| BackgroundWith the rapid development of communication technology,human beings are exposed to a large amount of electromagnetic radiation every day,which has caused a great impact on the health of the body.At the same time,infertility has become a widespread concern in today’s society,and infertility caused by male factors cannot be ignored.Previous studies have shown that decreases in male fertility are associated with exposure to electromagnetic radiation.Therefore,it is urgent to explore safe and effective electromagnetic radiation protection agents.In recent years,molecular hydrogen has gradually become a hot topic in the research field of biological injury protection,because it can exert many effects,such as anti-oxidation,anti-apoptosis,anti-inflammation and anti-radiation.Therefore,whether molecular hydrogen can exert its protective effect on electromagnetic radiation damage of male reproductive system and become a potential electromagnetic radiation protection agent has attracted our research interest.To this end,we conducted the following studies.First,the damage effect of NEMP exposure on the male reproductive system was clarified and the electromagnetic radiation damage model for the male reproductive system was established.Then molecular hydrogen intervention was conducted to clarify whether the damage to the male reproductive system after NEMP exposure was caused by protective effects of molecular hydrogen,and what specific protective effects were produced.Finally,the possible protective mechanism of molecular hydrogen was explored.Contents1.Effects of NEMP exposure on the reproductive system,spermatogonia and Leydig cells in male rats.2.Effects of molecular hydrogen on reproductive system,spermatogonia and interstitial cells of testis in male rats exposed to NEMP.3.Effects of molecular hydrogen on ROS,mitochondrial membrane potential and mitochondrial apoptosis pathway in spermatogonia and Leydig cells after NEMP exposure.4.Effects of molecular hydrogen on antioxidant enzymes,oxides and antioxidants in testis of rats exposed to NEMP.5.Analysis of transcriptomics and metabolomics of rat testicular tissue.Methods1.Effects of NEMP exposure on male reproductive system.The testicular tissue of male rats exposed to NEMP was analyzed by HE staining,Tunel staining,Western Blot and Brd U staining.The serum testosterone level of male rats was measured by ELISA to analyze the effects of NEMP exposure on the reproductive and endocrine system of male rats.The sperm parameters of rat cauda epididymis were detected by CASA,and the effects of NEMP exposure on the sperm of male rats were analyzed.The effects of NEMP exposure on the viability of spermatogonia and Leydig cells were analyzed by CCK8 and Ed U staining.The effects of NEMP exposure on the apoptosis of spermatogonia and Leydig cells were analyzed by Annexin V/PI double-staining flow cytometry and Western Blot.2.Protective effects of molecular hydrogen on NEMP exposure injury in male reproductive system.The protective effects of molecular hydrogen on testicular tissue of male rats after NEMP exposure were analyzed by HE staining,Tunel staining,Western Blot and Brd U staining.The serum testosterone level of male rats was detected by ELISA,and the protective effect of molecular hydrogen on reproductive endocrine system of male rats after NEMP exposure was analyzed.To analyze the protective effect of molecular hydrogen on sperm of NEMP-exposed male rats,the parameters of rat epididymal sperm were detected by CASA.The protective effects of molecular hydrogen on spermatogonia and Leydig cells after NEMP exposure were analyzed by CCK8,Ed U staining,Annexin V/PI double-staining flow cytometry and Western Blot.3.Protective mechanism of molecular hydrogen on NEMP exposure injury of male reproductive system.The levels of intracellular ROS,mitochondrial membrane potential and the release of cytochrome C in mitochondria were detected to clarify the relationship between molecular hydrogen and intracellular ROS,mitochondrial membrane potential and mitochondrial apoptosis pathway.The T-AOC,SOD,GSH,GSH-Px,LDH,CAT,SDH and MDA contents in the testicular tissue of rats were detected to identify the effects of molecular hydrogen on the antioxidant enzyme activities,GSH and MDA contents in the testicular tissue of rats after NEMP exposure.Through the analysis of transcriptome,metabolomics and the combination of transcriptome and metabolomics of rat testis,we explored the target molecules and target pathways of molecular hydrogen in the protection of male reproductive system NEMP exposure injury.Results1.Effects of NEMP exposure on male reproductive system.1.NEMP exposure resulted in pathological changes in rat testis,but had no significant effects on testicular organ index and diameter of seminiferous tubules.2.NEMP exposure induced apoptosis in rat testicular tissue.3.NEMP exposure resulted in the decrease of cell viability in rat testis.4.NEMP exposure decreased the serum testosterone concentration of rats.5.NEMP exposure decreased sperm viability and motility in the cauda epididymis of rats,but had no significant effect on sperm concentration,deformity rate and motility parameters.6.NEMP exposure decreased the viability of spermatogonia and Leydig cells in mice.7.NEMP exposure induced apoptosis in spermatogonia and Leydig cells.2.Protective effects of molecular hydrogen on NEMP exposure injury in male reproductive system.1.Molecular hydrogen could alleviate the pathological changes of rat testis caused by NEMP exposure.2.Molecular hydrogen could alleviate the apoptosis of rat testis caused by NEMP exposure.3.Molecular hydrogen could alleviate the cell viability decrease of rat testis caused by NEMP exposure.4.Molecular hydrogen could alleviate the decrease of serum testosterone concentration in rats caused by NEMP exposure.5.Molecular hydrogen could alleviate the decrease of sperm motility caused by NEMP exposure but has no significant effect on sperm survival rate.6.Molecular hydrogen could alleviate the decrease in the viability of spermatogonia and Leydig cells caused by NEMP exposure.7.Molecular hydrogen could alleviate the apoptosis of spermatogonia and Leydig cells caused by NEMP exposure.3.Protective mechanism of molecular hydrogen on NEMP exposure injury of male reproductive system.1.Molecular hydrogen could decrease the intracellular ROS level after NEMP exposure.2.Molecular hydrogen could alleviate the decrease of cell mitochondrial membrane potential after NEMP exposure.3.Molecular hydrogen could inhibit the release of cytochrome C from mitochondria after NEMP exposure.4.Molecular hydrogen could increase the activities of GSH-Px and LDH in the testis of rats after NEMP exposure and accelerate the recovery of SOD activity and MDA concentration.5.Molecular hydrogen treatment alone could up-regulate the expression of 106 genes,down-regulate the expression of 113 genes,decrease the level of 16 metabolites and increase the level of one metabolite in the testicular tissue of rats.6.NEMP exposure resulted in up-regulation of 77 genes and down-regulation of 128 genes,decreased levels of 12 metabolites and increased levels of 3 metabolites in rat testis.7.Molecular hydrogen intervention after NEMP exposure resulted in up-regulate the expression of 97 genes,down-regulate the expression of 102 genes,increase the levels of 16 metabolites and reduce the levels of 2 metabolites in the testicular tissue of rats.ConclusionNEMP exposure resulted in in testicular tissue damage,decreased serum testosterone concentration,decreased sperm viability and motility in rats,and damage to spermatogonia and Leydig cells.Molecular hydrogen could alleviate the damage to testicular tissue,the decrease of serum testosterone concentration,the decrease of sperm motility,and the damage to spermatogonia and Leydig cells caused by NEMP exposure.Molecular hydrogen plays a protective role against NEMP exposure,which is related to its efficacy in reducing ROS production,protecting mitochondrial membrane potential and inhibiting mitochondrial apoptotic pathways,as well as enhancing antioxidant enzyme activity.The combined transcriptome-metabolomics analysis revealed that molecular hydrogen played a protective role against NEMP exposure and was related to the impact on metabolic pathways. |
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