Follicle-stimulating hormone autoantibody is involved in idiopathic spermatogenic dysfunction

Aim： To detect the anti-follicle-stimulating hormone （FSH） antibody in idiopathic infertile patients and fertile subjects in order to determine the role of this antibody in patients with spermatogenic dysfunction. Methods： The anti-FSH antibody in serum was detected by an enzyme-linked immunosorbent assay （ELISA）. The functional and structural integrity of the sperm membrane was evaluated with hypo-osmotic swelling （HOS） test and the ultrastructure of the spermatozoa was investigated by transmission electron microscopy （TEM）. Results： The extent of positive FSH antibody in the patients with oligozoospermia and/or asthenozoospermia was significantly higher than that in the fertile subjects and infertile patients with normal sperm concentration and motility, but it was significantly lower than that in the patients with azoospermia. The extent of anti-FSH antibody in the patients with azoospermia was significantly greater than that in patients with oligospermia and/or asthenospermia, infertile people with normal sperm density and motility and fertile people. The hypo-osmotic swelling test showed that the percentage of HOS-positive spermatozoa （swollen） was 45.1% ±3.5% in the FSH antibody-positive group and 59.1% ± 6.2% in the FSH antibody-negative control group. The percentage of functional membrane damage to spermatozoa was significantly higher in the anti- FSH antibody-positive group than in the control group. TEM showed that the outer acrosomal membrane was located far from the nucleus, and detachment of the acrosome was found in the FSH autoantibody-positive group. Conclusion： These data suggest that the presence of anti-FSH antibody is strongly correlated with the sperm quantity and quality in idiopathic male infertility. Anti-FSH antibody may be an important factor causing spermatogenic dysfunction and infertility.

Network Pharmacology-based Strategy for Predicting Active Ingredients and Potential Targets of Cuscutae semen and Lycii fructus in Spermatogenic Dysfunction

Background:Traditional Chinese Medicine(TCM)Cuscutae semen(SC,Tusizi from Cuscuta chinensis Lam.)and Lycii fructus(FL,Gouqi from Lycium barbarum L.),are also used as a herb pair(SC-FL)to treat various ailments,including spermatogenic dysfunction(SD),a disease responsible for low fertility in males.Objective:Herein,we will further determine the bioactive components,component targets and partial molecular mechanisms of the herb pair for the treatment of SD.Methods:We employed the traditional Chinese medicine systems pharmacology database in combination with Ultra high performance liquid chromatography(UHPLC)analysis to analyze the active ingredients of the SC-FL herbal pair and explore its possible targets and underlying mechanism in treatment of spermatogenic dysfunction(SD).Moreover,we used a Sprague-Dawley rat model,generated by using glucosides of Tripterygium wilfordii(GTW),to evaluate the effect of SC-FL and the underlying mechanism on SD and reliability of certain key targets and pathways obtained from the pharmacology analysis.Results:We identified 56 active ingredients in SC-LF affecting 41 overlapping gene signals that influenced SD treatment outcomes.262 Gene Ontology(GO)terms and 170 pathways were yielded under analyses of Gene Ontology,Kyoto Encyclopedia of Gene and Genome pathway from which we predicted that cell proliferation and apoptosis were the primary biological processes(BP)involved in the treatment of SD by SC-LF.The results showed that SC-FL treatment significantly improved the testicular organ coefficient along with sperm count and motility,while reduced testicular damage and testicular tissue cell apoptosis in SD model.Mechanistically,SC-FL significantly upregulated the expression levels of anti-apoptotic proteins AR and BCL2 and downregulated those of pro-apoptotic proteins BAD,BAX,cleaved caspase 3,and CASP3.Conclusion:Collectively,these results indicated that SC-FL elicited a protective effect by potentially regulating apoptosis,thus suggesting that it will represent an effective reagent for male infertility.

Apoptotic and nonapoptotic function of caspase 7 in spermatogenesis

Recent studies have reported that caspase 7 has an apoptotic and nonapoptotic function. However, the relationship between caspase 7 and spermatogenesis remains unknown. This study aimed to investigate the possible function of caspase 7 during normal and abnormal spermatogenesis. The cleaved form of caspase 7 was detected in testis tissues at different postpartum times （5-14 weeks） by qRT-PCR, Western blot and immunohistochemistry （IHC）. Then, the mice models of spermatogenic dysfunction were obtained by busulfan （30 mg kg-1） to further evaluate the potential function and mechanism of caspase 7. qRT-PCR and Western blot results showed that caspase 7 expression was gradually elevated from 5 to 14 weeks, which was not connected with apoptosis. IHC results revealed that caspase 7 was mainly located in spermatogenic cells and Leydig cells. In addition, spermatogenic dysfunction induced by busulfan gradually enhanced the apoptosis and elevated the expression of caspase 3, caspase 6, and caspase 9, but decreased the expression of caspase 7 in spermatogenic cells. However, when spermatogenic cells were mostly disappeared at the fourth week after busulfan treatment, caspase 7 expression in Leydig cells was significantly increased and positively correlated with the expression of caspase 3, caspase 6, and caspase 9. Therefore, these results indicate that caspase 7 has a nonapoptic function that participates in normal spermatogenesis, but also displays apoptotic function in spermatogenic dysfunction.