Effect And Mechanism Of Restraint Stress On Reproductive Function And The Investigation Of Anti-stress In Male Mouse

In the current situation of social life, all negative emotions constitute the variety of life stress events, including the fast-paced life, pressure from career and family, as well as congestion, ambient noise, and so on. Male infertility is characterized as asthenozoospermia, azoospermatism, oligospermia. The possible consequence is declination of humane reproductive capacity, therefore male infertility caused by stress is urgent to be solved. This experiment was conducted to study the effect of stress-induced on adolescent and adult male mouse testis morphology pathology, germ cell apoptosis, testosterone synthesis, and spermatogenesis using a mouse model of restraint stress. We also detected the expression of endoplasmic reticulum stress pathway proteins in mouse testis. We tried to illuminate the mechanism of testicular germ cell apoptosis, spermatogenesis dysfunction under restraint from the aspect of endoplasmic reticulum stress. Furthermore, we investigated the effect of 4-PBA, GHRL and CRY on the ER stress, oxidative stress and inflammatory response, respectively. 1 The effect of restraint stress on reproductive function in male puberty miceObjective: The current study was conducted to examine the effect of chronic stress on the testicular function and spermatogenesis in the adolescent male mice. Results: The sperm deformity rate was significantly increased after stress. The weight of testis and epididymis were significantly reduced in stress group and recovery group, but the relative weight of testis and epididymis was significantly increased. Stress significantly reduced testosterone level in serum, but it continued to decrease after 24 h recovery. Stress significantly increased the rate of apoptotic germ cells, but restored after 24 h recovery. The m RNA expression levels of St AR, 3β-HSD and P450 scc were significantly inhibited in stress and recovery group compared with control group. Conclusion: The stress on adolescent mice spermatogenesis and testosterone have a significant role in the synthesis of damage, stress damaging effects of puberty in mice spermatogenesis and testosterone synthesis of testosterone may be associated with testicular St AR and reduce synthase expression and apoptosis of germ cells. 2 The mechanism of chronic stress-induced on reproductive development in maleadult miceObjective: The current study was conducted to examine the effect of chronic stress on the testicular function and spermatogenesis in the adult male mice. Results: The sperm deformity rate was significantly increased after stress. The weight of testis and epididymis were significantly reduced in stress group and recovery group, but the relative weight of testis and epididymis was significantly increased. Stress significantly reduced testosterone level in serum, but it continued to decrease after 24 h recovery. Stress significantly increased the rate of apoptotic germ cells, but restored after 24 h recovery. The expression levels of St AR gene and related 3β-HSD and P450 scc were significantly inhibited after stress in stress and recovery group. Conclusion: The stress on adolescent mice spermatogenesis and testosterone has a significant role in the synthesis of damage, stress damaging effects of puberty in mice spermatogenesis and testosterone synthesis of testosterone may be associated with testicular St AR and reduce synthase expression and apoptosis of germ cells. 3 The effect of 4-PBA on restraint stress-induced ER stress in male adult miceObjective: The current study was conducted to examine the mechanism and the effect of chronic stress on the testicular function and spermatogenesis in the adult male mice. Results: The sperm deformity rate was significantly increased after stress, but PBA treatment inhibited the increase. The weight of testis and epididymis were significantly reduced in stress group in comparsion with control group, but PBA inhibited the decrease. Stress significantly reduced testosterone level in serum, but it reversed pretreated with PBA. Stress also significantly increased the rate of apoptotic germ cells, but restored after PBA treatment. The expression levels of St AR gene and related 3β-HSD and P450 scc were significantly inhibited after stress, but the inhibition was alleviated in PBA+stress group. Furthermore, stress significantly up-regualted the Bax and down-regulated Cytc and Bcl-2 genes in mitochondria. Inversely, the expression of Cytc and Bax were up-regulated in cytosol. Conclusion: Stress has a significantly harmful effect on spermatogenesis and testosterone synthesis in adult mice, which may exist some relationship with germ cell apoptosis. ERS signal pathway mediated stress-induced germ cell apoptosis. 