Preliminary Study On The Effect And Mechanism Of Infertile Male Psychology And Dehydroepiandrosterone On Semen Quality In A Rat Model Of Oligoasthenozoospermia Complicated With Depression

PART1 PSYCHOLOGY OF INFERTILE MEN UNDERGOING ART AND EFFECT OF ANXIETY/DEPRESSION ON THE SEMEN QUALITY AND SERUM HORMONE[Objective]:To study the psychological status of infertile male in the first time of Assisted Reproduction Technique and its effect on semen quality and hormones,[Methods]:Through clinic questionnaires,SCL-90,HADS and SSRS rating scale,study analyzed the psychological status of 1173 infertile male undergoing first ART in cross-section,assess relative risk factors and then divided them into high-score group(?9 points)and low-score group(<9 points)according to the HADS scale scores,evaluated the semen quality of patients with different groups and the level of serum sex hormones(FSH,LH,PRL,and Testosterone).[Results]:Comparing with national norm,infertile male undergoing the first time of ART possessed increased scores of hostility of SCL-90,HADS(P<0.05)and there was no statistically significant difference in the rest of SCL-90 factors,the sperm concentration of HADS high score group?9(points)decreased obviously than that of HADS low score group(<9 points)(P<0.05),Meanwhile,the level of serum testosterone dropped while that of prolactin rose,and the difference was statistically significant(P<0.05).[Conclusion]:1 Comparing with nation norm/normal population,infertile men undergoing first ART treatment and national norm,hostility scores of SCL-90,scores of HADS increased.There was no statistically significant difference in the rest of SCL-90 factors.2 HADS scores of infertile men undergoing first ART treatment are higher than normal people,and HADS high score group?9(points)had significantly decreased sperm concentration and level of serum testosterone.Part 2 Experimental study on the treatment of DHEA on the semen quality of the animal model[Objective]To build the oligoasthenospermia/Depression animal models of rats and evaluate the effect and mechanism of DHEA improved on semen quality of the model.[Methods]We applicated intraperitoneal injection of cyclophosphamide to build rat model with oligoasthenozoospermia,whose semen quality decreased obviously compared with normal group;And we applicated CUMS to build rat model with depression,which showed significant differences in Sugar preference test,Forced-swimming test and Open-field test compared with the control group:model group fewer Sugar preference,significantly more motionless time in forced swimming test,significantly more rest time in Open-field test and diminished obviously exploring ability for the model group,indicating that quality of model was good.We divided 84 model rats with oligoasthenozoospermia/deprssion into two experimental groups:one group of 36 to expriments of Open-field test,Forced swimming,another group of 36 to expriments of sucrose preference test.The group of 36 to expriments of sucrose preference test divided into four groups:DHEA group,fluoxetine group,model rat with oligoasthenozoospermia and depression and control group(rats with oligoasthenozoospermia).We evaluated threatpeutic effect of DHEA and fluoxetine on rat models.[Results]The semen concentration of rat model(9.64±2.94)decreased obviously compared to this of the normal control group(16.88±3.28).The rat model showed statistical significant difference:sucrose preference(0.34 ±0.03 vs 0.72±0.11),Open-field test(257.43±52.87 vs 186.14±42.40),the immobility time(117.1±32.67 vs 54.94±12.19),We treated rat model group with DHEA,fluoxetine respectively,results showed that comparing with untreated model group,sperm concentration increased of DHEA group,The study indicated that DHEA is beneficial for increasing sperm concentration.[Conclusion]1.Using the cyclophosphamide and CUMS method to establish the rat oligoasthenozoospermia/depression model was relatively simple and well.2.The treatment of DHEA and fluoxetine increased the semen concentration of rat model.Part 3 Experimental study on the treatment of DHEA on testosterone levels and oxidative stress in the rat model of oligoasthenozoospermia/depression[Objective]study to effect of DHEA on the levels of serum testosterone,prolactin and oxidative stress of model rat testicular tissue.[Methods]The sex hormone levels and oxidative stress of model rat were detected through Sex hormone ELISA kit,MDA and GSH test kit.[Resluts]MDA(Oxidative stress index)was significantly increased(M vs S)(5.46±0.58 vs 3.31±0.31)(P<0.05),while GSH(the antioxidant stress index)decreased further(M vs S)(0.13±0.06 vs 0.16±0.08),but no statistical significance.Serum total testosterone(Mvs S)(3.36±1.29 vs 1.35±0.31)and free testosterone(Mvs S)(0.11±0.05 vs 0.05±0.01)decreased(P<0.01),and serum prolactin(Mvs S)(8.50±0.30 vs 7.93±0.38)levels were increased.And after treatment with DHEA model animal serum testosterone(2.68±0.55)and free testosterone levels(0.09±0.02)increased,MDA decreased and GSH increased,improving the oxidative stress.fluoexitine also improved too.[Conclusion]Depression could decresed to tissue GSH level and increased to tissue MDA level.DHEA,fluoexitine improved the level of GSH of testis tissue,and decreased level of MDA.Part 4 Effect of DHEA on the expression of CREB,Nrf2 and HO-1 in testicular tissue of the rat model of oligoasthenozoospermia/depression[Objective]To explore the possible mechanism of DHEA using relativelysignal pathway of Nrf2,HO-1,CREB protein to improve the quality of semen in the rat model,and provide the experimental base of clinic practice.[Methods]We analyzed the effect of DHEA,fluoxetine on the expression of protein Nrf2,HO-1 and CREB in the testicular tissue of model rats with oligoasthenozoospermia/depression,Using methods such as Western blot,RT-PCR and immunohistochemistry.[Results]Differing from their low expression in testicular tissue of normal rats,protein of Nrf2,HO-1 and CREB were distinctly up-regulated expression in rat model with oligoasthenozoospermia/depression.In immunohistochemistry,protein CREB,Nrf2,HO-1 which mainly distributed in the cytoplasm and nucleus were up-regulated expression in the leydig cells and spermatogenic cells in seminiferous tubules of rat model group.In RT-PCR,compared with control group,mRNA in rat model group of CREB,Nrf2,HO-1 increased(P<0.05).In Western blot,protein in rat model group the expression rate of CREB,Nrf2,HO-1 increased related to control group,and the difference was statistically significant(P<0.05).Application of DHEA,fluoxetine can promote protein CREB,Nrf2 and HO-1 expression to different degrees,but the expressiom will decrease after testicular tissue repairing step by step.[Conclusion]1 Application of DHEA,fluoxetine can promote protein CREB,Nrf2 and HO-1 expression to different degrees,but the expressiom will decrease after testicular tissue repairing step by step.2 Both DHEA and fluoxetine promoted the sperm concentration,through improving the expression of protein CREB,Nrf2,HO-1 and oxidative stress resistance.DHEA also played a preferable role in the ascension of serum testosterone level than fluoxetine.