Bisphenol A is a chemical product as epoxy and polycarbonate resins that has a marked endocrine disrupting effect and environment due to its wide use in many fields and has an anti-androgen effect [28] and is prevalent in plastic used in food and beverage packaging such as bottles and water pipes [32]. Our work is the first work that revealed the improved effects of costus versus, the toxicity of Bisphenol A on the testis of the rat.
The present study showed that Bisphenol-A- reduced body weight and testes weight these reports parallel with reports by Samuel et al. [36] who postulate that treatment with BPA significantly reduction in the testes and body weights of the rats in comparison to the first group Anthonet and Orish [10]. He added that the cause of reduction in male albino Wistar rats may be due to reduced tubule size, a decrease in the number of germ cells, and degenerated spermatids after reproductive toxicity of lead. In addition to that Abdel-Halim et al. [2] showed that reduction may be due to a decrease in serum testosterone levels or may be due to decreased number of germ cells; suppression of spermatogonia cells also counts activity of the steroidogenic enzyme; these results were parallel with our results. Also, our present study showed that Costus had a remarkable ability to ameliorate the Bisphenol-A-induced decrease in body and testis weights, so Costus protects against the toxicity of Bisphenol A on the testis. This is consistent with reports by Khattab and Mansoury [30] who postulated that Costus afer leaf extract had a protective effect against testicular toxicity associated with cyclosporine, also our results were in coincidence with Abd El-Rahman et al. [1] who stated that administration S. lappa extracts significantly opposite TA-weight loss effects., they also added that this refinement may be due to improved function immunity and activities of antioxidant enzymes in rats. Improvement could be the consequence of the improvement of immune functions and antioxidant activities in rats.
Also, the present study, showed that Bisphenol A induced significant decreases of activity of antioxidant enzymes: (CAT), (GPx), (SOD) and (GSH), in the third group while significantly increased in the levels of Reactive Oxygen Species (ROS) as (MDA) and (NO) in tissues of testis of rat when compared with the first group. These results were parallel to the results of Samuel et al. [36]. Who stated that Bisphenol A caused a noticeable decrease of antioxidant enzymes with an increase in the levels of reactive oxygen species (ROS) they also added that these reductions of antioxidant enzymes may be due to enhanced utilization of GSH for detoxification of Bisphenol A induced free radicals or may be due to SOD converted the superoxide anion radicals into H2O2, which thereafter accumulated in the testis which inhibits CAT activity. Also, antioxidant enzymes can become inhibited with an elevation of lipid peroxidation as MDA Increase in reactive oxygen species (ROS) as MDA levels of the testis may cause a decrease in motility of sperm and destruction of the membrane of spermatozoa [24]. Apaydin et al. [12] added that enzymatic antioxidants (SOD, CAT, GPx, and GST) are useful in defending against the injury of cells and protecting cells and organs from the unfavorable effects of ROS by catalyzing the toxic H2O2 to water and oxygen. Also, these may cause the failure of the formation of sperm and steroid hormones, and hence, male sterility with infertility of males.
The present study deduced that Bisphenol-A caused toxicity on the male reproductive system in adult rats; these results were parallel to Kazemi et al. [29]. Who stated that Bisphenol A caused toxicity on the male reproductive system in adult rats due to elevation of ROS levels leads to oxidative stress, which also increases disorder caused by free radicals of oxygen and antioxidant enzymes of the cell [23].
Co-administration of costus in the present study is associated with a deficiency of MDA and H2O2 and a rise in the action of CAT, SOD, and GPx. These results were parallel with findings from Anthonet and Orish [10], who reports that these results were due to an inhibition of GPx1 gene code, and GPx4, associated with disturbance in the oxidation of protein; they also added that costus root extract showed obvious changes in oxidative stress of testis, hormonal, sperm analysis and histopathological changes caused by lead.
The present study showed that administration of Bisphenol A (third group) showed a significant reduction in the serum levels of testosterone, but no significant changes in FSH, PRL, and LH. as compared with the control and second groups, these findings are in coincidence with the results of Gonçalves et al. [20], who stated that the Bisphenol A is harm to TM3 cells of Leydig and damage their steroid action Also, our results were in coincidence with the results of Sohrab et al. [37], who reported that the decrease in the level of testosterone was most probably due to a deterioration in the activity of the Leydig and Sertoli cells; these results correspond with our ultrastructural results.
Kamel et al. [28] postulated that reduction of serum levels of testosterone is an indicator of chemical toxicity on the reproductive system,see also Samuel et al. [36] added that decreased levels of testosterone caused the failure of spermatogenesis and destruction of seminiferous epithelium, Leydig cell, and Sertoli cells, which is correlated with the histological and ultrastructural results of the present study.
