PRSS50 is a testis protease responsible for proper sperm tail formation and function.

Multiple morphological abnormalities of the sperm flagella (MMAF) are a major cause of asthenoteratozoospermia. We have identified protease serine 50 (PRSS50) as having a crucial role in sperm development, because Prss50-null mice presented with impaired fertility and sperm tail abnormalities. PRSS50 could also be involved in centrosome function because these mice showed a threefold increase in acephalic sperm (head-tail junction defect), sperm with multiple heads (spermatid division defect) and sperm with multiple tails, including novel two conjoined sperm (complete or partial parts of several flagellum on the same plasma membrane). Our data support that, in the testis, as in tumorigenesis, PRSS50 activates NFκB target genes, such as the centromere protein leucine-rich repeats and WD repeat domain-containing protein 1 (LRWD1), which is required for heterochromatin maintenance. Prss50-null testes have increased IκκB, and reduced LRWD1 and histone expression. Low levels of de-repressed histone markers, such as H3K9me3, in the Prss50-null mouse testis may cause increases in post-meiosis proteins, such as AKAP4, affecting sperm formation. We provide important insights into the complex mechanisms of sperm development, the importance of testis proteases in fertility and a novel mechanism for MMAF.

Upregulated microRNA-423-5p promotes oxidative stress through targeting glutathione S-transferase mu 1 in asthenozoospermia.

The dysregulation of microRNAs (miRNAs) plays an important role in asthenozoospermia. This study evaluated the sperm microRNA-423-5p (miR-423-5p) expression in asthenozoospermia and normozoospermia, exploring the role of miR-423-5p in asthenozoospermia. Eighty participants were divided into asthenozoospermic (AZS, n = 40) and normozoospermic (Norm, n = 40) groups. Fresh semen samples were collected and the sperm cells were separated. Quantitative Real-Time polymerase chain reaction was used to measure the sperm miR-423-5p level. Receiver operating characteristic curve (ROC) was employed to test the diagnostic performance of miR-423-5p in asthenospermia. Dual-reporter luciferase assay was adopted to confirm the target gene of miR-423-5p. The target gene level in asthenozoospermia and normozoospermia was measured, and the biological function of target gene in asthenozoospermia was evaluated. Results showed that the miR-423-5p expression level in the AZS group was higher than that in Norm group, which was positively correlated with the severity of asthenozoospermia. ROC analysis of miR-423-5p showed an area under curve (AUC) of 0.69 (95% confidence interval = 0.57-0.80, p <0 .01), with 80% sensitivity and 60% specificity. Glutathione S-transferase mu 1 (GSTM1) is a target gene of miR-423-5p, which significantly decreased in the AZS group. Compared with Norm group, glutathione S-transferase (GST) activity and total antioxidant capacity (TAC) level decreased, while malondialdehyde (MDA) level increased in the AZS group. Furthermore, GST activity and TAC level were negatively correlated with miR-423-5p expression, while MDA level was positively correlated with miR-423-5p expression. In conclusion, the sperm miR-423-5p level significantly was upregulated in asthenozoospermia. High-level miR-423-5p inhibited sperm motility through targeting GSTM1 to promote oxidative stress.

Acetylation/deacetylation and microtubule associated proteins influence flagellar axonemal stability and sperm motility.

PTMs and microtubule-associated proteins (MAPs) are known to regulate microtubule dynamicity in somatic cells. Reported literature on modulation of α-tubulin acetyl transferase (αTAT1) and histone deacetylase 6 (HDAC6) in animal models and cell lines illustrate disparity in correlating tubulin acetylation status with stability of MT. Our earlier studies showed reduced acetyl tubulin in sperm of asthenozoospermic individuals. Our studies on rat sperm showed that on inhibition of HDAC6 activity, although tubulin acetylation increased, sperm motility was reduced. Studies were therefore undertaken to investigate the influence of tubulin acetylation/deacetylation on MT dynamicity in sperm flagella using rat and human sperm. Our data on rat sperm revealed that HDAC6 specific inhibitor Tubastatin A (T) inhibited sperm motility and neutralized the depolymerizing and motility debilitating effect of Nocodazole. The effect on polymerization was further confirmed in vitro using pure MT and recHDAC6. Also polymerized axoneme was less in sperm of asthenozoosperm compared to normozoosperm. Deacetylase activity was reduced in sperm lysates and axonemes exposed to T and N+T but not in axonemes of sperm treated similarly suggesting that HDAC6 is associated with sperm axonemes or MT. Deacetylase activity was less in asthenozoosperm. Intriguingly, the expression of MDP3 physiologically known to bind to HDAC6 and inhibit its deacetylase activity remained unchanged. However, expression of acetyl α-tubulin, HDAC6 and microtubule stabilizing protein SAXO1 was less in asthenozoosperm. These observations suggest that MAPs and threshold levels of MT acetylation/deacetylation are important for MT dynamicity in sperm and may play a role in regulating sperm motility.

DAAM1 and PREP are involved in human spermatogenesis.

