| Sterility is both hotspot and nodus in andrology,however oligozoospermia and asthenospermia are the main reasons for sterility.As a commonly used drug for clinical treatment of oligospermia and asthenospermia,mechanism of Huang Jing Zan Yu capsule has attracted people's attention.Previous experiments showed that schisandrin A is an important component in Huang Jing Zan Yu capsule,which could promote the proliferation of interstitial cells.The aim of this study is to explore the mechanism of schisandrin A in oligospermia and asthenospermia in animal apoptosis.This thesis includes three sections:literature review,theoretical exploration and experimental research.Literature review systematically summarized the research progress of modern medicine on sterility.It summarize the pathogenesis and the clinical therapeutics.It probes into the current status and trend of the research on reproductive medicine,combs the historical evolution of the study of sterility in traditional Chinese medicine,and probes into the clinical clinical research and fundamental study.The application of traditional Chinese medicine in sterility is discussed,such as literature,experiment and epidemiology.The discussion contains three aspects:model,mechanism and drug.The model,pointcut and the reason for the choice of drug are introduced.The experimental section is divided into 4 sections.Objectives:1.Through the intervention of Schisandrin A in rat model of oligospermia and asthenospermia,the changes of semen parameters,histomorphology and sex hormones were observed,and the effect of schizandrin A on improving the model of oligospermia and asthenospermia was confirmed.2.Through molecular biology experiment technology,we explored the change of expression level of apoptosis related factors on SCF/c-kit pathway after the intervention of schisandrin A.3.Combining with the changes of schisandrin A on the histomorphological and molecular level,the apoptosis pathway is constructed.It provides a way to further explore the mechanism of schisandrin A to improve the mechanism of oligospermia and asthenospermia.Methods:Using glucoside tripterygium wilfordine(GTW)to produce 80 male SD rat models of oligospermia and asthenospermia,L-carnitine and Huang Jing Zan Yu capsule were selected as positive control drugs,and the models were made and administered for 35 days.After the end of the experiment,the semen index of rats was detected by sperm quality detection system.The levels of serum testosterone(T),luteinizing hormone(LH)and estradiol(E2)were detected by enzyme linked immunosorbent assay(ELISA).The pathological structure of testicular tissue was observed by optical microscope,and the microstructure was observed by electron microscope.The level of apoptosis-related factors SCF,c-kit,Bcl-2 and Caspase-3 expression were tested by immunohistochemistry(IHC)and Western Blot,and the mRNA expression level was tested by Real-time PCR.All final data were statistically analyzed using SPSS20.0.Result:1.After making the animal model,the semen parameters of the GTW group decreased significantly,and the quantity and quality of sperm in the group were changed,and the high-dose group of schisandrin A improved the sperm density and the spenn motility significantly(P<0.05).2-In terms of serum sex hormones,the GTW group caused dysfunction of the HPGA,decreased the T and E2 levels.Schizandrin A increased T level and adjusted LH and E2 levels.Schisandrin A in the low-dose group can effectively reduce the level of LH(P<0.05).3.Microscope showed that the seminiferous tubules in the GTW group were deformed,the spermatogenic cells fell off and disappeared,and the Sertoli cells atrophied.The high-dose group of schisandrin A was well constructed,the spermatogenic cells at all levels were arranged and the morphology was complete,and the morphology of Sertoli cells was basically normal.In the microstructural aspect,the swelling and vacuolation of the mitochondria and the abnormal increase of the autophagosome of the rats in the GTW group,and the morphologic changes of the organelles in the GTW group were slightly restored.4.The results of molecular biological experiments showed that GTW could down regulate the expression of anti apoptotic factor SCF,c-kit protein and mRNA,and up regulate the expression of Caspase-3 protein and mRNA(P<0.05).The high-dose group of schisandrin A increased the expression of SCF protein and mRNA(P<0.05).The high-dose group of schisandrin A could reduce the level of Caspase-3 protein and mRNA(P<0.05).The expression level of c-kit protein in each intervention group was larger than control group.The c-kit content in testicular tissue could be reduced by the middle-dose group of schisandrin A.Conclusion:Oligospermia and asthenospermia can reduce the number of spermatozoa and decrease the quality of sperm by changing the histomorphology of the testis,endocrine disorder,overexpression of apoptosis gene and inhibition of apoptosis genes,which eventually leads to sterility.Schisandrin A can improve the internal environment of the testis by restoring the morphology and function of the seminiferous tubules and up regulating the expression of anti apoptotic gene SCF to inhibit the abnormal apoptosis produced by SCF/c-kit apoptosis pathway,thus achieving the goal of spermatogenesis.Schisandrin A is one of the effective components of Huang Jing Zan Yu capsule.SCF/c-kit signal pathway can regulate the mechanism of cell apoptosis,proliferation and differentiation.We speculate that schisandrin A may interfere with the regulation of the proliferation of spermatogenic cells on SCF/c-kit pathway. |
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