4 The effect of GHRL on restraint stress-induced ER stress in male adult miceObject: The research is to investigate the effects and mechanism of GHRL on ERS of mice testis induced by restraint stress. Results: Compared to other groups, the weight of body, testis and epididymis of stress group has a significant reduction. On the contrary, the index of testis and epididymis both significantly raised. In group, the result is that the weight of body, testis, epididymis and the level of the testosterone both are significantly higher than those in stress group. The seminiferous epithelium of stress group becomes thin and the structure of seminiferous tubule is disorganized and irregular. The spermatogenic cells in all levels reduced and became disordered. The count of mature sperm in Seminiferous tubule decreased and the deciduous spermatogenic cells could be seen at the same time. In GHRL group, the testicular tissue was normal and the cells in Seminiferous tubule had an organized arrangement with normal leydig cells among them. The apoptosis of germ cells in stress group is significantly severe and the contrary situation happened in GHRL group. The decline of the expression of STAR, P450 Scc and 3β-HSD deduced by ERS could be inhibited by GHRL. Also, GHRL could inhibited the m RNA and the protein expression of XBP-1, p-JNK, CHOP, e IF2α, GRP 78 and ATF6. After the injection of GHRL, the m RNA expression of XBP-1, p-JNK, CHOP, e IF2α, GRP 78 and ATF6 deduced by ERS could also be inhibited. Conclusion: GHRL inhibits the expression of some protein in endoplasmic reticulum and the apoptosis of germ cells in testis deduced by stress which may be through the inhibition of ERS. 5 Effects of β-cryptoxanthin Against Cadmium-induced Oxidative Stress in the RatTestisβ-cryptoxanthin(CRY), a major carotenoid of potential interest for health, is obtained naturally from orange vegetables and fruits. The pathological and TUNEL findings revealed that CRY ameliorated the Cd-induced testicular histological changes and germ cell apoptosis in the rats. Furthermore, the Cd-induced decrease in the testicular testosterone(T) level was attenuated after CRY administration(p<0.05). The administration of CRY significantly reversed the Cd-induced increases in the lipid peroxide(LPO) and malondialdehyde(MDA) levels(p<0.01). The testicular antioxidants superoxide dismutase(SOD), catalase(CAT) and glutathione(GSH) were decreased by treatment with Cd alone, but were restored by CRY co-treatment. These results demonstrated that the application of CRY can enhance the tolerance of rats to Cd-induced oxidative damage and suggest that it has promise as a pharmacological agent to protect against Cd-induced testicular toxicity. 6 Anti-inflammatory potential of β-cryptoxanthin against LPS-induced inflammationin mouse Sertoli cellsβ-Cryptoxanthin(CRY), a major carotenoid pigment, can inhibit inflammatory gene expression in mice with nonalcoholic steatohepatitis. In the present study, we examined the anti-inflammatory effects of CRY on lipopolysaccharide(LPS)-induced inflammation in mouse primary Sertoli cells and the possible molecular mechanisms behind its effects. The results showed that CRY significantly inhibited LPS-induced decreases in cell viability and in the percentage of apoptotic cells. Moreover, CRY inhibited the LPS-induced up-regulation of tumor necrosis factor α(TNF-α), interleukin-10(IL-10), interleukin-6(IL-6) and interleukin-1β(IL-1β) in Sertoli cells. In addition, CRY significantly limited the LPS-induced down-regulation of AR, HSF2, CREB, FSHR, INHβB and ABP in Sertoli cells. Western blotting analysis showed that CRY significantly suppressed NF-κB(p65) activation as well as MAPK phosphorylation. All the results suggested that CRY suppressed inflammation, possibly associated with the NF-κB activation and MAPK of phosphorylation. Thus, CRY may possess therapeutic potential against inflammation-related diseases.To sum up, the conclusion of this study(1) stress has significant damage on spermatogensis and testosterone synthesis, and induces germ celluar apoptosis on adolescent and adult mice;(2) endoplasmic reticulum stress signaling pathway is mediated stress-induced testicular germ cell apoptosis;(3) 4-PBA inhibits restiant stress-induced ER stress and germ cell apoptosis;(4) GHRL inhibits endoplasmic reticulum associated protein and testicular germ cell apoptosis induced by stress;(5) application of CRY can enhance the tolerance of rats to Cd-induced oxidative damage;(6) CRY suppressed inflammation, possibly associated with the NF-κB activation and MAPK of phosphorylation.