Number of cells in Peyer’s patches in the germinal center region of the groups Control - TC, Bisphenol A - BPA, Bisphenol A + Selenium - BPA+Se and Selenium - SE, (p<0.05) (* versus groups CT; BPA+Se and Se).
4. DISCUSSION
This study showed that BPA treatment promoted an alteration in the epithelial layer of PP´s, inducing possible apoptotic processes and morphological changes in goblet cells. These results were also observed in the work of Feng et al. (2019), where they found that in addition to leading to apoptotic processes, BPA also reduced the number of goblet cells, inhibited the expression of tight junction proteins, and altered the diversity and structural composition of the microbiota, eventually reducing intestinal permeability in mice.
These results do not corroborate the findings of Özaydin et al. (2018), where no changes were observed in their experimental groups. The differences in results are due to the time of exposure to the substance and the dose administered in the groups studied.
Furthermore, publications in Romania reveal that the toxicity caused by BPA has been able to cause tissue changes in other organs. The effects of BPA depend on several molecular and epigenetic mechanisms that determine whether the endocrine or reproductive system is affected, whereby pre-existing benign lesions can become cancerous (Dumitrascu, et al., 2020).
Ambreen et al. (2019) performed an in vivo evaluation in BPA-intoxicated rats and demonstrated similarities with the present study, in which histological analysis revealed changes in the intestinal tissue of BPA-treated rat groups compared to the control. In addition, the group treated with low doses of BPA had a lesion in the small intestine. On the other hand, the group that received high doses had a significant fissure in the intestine, with hyperplasia of the cells in the lamina propria along with shrinkage and rupture of the villi. Therefore, the severity of the trauma depends on both the time of exposure and the amount of dose received.
The protective capacity of Se has also been documented against BPA-induced oxidative stress. Rafiee et al. (2021) found that, overall, Se reduced mitochondrial oxidative stress and effectively improved mouse sperm survival and motility, suggesting that Se may improve mitochondrial damage caused by BPA and impaired mouse sperm quality by preventing oxidative stress.
Khalaf et al. (2018) concluded that co-administration of Se and nano-Se (NSe) attenuated BPA-induced reproductive toxicity via improved antioxidant activity, genetic alterations, and restoration of testicular tissue almost like their control group. These results indicated that both forms of Se and NSe can be used as reproductive protective agents against the detrimental effect induced by BPA. Our study corroborates these studies, where it is possible to perceive a possible protective effect of the Se against the injury induced by exposure to BPA. In addition, we noticed a reduction in inflammation caused by the administration of BPA in the follicular region of the Peyer’s patches.
Our results indicate that there was a reduction in the process of cell proliferation in the epithelial layer of Peyer’s patches in the BPA group. Previous studies showed that BPA was able to inhibit cell proliferation in human colonic epithelial and goblet cells (QU et al., 2018; ZHAO et al., 2019). These findings indicate BPA’s potential to disrupt intestinal barrier function. However, our study did not correlate the effects of BPA and the induction of apoptosis due to the absence of markers in PP´s. Qu et al. (2018) demonstrated that this substance promoted cell apoptosis, weakening the functions of the intestinal epithelial barrier, altering intestinal permeability.
Our results also corroborate the work of Wang et al. (2021) where they concluded that BPA exerts deleterious effects on the epithelial layer, in which it activates an innate immune response that immediately disrupts the balance between cell damage and repair. Consequently, the intestinal epithelial barrier and permeability are disrupted, leading to intestinal damage.
The data from this study showed that Se has a protective effect against epithelial barrier dysfunction caused by BPA. This study shows that Ki-67 expression levels were increased in animals given BPA. The protective effect of Se under conditions of oxidative stress has already been reported by Dou et al. (2023), where they demonstrated that the protective effect of SeNPs on intestinal epithelial barrier injury is closely associated with the mitochondria-lysosome crosstalk signaling pathway mediated by TBC1D15/Rab7.
