal Vesicles in Male Rats (Control). Showed Normal Stratified Epithelium Tissue (Arrow), Normal Smooth Muscle Tissue (Arrowhead), Lubricant Fluid (LF).  H&E stain, 10X

Figure 6: Horizantal Microscopic Section Showed the Seminal Vesicles in Male Rats of G1. Showed Few Lubricant Fluid (Arrow), Thick Intersitial Tissue (Arrowhead), Degeneration of Epithelial Tissue (Yellow Arrow), Hyperchromatic Muscle Cell Nuclei (Red Arrow). H&E Stain. 10X

Figure 7: Horizantal Microscopic Section Showed the Seminal Vesicles in Male Rats of G2. It Showed Empty from Secretion (Arrow), Necrotic Area (Yellow Arrow), Atrophy of Seminal Vesicles Tubules (Arrowhead), Thickening of Interstitial Tissue with Profuse Fiber (Yellow Head), Congestion (Red Arrow), Hemorragh (Sequare Arrow). H&E stain. 10X

Figure 8: Horizantal Microscopic Section Showed the Seminal Vesicles in Male Rats of G3. It Showed Some Lobules are Empty from Secrtion (Arrow), some Lobules with Scanty Lubricant Fluid (S), Hemorrage in Intersitial Tissue (Thick Arrow), Degeneration of Stratifies Epithelial tissue (arrowhead), necrosis (yelow arrow). H&E stain. 10X

Figure 9: Horizantal Microscopic Section Showed Testis in Male Rats (Control). A- Showed Normal Section of Seminipherous Tubules with Leydig Cells (L). B- Showed Normal Spermatogenesis with Normal Spermatogonia (SG), Normal Primary and Secondary Spermatocytes (SC), Spermatide (SD), Sperms (SM) and Spermatozoa (SZ) with Normal Steriocells (Arrow). H&E Stain. 10X & 40X.

Figure 10: Horizantal Microscopic Section Showed Testis in Male Rats of G1. A- Most Seminipherous Tubules Showed Few Sperms with Wide Lumen (Arrow). B- Showed Vaculation of Spermatogonia (arrowhead), Wide Lumen and Empty of Sepermatozoa (Arrow) (EM). H&E Stain. 10X & 40X

Figure 11: Horizantal Microscopic Section Showed Testis in Male Rats of G2. A- Most Seminipherous Tubules Showed Wide Lumen (Arrow). B- Showed Irregular Spermatogenesis (IR) with Vaculation of Spermatogonia (Arrow), Wide Lumen and Empty of Spermatozoa (Arrowhead), Large Number of Leydig Cells (Yellow Arrow). H&E Stain. 10X & 40X

Figure 12: Horizantal Microscopic Section Showed Testis in Male Rats G3. A- Most Seminipherous Tubules Showed Lumen (Arrow) and High Numbers of Leydig Cells Among Tubule (Arrowhead). B- Showed Vaculation of Spermatogonia (Arrow), Wide Lumen Empty of Spermatozoa (Arrowhead), Degeneration of Spermatogenesis (Yellow Arrow), Leydig Cells (Yellow Head). H&E Stain. 10X & 40X

In comparssion to normal male rats in control group, Testis of treatment group G1 involving rats dranched carbohydrate supplement, it is found that that the alteration is appeared clearly. The alteration involves different diameters of tubules in addition to the empty lumen from spermatozoa in most tubules. Vaculationare found among spermatids (Figure 10).

Testis of rats dranched creatine has less effected some of tubules are empty of spermatozoa. Spermatogenesis is defected in some states of irregularty. Leydig cells are concentrated with high number among tubules. Vaculation is present (Figure 11)

In Figure 12 of males treated with proteinic supplement, testis contain seminiferous tubules with different diameters. Vaculation is clear among spermatids. Lumen is empty from spermatozoa. There is a sort of degeneratio of spermatogenesis.

Micronutrients are used by many athletes to improve their performance. Because it has both ergogenic and anabolic benefits. creatine has grown to be one of the most popular supplements on the market, thencomes protein and carbohydrates [16].