Administration of costus root extract with Bisphenol A (fourth group) prevented the reduction of observed alterations in the serum levels of testosterone when compared with the third group; these findings were coincidence with the results of Anthonet and Orish [10], who postulated that costus Afer had a protective effect by elevating levels of plasma testosterone, in the treated groups to near normal.
4.1 Histopathology
The present work showed that the histological structure of control and second groups of rat testes are like normal testes of other mammals; these results were parallel to the results of Anthony [11].
The light microscope of the present study of the third group treated with Bisphenol A showed degeneration of seminiferous tubule, spermatogonia with vacuoles of sustentacular cells, degeneration in spermatocytes, spermatids, and degenerated interstitial cells, these were in correspondence with other studies reported by Samuel et al. [36]. Who reported that Bisphenol A caused degeneration of seminiferous tubules, and spermatocytes, also parallel with the results of Anthonet and Orish [10]. Who reported that Bisphenol A caused degeneration of spermatogonia with vacuoles of sustentacular cells [25]. Added that Bisphenol-A inhibits the growth of spermatogonia cells, developing Leydig cells, and steroidogenesis.
Co-administration of costus with Bisphenol A in the fourth group showed marked improvement in the testis of rats; these findings were parallel with the results of Domiaty et al. [17]. Who stated that the treatment with Costus extract and risperidone together showed an effective role in the improvement of damage caused improvement the pathological and anatomical changes they also added improvement of costus may be due to presence of flavonoids in the roots of Costus extract which has beneficial effects in certain diseases such as cancer and cardiovascular, neurological disorders.
Our results were parallel with the results of Anthonet and Orish [10]. Who reported that treatment with Costus afer showed marked improvement of histopathological changes induced by lead, they also added that extract of leaves of Costus had conservative effect versus damage of testis caused by lead.
Our data in Table 4 also showed that Bisphenol A caused a marked increase in the levels of the proapoptotic protein P53. Immunohistochemical staining has also revealed intense staining of P53; these results were in coincidence with the results of Yuan et al. [31]. Who reported that Bisphenol-A treatment caused an increase in p53 apoptotic cells in testicular tissues of the rat.
So, the rise of p53 in the present work detects the prospect of the occurrence of apoptosis after Bisphenol A. treatment.
Co-administration of costus with Bisphenol A in the fourth group caused marked improvement and decrease in the levels of proapoptotic proteins P53 these findings were parallel with the results of Gules et al. [21]. Who stated that costus extract contains flavonoids that improve the reduction of oxidative stress, which has emerged as a cell protective agent when exposed to activity scavenging of free radicals that cause damage to cell structures, leading to inhibiting DNA damage. So costus appeared to have anti-apoptotic properties.
4.2 Ultrastructural study
The ultrastructural examination of the present study showed that it showed enlarged vacuoles, swollen mitochondria, cell debris, and cytoplasmic vacuoles of Sertoli cells; these findings were parallel with the results of Gurmeet et al. [22]. The current study showed swollen degeneration of spermatogonia cells, primary spermatocytes with excessive lipid droplets, irregular nuclear membrane, and pyknotic nuclei; these results were parallel with the findings of Toyama et al. [39]. Who stated that Bisphenol A caused several degenerative effects on spermatogonia like the disappearance of the nuclear envelope and numerous apoptotic changes. Our ultrastructural results also revealed Bisphenol A caused damages of spermatogonia cells and spermatocytes as condensation of chromatin material, the disappearance of the nuclear envelope, and numerous apoptotic changes; these results were parallel with the findings of Tushara et al. [40]. Who postulated that Bisphenol-A caused ultrastructural several degenerative effects in the form of the presence of vacuoles in mitochondria of spermatids, Sertoli, Leydig, and peritubular myoid cells as compared to control.
In addition to that Zaki et al. [42]. Reported that an increase of vacuoles in the cytoplasm of the spermatogonia and sustentacular cells might be caused by swelling of the smooth endoplasmic reticulum that demonstrates changes in the permeability of cells in addition to that the ultrastructural changes may be due to bisphenol A had apoptosis in various spermatogonia and destruction of basement membrane which has the main function in the integrity of testicular tissues.
The testis of rats of the fourth group Bisphenol A and costus-treated group, showed marked improvement in spermatogonia. Normal spermatids, also primary spermatocytes and sperms appeared nearly normal as condensation of chromatin material. These results were parallel with the findings of Domiaty et al. [17]. Who postulated that ultrastructural changes of testis of treated rats with risperidone and costus fourth group identified that developmental stages of spermatogenic epithelium including spermatogonia, normal spermatids, primary spermatocytes, and sperms were detected also Sertoli cells appeared as elongated cells with its giant nuclei, prominent nucleoli, ER and mitochondria. The intertubular space was full of a moderate number of Leydig cells; they also added that treatment with Costus inhibited the histopathological alterations induced by risperidone within the testis.