Canter et al. (2021) pointed out that selenium had a potential role in the integrity of the intestinal barrier and structural changes in glandular goblet and mucin-producing cells in the mucosa and submucosa of the colon. Liu et al. (2020) showed that Se attenuated oxidative stress-induced intestinal mucosal disruption, which was associated with elevated mucosal antioxidant capacity and improved intestinal barrier functions. Xu et al. (2018) also reported that selenium particles promoted the growth and proliferation of porcine intestinal epithelial cells, human colonic epithelial cells, and macrophages derived from human acute monocytic leukemia cells.
These results corroborate the findings of Afzal et al. (2022) where in their study a significant reduction in hemoglobin (Hb), lymphocytes, globular volume (PCV), red blood cells (RBC) and monocytes was observed in a dose-dependent manner compared to their control group. They suggest that due to the toxic accumulation of BPA, there was a decrease in pH, plasma volume in the blood, as well as a low oxygen supply to the red blood cells. Tran et al. (2020) found that BPA had potential negative effects on the T cell response as it decreased telomerase activity via an ER/GPR30-ERK signaling pathway.
Our findings do not corroborate those of Ye et al. (2023) who found an increase in immune cells infiltrated in the colon and increased expression of GHSR (hormone receptor secretagogue) and pro-inflammatory cytokines and chemokines, such as Il6e Ccl2, in the colon mucosa. However, our results are similar to those of this study, since exposure to BPA increased the number of macrophages in the groups exposed to BPA. The authors suggested that GHSR signaling from the nutrient-sensitive ghrelin receptor is involved in modulating the effect of BPA on macrophages and when GHSR expression increases, it activates innate immunity systemically.
Özaydeun et al. (2018) noticed that CD8+ and CD4+ lymphocytes in ileal PP´Ss, exposed to BPA, were mainly located in the interfollicular region (IFR). Notwithstanding, there was a significant increase in CD8+ lymphocyte count and a decrease in CD4+ lymphocytes compared to the control and vehicle groups. It is worth noting that in physiological situations there is a predominance of CD4+ in PP´s. Our study does not corroborate these findings, suggesting that this change in the number of leukocytes may be related to the time of exposure and BPA dosage, significantly altering the immune response performed by PP´s.
We observed that in the animals supplemented with Se, the defense cells remained in a similar amount to the control group. The literature shows that selenium can act as an antioxidant, protecting against oxidative stress, aiding in cell survival and growth, thus playing a chemopreventive role (SELENIUS, et al., 2010; RAZAGHI, et al., 2021).
Köse; Naziroğlu (2014) showed in their work that Se supplementation also protected neutrophils from endogenous oxidative stress. Huang; Rose; Hoffmann (2012) suggest that selenium supplementation, for the most part, is immunostimulating, which is measured by a wide range of parameters, including T cell proliferation, NK cell activity, innate immune cell functions, and many others depending on the dose.
Our study corroborates these findings, as intense cell proliferation was observed in PP´s and we suggest that Se is necessary for optimal immune function, in addition to minimizing oxidative damage caused by BPA. However, selenium at the administered dose did not show a significant difference in the number of defense cells when compared to the control group, suggesting the need for a more refined evaluation of how selenium affects different types of immune responses.
5. CONCLUSIONS
According to the results, it was observed that the ingestion of Bisphenol A above the levels considered safe resulted in changes in the tissues of Peyer’s patches, causing a reduction in cell proliferation. In addition, BPA decreased the population of immune cells in these structures. Crucially, selenium has been shown to have a possible protective effect against BPA-induced toxicity. The results suggest that Se plays a crucial role in the immune functions of Peyer’s patches. The findings of this study not only contribute to the current understanding of the immune function of these structures, but also provide a solid foundation for future research. It is believed that this work can broaden our understanding of the impacts of BPA and serve as a starting point for further investigation.
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