The Effects Of Emodin On The Apoptosis Of Spermatogenic Cell Induced By Varicocele

Varicocele is a common andrology disease defined as abnormal dilatation and tortuosity of veins in the pampiniform plexus of the spermatic cord,which leads to spermatogenesis dysfunction and male infertility.The incidence of varicocele in adolescents is around 15%and varicocele accounts for 35-40%in infertility patients.Thus varicocele is the most common disease which leads to infertility.Emodin is an effective component of traditional Chinese medicine rhubarb,which has anti-tumor,anti-inflammation,anti-infection,anti-oxidative stress,immunosuppression and other effects.It has clinical application value in the treatment of pancreatitis,glomerulonephritis,hepatitis,asthma,atherosclerosis and various tumors.However,the research on emodin in the treatment of varicocele is very limited.Our study is divided into two parts,the rat model and the cell model were established to investigate the efficacy of emodin protecting seminiferous epithelial cell and the mechanism of that.Our study may provides a new idea for the treatment of male infertility caused by varicocele.Part?The effects of emodin on the apoptosis of spermatogenic cell in rat testes induced by varicoceleObjectives:to investigate the effects of emodin on the apoptosis of spermatogenic cell in rat testes induced by varicocele.Methods:40 male Sprague Dawley rats were randomly distributed into 4 groups:control group(group A),control+emodin group(group B),varicocele group(group C)and varicocele+emodin group(group D).The number of rats in each group is 10.Group A and group B were sham surgery groups.The rats in group A and group B were only opened the abdominal cavity but didn't receive other surgery intervention.Group C and group D were surgery groups and the rats in group C and group D were used to establish varicocele models.The rats in group B and group D received intraperitoneal injection of emodin 20mg/(kg·d)after undergoing surgery.On the other hand,the rats in group B and group D received intraperitoneal injection of normal saline as equal volume.60 days after surgery,left orchiectomy were operated in all groups.HE stainings were applied to observe the changes of morphology and structure of seminiferous tubules.TUNEL stainings were applied to observe the apoptotic seminiferous epithelial cells and the rates of apoptosis were calculated.The expression of Bax and Caspase-3 were detected by western blot and RT-q PCR.The oxidative stress level of testicular tissue was evaluated by measuring MDA and SOD.Results:HE staining showed the structure of the seminiferous tubules became disorganized and the number of seminiferous epithelial cells decreased in group C compared with group A.But the degree of morphologic destruction in group D was milder in contrast with group C.TUNEL staining showed the number of apoptotic cells in group A and group B were few.However,the number of apoptotic cells in group C significantly increased.And the number of apoptotic seminiferous epithelial cells in group D were less than that in group C.In experiment of western blot and RT-q PCR,The level of expression of Bax and Caspase-3 in group C were significantly higher than the level in group A.And expression of Bax and Caspase-3in group D decreased compared with group C.The MDA level of group C was higher than that of group A and SOD expression was lower than that of group A.Compared with group C,the MDA level of group D was lower and SOD expression was higher.Conclusions:Varicocele leads to structure destruction of seminiferous tubules,decrease in the number of seminiferous epithelial cells and increase in the number of apoptotic seminiferous epithelial cells,the expression of Bax and Caspase-3 and the level of oxidative stress.Emodin treatment to rats with varicocele inhibits seminiferous epithelial cell apoptosis,lowers the expression of Bax and Caspase-3and the level of oxidative stress,and alleviates the damage to seminiferous tubules and seminiferous epithelial cells.Part?The effect of emodin on the apoptosis of spermatogonia caused by hypoxiaObjectives:to investigate the effect of emodin on the apoptosis of spermatogonia caused by hypoxia.Methods:GC-1 cells were selected and four groups were set up as control group(group A),control+emodin group(group B),model group(group C),model+emodin group(group D).In group C and group D,Co Cl₂ was added to the final concentration of 100?mol/l to make hypoxia environment.Then,GC-1 cells were cultured for 24h to establish hypoxia model.In group B and group D,emodin was added to the final concentration of 40?mol/l.CCK-8 assay and flow cytometry were applied to detect the level of apoptosis.Western blot and RT-q PCR were used to detect the expression of Bax and Caspase-3.Results:Compared with group A,CCK-8 assay showed the rate of cell survival was decreased in group C and group D.The rate of cell survival in group D was higher than in group C.The rate of cell apoptosis in group C increased compared with group A in the experiment of flow cytometry.However,The rate of cell apoptosis in group D was less than group C.Western blot and RT-q PCR showed that the level of expression of Bax and Caspase-3 in group C were significantly higher in contrast with group A.And the level of expression of Bax and Caspase-3 in group D decreased compared with group C.Conclusions:Emodin plays a protective role on spermatogonia in cell hypoxia model.The expression of Bax and Caspase-3 and the level of apoptosis decreased,the rate of cell survival increased by treatment with emodin.