Through the results above, it is noticed that carbohydrates and creatine supplements have an effects on testosterone hormone towards increasing. Testosterone is crucial for regulating male reproductive function, including libido and sperm production. The connections between supplementation and hormones can originate from the potential for random (or intentional) exposure to restricted substances (hormones or prohormones) that are not disclosed on the labels of various nonhormonal supplement companies (i.e., vitamins, creatine, amino acids…etc.) [17].

This is my related to several intrepetations. Their effects on testosterone levels in rats have been studied, but results can vary depending on several factors, including the dosage, duration of supplementation and the specific strains of rats used. Through the promotion of muscle development and recuperation. A high protein diet can indirectly affect testosterone levels. Because anabolic signaling is amplified when muscle mass increases. Some findings indicate that certain protein supplements might affect the Hypothalamic-Pituitary-Gonadal (HPG) axis, a key hormonal pathway involved in the sexual system. Disruption of this axis could lead to changes in testosterone levels, sperm quality and reproductive organ health.

Testosterone production may rise accordingly, this depends on the situation. Moderate amount of protein may induce oxidative stress, ultimately impairing reproductive function [18]. When male rats were given athletic protein, the mean concentration of testosterone hormone increased non-significantly. The impact of protein during Leydig cell stimulation in the testes and LH stimulation to produce testosterone in Leydig cells, which may then operate on Sertoli cells, may be the cause of the rise in testosterone concentration. Numerous research have enhanced the impact of supplements on the production of testosterone. Zamir [19] highlighted the notable rise in testosterone hormone in males who supplemented with vitamin D. Men's levels of the hormone testosterone increased significantly when they supplemented with BCAA proteins [20]. 

The results shows that the three supplements participate non-significantly in changing serum levels of LH. Protein supplement incresases LH level in serum. The only intepretation is Consuming protein can indirectly alter muscle growth and recovery, which in turn can affect hormone levels, including LH. This agree with finding of Khaleel and Haba [21] when they studied the effects of whey protien on some hormones in rats. This completely disagree with the study of Goodin [22] human where Males supplementing with soy protein powder showed noticeably lower LH concentrations. Because soy isoflavones have some estrogenic properties, it is hypothesized that soy may have an effect on the hypothalamicpituitary-gonadal axis, reducing androgen synthesis. But according to Rahman [23], study found that twelve athletes who used large quantities of whey protein supplements daily had lower levels of the Luteinizing Hormone (LH) before and after exercise, which may indicate a pituitary gland malfunction.

Increasing levels of testosterone, LH and FSH during puberty male rats lead to testicular maturity and secondary features. Secondary characteristics of males and the production of testosterone are regulated by the same hypothalamic-pituitary axis. GnRH increases the release of FSH as well as LH from the pituitary, which in turn promotes the generation of testosterone from cholesterol. Then, testosterone signals the pituitary and hypothalamus to reduce their production of hormones through negative feedback regulation. Dihydrotestosterone (DHT), the active hormone, which is produced by the testis from testosterone. It is mostly (age related, but around 35%) linked to albumin and SHBG and delivered in blood to target tissues. Both cytoplasmic androgen receptors and nuclear are present in target organs [24].

Hoffman et al. [25] indicated through his study that suffices amount of protein enough for maintaining a positive nitrogen balance. Notably, the study also highlighted that increased protein intake did not influence resting hormonal concentrations, including testosterone levels, raising questions about the broader implications of protein supplementation on hormonal health and sexual function.

Research in humans shows mixed results about creatine affecting testosterone levels. The effect of creatine is via increased phosphocreatine stores. This may reduce overall metabolic stress, or the effect on testosterone secretion may be indirect through changes in the levels of sex steroid-binding globulin or the ratio of androgen to estrogen. Studies on rats show that creatine does not directly increase testosterone or other key reproductive hormones like Luteinizing Hormone (LH) or Follicle-Stimulating Hormone (FSH). Rahman et al. [26] pointed to strength and muscular mass are known to be enhanced by creatine. According to certain research, testosterone levels may rise in response to creatine intake, particularly when weight exercise is included. This may be the result of enhanced physical prowess and muscular growth, which might trigger the creation of testosterone. 

