Study The Effects Of Cyclosporin A On Spermatogenesis And Expression Of Fasl And Fas In The Controlateral Testis After The Unilateral Testis Injured In KM Mouse

Objective: To investigate the effects of CsA (Cyclosporin A) on spermgenesis and expression of FasL and Fas in the contra lateral testis after the unilateral testis was injured by glacial acetic acid in KM mouse.Methods: 1. A total of 60 mice were randomly divided into groups A (normal control), B (the right testis injured by glacial acetic acid), C (excision of ipsilateral testis after the right testis injured by glacial acetic acid at 6 hours) and D (Administered with 10mg/kg of CsA for 4 consecutive weeks after the right testis injured by glacial acetic acid) equally. 2. All of KM mice were exsanguinated from the post-ocular vein after 4 weeks in order to collect serum. Antibodies in serum were assayed by SP method. 3. Meanwhile all the left epididymides were removed from mice and squeezed with plunger in 2 ml Hank's balanced salt solution (HBSS) at the temperature of 37℃. After incubated 30 minutes sperm staining with trypan blue were counted through microscope and sperm density and spermmotility were evaluated. 4. The left testes were fixed in NBF about 48 hours, then in Bouin's solution about 18 hours and embedded in paraffin using standard procedures. Light and electron microscope of the left testes analyses were conducted. 5. Expression of FasL and Fas were performed by immunohistochemistry (SP method). The positive cells with SP staining in seminiferous tubules were calculated. For statistical evaluation one-way ANOVA were used by SPSS10.0 for windows. A p value less than 0.05 was taken as significant. Results: 1.There were the motile sperms of all groups in HBSS at the temperature of 37℃ although the viability counting by the trypan blue method decreased and many sperms were dead in group B. Sperm density and sperm motility in group B were significantly decreased compared with another groups (P<0.05). 2. Antibodies on anti-germ and anti-basal membranes in serum were found in different degree by SP method in group B, C and D. And there was a lack of CD3 positive cells. 3.The vacuolar degeneration was observed in Sertoli cells and spermatogenic cells through electron microscope and the total of testicular cells in average seminiferous tubule decreased through light microscope. There were a few germs and some shedding testicular cells in the left ductus epididymides after the right testis were injured by glacial acetic acid. Those that administered with 10mg/kg of CsA took a favorable turn. 4. There were some seminiferous tubules of FasL and Fas protein positive and negative immunoreactions. Some spermatogenic cells were FasL and Fas protein positive immunoreactions on control that all of the Sertoli cells were always negative immunoreactions. Expression of FasL and Fas in group B increasedsignificantly compared with control group (P<0.05). Expression of FasL and Fas in group D decreased significantly compared with group B (P<0.05).Conclusions: 1. Sperm density and sperm motility in group B were significantly decreased and those that administered with 10mg/kg of CsA would improve. 2. Antibodies on anti-germ and anti-basal membranes in serum changed the environment of spermatogenesis and lymphocytes might have no effect. 3.Expressions of FasL and Fas in testis were due to some spermatogenic cells rather than to the Sertoli cells. The seminiferous tubules at stagesⅪ-Ⅻ had expression of FasL and Fas in all groups. The tubules beyond stagesⅪ-Ⅻ expressed FasL and Fas in group B. Fas system controls not only apoptosis of physiological germ cell but also apoptosis of germ cell induced by this injury. 4. At certain concentration Cyclosporin A decreased expression of Fas and FasL and maintained spermatogenesis of the contralateral testis after the unilateral testis was injured.