Effect Of Water Extract Of Cynomorium Songaricum On Sperm Motility,Serum Testosterone And The Proliferation Of Undifferentiated Spermatogonial Stem Cells In The Oligoasthenozoospermia Rat Model

objective:Increasing male infertility has been obserbed these years,however its etiology and pathology is still unclear.New efficiency drugs to treat the male infiertiliy are still needed.Male infertility is believed to associated with kidney dysfunction in the traditional Chinese medicine(TCM).The rise and fall of kidney Yin and Yang qi directly impact on male reproductive system.Cynomorium songaricum(CS)is used to improve sexual function and to treat kidney dysfunction in TCM.The purpose of this study is to investigate the effect of CS on sperm motility,serum testosterone and the proliferation and proliferation-associated gene expression of undifferentiated spermatogonial stem cells in the rat model of oligospermia and asthenospermia and to develop new traditional medicine drug.Mehtods:1.The 80 SPF SD male rats were randomly divided into four groups,the normal control group,the Model group,the CS treated group and the L-carnitine treated group.We use the intragastric administration of cytoxan to build the rat model of oligoasthenozoospermia,and then treat these oligoasthenozoospermia rats with CS or L-carnitine for four weeks by intragastric administration.2.The following parameters were observed:(1)The effect of CS on rat testis weight were observed.(2)We used HE staining to test the distribution of sperm cells in testicular tissue.(3)Computer-aided Sperm analysis(CASA)was used to detect the rat epididymal sperm tail number and vitality.(4)Chromatin diffusion method(SCD)was used to check the tail of epididymissperm DNA fragmentation index(DFI).(5)ELISA was used to detect concentration of FSH/T in rat serum.(6)ELISA was used to check the levels of lipid peroxidation(LPO)and reactive oxygen species(ROS)in epididymal sperm tail.(7)Fluorescence Activated Cell Sorter(FACS)was used to spermatagonial stem cells S phase proportion after CS processing.(8)Transferase-mediated deoxyuridine triphosphate-biotin nick end labeling(Tunel)staining method was used to detect apoptosis of ratspermatogonial stem cell lines.(9)Real-Time PCR method was used to detect the expression of spermatagonial stem Cells markers Thy-1 Cell Surface antigen(Thyl),GDNF-family receptor alpha-1(GFRa 1),promyelocytic leukemia zinc finger(PLZF)and C-kit mRNA in rat testicular tissue.Results and Conclusions:(1)After rats were treated with CS for four weeks,the results showed that the weight of testicular tissue of rat was slightly higher in CS group than the Model group.(2)HE staining results showed the layer number of seminiferous tubule in CS group obviously increased with more sperms in the lumen.(3)The CS group showed significant increases in epididymal sperm count compared to the Model group(P<0.05).(4)After 28 days of treatment with CS,the epididymal sperm motility were significantly higher compared to the Model group(P<0.05).(5)The epididymal sperm DNA fragmentation index(DFI)decreased in CS group compared to the Model group(P<0.05);The ROS level and LPO(lipid peroxidation)level decreased compared to the Model group(P<0.05).(6)The serum concentration of FSH and T increased in CS group compared with the Model Group(P<0.05).(7)S phase proportion of GFR alpha 1(+)spermatagonial stem cells increased after CS processing(P<0.05).(8)There was no change in apoptosis of SSC between the two groups.(9)The Thy1,GFR alpha,PLZF gene mRNA level of rat testicular tissue increased(P<0.05)in CS group.But there was no significant difference in C-kit gene expression.(10)CS also increased the expression of GDNF compared to the Model group.These results suggest that CS may improve male fertility by enhancing spermatogenesis and improving motility and quanitity of epididymal sperm.