On the other side, Carbohydrates participated in increasing levels of testosterone in blood serum. Numerous research have examined the impact of carbohydrate supplementation on testosterone levels in male rats. In general, there are a number of ways that supplementing with carbohydrates might affect hormonal responses, including testosterone. First, Insulin levels can rise in response to carbohydrates, which may have an indirect effect on testosterone. Insulin may have an impact on testicular steroidogenesis and aid in the absorption of nutrients [27]. Second, sufficient consumption of carbohydrates may promotes general energy levels, which may be crucial for preserving hormonal equilibrium. The production of testosterone may decline when there is an energy imbalance [28]. This is supported by the results of craemeretal et al. [29], who referred that high intensive exercise with carbohydratic diet participate may stimulate the creation of greater testosterone, but in one condition of higher exercise intensity. Studies have indicated that testosterone levels can be increased particularly when accompanied with resistance training. However, the kind and quantity of carbohydrates taken can decline testosterone in the absence of exercise. 

In athletic settings, carbohydrate supplements can prevent energy deficits, which might otherwise impair reproductive functions. Energy deficits are known to reduce testosterone levels, sperm production and libido in male rats. Carbohydrate supplementation may modulate insulin levels, which in turn can influence the balance of other hormones, such as testosterone and Leuteinizing Hormone (LH). Insulin resistance has been linked to lower testosterone levels in both humans and animal models. Elevated blood glucose from excessive carbohydrate intake could potentially lead to oxidative stress, which may harm the testes and reduce sperm quality. Some studies suggest that excessive carbohydrate intake, particularly from refined sources, can increase oxidative stress in the body. This stress can damage the testes and negatively impact sperm motility and count. Antioxidants are sometimes supplemented to counteract these effects.

Carbohydrate consumption has been shown to influence testosterone levels by modulating cortisol. Cortisol, a stress hormone, competes with testosterone and elevated levels can suppress testosterone production. Carbohydrate supplements, particularly after exercise, might help reduce cortisol spikes and thus maintain healthier testosterone levels.

While the results histological part of the current study mentions the changes of epididymis with offering crucial information on recent testicular occurrences, epididymal histopathology ought to be conducted in conjunction with testicular histology. Testicular spermatozoa are unable to fertilize eggs. Spermatozoa develop their ability to fertilize throughout their passage through the epididymis. (such as capacitation, motility, response, acrosome and sperm-egg fusion capabilities). The main sources of accessory gland secretions are the Seminal Vesicles (SV), bulbourethral glands, prostates (often referred to as "Cowper's gland") and urethral glands [30].

The pathologist can be informed by the luminal contents in the epididymis of either the direct (primary, clear cells) or indirect (spermatids) toxicity. Epididymis Can be changed its function directly and in directly throughout different toxicants. There are two types of epidedymal damage, the first caused by androgen deprivation (from secondary spermatocytes to leydig cell toxicty in testis), the second damage is caused by a direct action of toxicants. Some of pathologist show that some of epidedymal functions may be altered without changing in tissue culture.

Further insight into the hormonal and reproductive consequences of protein supplementation is provided by the study conducted by Mutalip et al. [31], who investigated the histological changes in the testis of rats subjected to Anabolic Androgenic Steroids (AAS). Their findings revealed that AAS led to significant alterations in testicular structure, including reduced tubular size and disrupted spermatogenesis, which subsequently resulted in decreased testosterone secretion and sperm production. This study illuminated the potential long-term repercussions of hormonal disturbances induced by AAS, including diminished libido and testicular atrophy, suggesting that the intake of substances aimed at enhancing athletic performance may inadvertently compromise male reproductive health.

Animal studies have demonstrated that high-protein diets, particularly those supplemented with athletic protein powders, can have mixed effects on the sexual organs of male rats. Some studies indicate an increase in testicular size and function due to elevated testosterone, while others show signs of testicular atrophy or reduced sperm quality, potentially due to over-supplementation or imbalances in nutrient metabolism.

