Effect And Primary Mechanism Of Kallistatin Gene On Male Spermatogenesis

OBJECTIVE: By exploring the role of kallistatin gene in male spermatogenesis,the mechanism of kallistatin gene in spermatogenesis dysfunction was preliminarily explored,so as to find a new therapeutic target for spermatogenesis regulation and spermatogenesis dysfunction.METHODS: After obtaining the informed consent of the patients,the male semen samples were collected from the Department of reproductive medicine,the first people’s Hospital of Yunnan Province.The expression level of kalistatin in seminal plasma was detected by ELISA,and the relationship between the expression level of kalistatin in seminal plasma and sperm parameters was analyzed.The testis tissues of male infertility patients undergoing testicular puncture were collected and divided into normal control group,oligozoospermia group and non obstructive azoospermia group(10 cases in each group)The expression of kallstatin,tissue kallikrein-1(KLK1)and B2 receptor in testis and spermatozoa was detected by blotting,he staining and immunohistochemistry.The expression site and Johnsen score were evaluated by RT-PCR and western The level of apoptosis in testis was detected by blotting,the level of interstitial fibrosis in testis was detected by Masson staining and Sirius red staining.Finally,the relationship between the expression level of kallstatin protein in testis and spermatogenic function,the level of apoptosis and the level of tissue fibrosis was analyzed by correlation.RESULTS: The expression level of kalistatin in the seminal plasma of normal group,oligozoospermia group and non obstructive azoospermia group was in the order of normal group,oligozoospermia group and non obstructive azoospermia group(P < 0.001).The expression level of kalistatin in oligozoospermia group was positively correlated with sperm concentration.Kallstatin was highly expressed in the cytoplasm of spermatogenic cells,spermatozoa,Sertoli cells,stromal cells and peritubular myoid cells,which was similar to KLK1(P > 0.05).The expression level of kalistatin in the testis of normal group,oligozoospermia group and non obstructive azoospermia was in the order of normal group,oligozoospermia group and non obstructive azoospermia(P < 0.001),and there was a positive correlation between the expression level of kalistatin and Johnsen score.The level of apoptosis in testis of patients with non obstructive azoospermia and oligozoospermia was significantly higher than that of normal group(P < 0.05),and the expression level of Bcl-2 in non obstructive azoospermia and oligozoospermia group was significantly lower than that of normal group,followed by normal group > oligospermia > non obstructive azoospermia(P < 0.05),and Bax in non obstructive azoospermia group(P < 0.05)The expression levels of apoptosis protein,Bax / Bcl-2 and caspase-3 were significantly higher than those of the normal group and oligozoospermia group(P < 0.05).The expression level of apoptosis was negatively correlated with the expression level of kalistatin protein in testis.The level of fibrosis in testis tissue of patients with non obstructive azoospermia and oligospermia was significantly higher than that of the normal group(P<0.05).The order of the level was non obstructive azoospermia > oligospermia > normal group.The degree of fibrosis and the proportion of I/III collagen fiber were negatively correlated with the expression of kallistatin.Conclusion: Kalistatin has a significant correlation with male spermatogenesis.The decrease of kalistatin expression level may lead to spermatogenesis dysfunction.Kalistatin may participate in spermatogenesis regulation through anti apoptosis,anti fibrosis and other ways,and its regulatory mechanism needs to be further confirmed.