There is insufficient data to conclude that creatine directly controls spermatogenesis or sperm quality in male rats. Studies on oxidative stress in several organs have suggested that creatine may have some protective benefits on the male reproductive system. Adenosine triphosphate, or ATP, is transferred from the mitochondria to the contractile apparatus via the phosphocreatine shuttle, which is one of the several roles that creatine plays in spermatozoa. This process not only powers movement but also fertilization, cellular transport and other metabolic processes. Additionally essential to sperm activity, creatine kinase catalyzes the shuttle's energy replenishment. Adenosine triphosphate, or ATP, is transferred from the mitochondria to the contractile apparatus via the phosphocreatine shuttle, which is one of the several roles that creatine plays in spermatozoa. This process not only powers movement but also fertilization, cellular transport and other metabolic processes. Additionally essential to sperm activity, creatine kinase catalyzes the shuttle's energy replenishment [32].

In the current study, there is a sort of creatine toxity because of oxidative stress formation. oxidative stress can reduce sperm quality and function. Research focusing on rat sexual behavior (such as desire and mating activity) has not discovered any appreciable alterations as a result of creatine intake. Although the main effect of creatine is on physical performance, increased energy reserves may also theoretically improve the physical components of mating behavior.

Systemic Effects: Creatine might improve general health and systemic energy metabolism, potentially leading to improved overall vitality. This could indirectly benefit reproductive health by promoting better physical condition, although this is speculative and not confirmed through specific reproductive studies in male rats. The present study come to be disagree to the results of Allen [33]. Male, sexually inexperienced rats were given creatine supplements for seven weeks in order to determine the long-term effects on behavior and fertility. Male rats' ability to reproduce was not affected or enhanced by creatine supplementation, according to the findings of behavioral tests and anatomical examinations. Given that using creatine supplements doesn't affect the sexual system.

Seminal vesicles are secretory organs of the male genital tract found in certain eutherian species. The size, appearance and chemical makeup of the seminal vesicles vary according on the species androgens are necessary for the embryo's development, growth and secretory functions [34].

The seminal vesicle fluid contains very few proteins and those that do frequently aggregate into massive macromolecular clumps and become insoluble at certain pH levels, while it is not a prerequisite for conception, seminal vesicle fluid contributes to it in a number of ways. A biochemical model that demonstrates the possible importance of the interaction between the seminal vesicle protein unique to sperm and CAMP-dependent phosphorylation is provided [35]. 

It is still unknown how sperm cells connect molecularly with the proteins that secrete seminal vesicles. Still, a logical physiological picture is beginning to emerge. Rat and human epididymal sperm cell membranes contain an externally oriented CAMP-regulated protein kinase. The amount of externally orientated kinase enzyme is increased by the maturational epididymal transit." Some of the phosphorylated proteins that this enzyme use as substrates to absorb phosphates are proteins present in the seminal vesicle fluid, which subsequently adhere to the sperm cells [36].

The study of Ladyga et al. [37] refferred to D-aspartic acid role in the synthesis and release of sex hormones in humans have revealed that seminal plasma and spermatozoa contain D-Asp and that there is a direct correlation between D-Asp concentration and the quantity and motility of spermatozoa in semen. It was found that D-Asp may play an extraordinary role in spermatogenesis many athletes use dietary supplements, which are supposed to delay fatigue and improve the properties of the cardiovascular system. Most of these dietary supplements are based on carnitine, creatine and nitric oxide. At the same time, the issue of the effect of many dietary supplements on the sexual system of the male body and the mechanisms of their action on testicular cells remains controversial. In this regard, the purpose of this work was to study the effect of athletic supplements on the sexual organs of male rats and hormones. 

We can conclude through this study that athletic supplements (al-mass, Crea Pro and Isotope) are most familiar origins and used in gyms in Iraq. They are not pure components, but contain another gradients with lowest amount (like fats, vitamins, minerals.) depending on the company in addition to its basic components. These athletic supplements have negative effects on sex hormones and histology of reproductive organs in male rats without training. Many supplements effects are hormone-related, or they have an impact on hormone release. This is in turn effect sperm fertilizing ability. The relationship is complex and influenced by various physiological factors. External athletic supplements have negatice effect on the sexual ability to fertilize ova because of the histological changes in reproductive organs which is enough to cause male infertility. Their different effects are depending on the dosage, gradients, duration of intake, subject ability and training and the origion.

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