During differentiation of the male gamete, there is a massive remodelling in the shape and architecture of all the cells in the seminiferous epithelium. The cytoskeleton, as well as many associated proteins, plays a pivotal role in this process. To better characterise the factors involved, we analysed two proteins: the formin, dishevelled-associated activator of morphogenesis 1 (DAAM1), which participates in the regulation of actin polymerisation, and the protease, prolyl endopeptidase (PREP), engaged in microtubule-associated processes. In our previous studies we demonstrated their involvement in cytoskeletal dynamics necessary for correct postnatal development of the rat testis. Here, we used samples of testicular tissue obtained from infertile men by testicular sperm extraction and the spermatozoa of asthenoteratozoospermic patients. By western blot and immunofluorescent analysis, we found that DAAM1 and PREP expression and localisation were impaired in both the testis and spermatozoa, and in particular in the midpiece as well as in the principal and end-pieces of the flagella, as compared with spermatozoa of normospermic men. Our results provide new knowledge of the dynamics of spermatogenesis, raising the possibility of using DAAM1 and PREP as new markers of normal fertility.

Proteomics analysis of asthenozoospermia and identification of glucose-6-phosphate isomerase as an important enzyme for sperm motility.

Asthenozoospermia, in which sperm motility is affected, is one of the primary causes of male infertility. However, the exact mechanism responsible for the defective motility remains unknown. It is important to identify the precise proteins or pathways involved in sperm motility. The present study analyzed five asthenozoospermic sperm samples and five healthy controls using TMT-based quantitative method and identified 152 differentially expressed proteins, with 84 upregulated and 68 downregulated in asthenozoospermia. Four proteins (GPI, MDH1, PGAM1 and PGAM2) were found in several over-represented energy metabolism pathways using bioinformatics analysis. Glucose-6-phosphate isomerase (GPI), a rate-limiting enzyme converting glucose-6-phosphate to fructose-6-phosphate, was found to be significantly decreased in asthenozoospermia by Western blotting and ELISA on an extended sample size. Furthermore, substitution of glucose with fructose-6-phosphate significantly promoted asthenozoospermic sperm motility in vitro. Taken together, our results suggest that the poor motility of sperm in asthenozoospermia may partly result from defects in GPI-associated energy metabolism. SIGNIFICANCE: To identify the key proteins or pathways involved in sperm motility, the accurate TMT-based quantitative method was applied to characterize protein profiles of asthenozoospermic sperm. GPI, an enzyme involved in energy metabolism, was found to be differentially abundant, and validated by extended sample analysis. The supplement of the product of GPI, fructose-6-phosphate, could significantly improve sperm motility. Our study could provide new insights into the molecular basis of sperm motility and the improvement of motility in asthenozoospermia.

Isoform-specific GSK3A activity is negatively correlated with human sperm motility.

In mouse and bovine sperm, GSK3 activity is inversely proportional to motility. Targeted disruption of the GSK3A gene in testis results in normal spermatogenesis, but mature sperm present a reduced motility, rendering male mice infertile. On the other hand, GSK3B testis-specific KO is fertile. Yet in human sperm, an isoform-specific correlation between GSK3A and sperm motility was never established. In order to analyze GSK3 function in human sperm motility, normospermic and asthenozoospermic samples from adult males were used to correlate GSK3 expression and activity levels with human sperm motility profiles. Moreover, testicular and sperm GSK3 interactomes were identified using a yeast two-hybrid screen and coimmunoprecipitation, respectively. An extensive in-silico analysis of the GSK3 interactome was performed. The results proved that inhibited GSK3A (serine phosphorylated) presents a significant strong positive correlation (r = 0.822, P = 0.023) with the percentage of progressive human sperm, whereas inhibited GSK3B is not significantly correlated with sperm motility (r = 0.577, P = 0.175). The importance of GSK3 in human sperm motility was further reinforced by in-silico analysis of the GSK3 interactome, which revealed a high level of involvement of GSK3 interactors in sperm motility-related functions. The limitation of techniques used for GSK3 interactome identification can be a drawback, since none completely mimics the physiological environment. Our findings prove that human sperm motility relies on isoform-specific functions of GSK3A within this cell. Given the reported relevance of GSK3 protein-protein interactions in sperm motility, we hypothesized that they stand as potential targets for male contraceptive strategies based on sperm motility modulation.

[Expressions of the adenylate kinase family in asthenospermia].

OBJECTIVE: To investigate the expressions of the adenylate kinase (AK) family in the sperm of asthenospermia patients. METHODS: We collected semen samples from 30 asthenospermia patients and another 30 normal healthy males, detected the mRNA and protein expressions of AKs by RT-PCR and Western blot, localized the AKs by immunofluorescence assay and immunohistochemistry, and analyzed their association with sperm motility. RESULTS: RT-PCR and Western blot showed that AKs 1－9 were expressed in the sperm of all the subjects, and the mRNA and protein expressions of AK1, AK6 and AK7 were significantly lower in the asthenospermia patients than in the normal males. In the testis, AK1 and AK7 were expressed in the cytoplasm while AK6 in the nucleus, and in the sperm, the former two were localized in the flagella while the latter one in the sperm head. Antibody blocking experiments showed that none of the AK1, AK6 and AK7 antibodies had any significant effect on the total or progressive motility of the sperm while cultured with each alone, but co-culturing with the three antibodies markedly reduced the total sperm motility (ï¼»80.5 ± 2.4ï¼½%) and progressive sperm motility (ï¼»60.6 ± 3.6ï¼½%) in comparison with those in the control group (ï¼»87.6 ± 3.3ï¼½% and ï¼»70.2 ± 2.3ï¼½%) (P < 0.05). CONCLUSIONS: The mRNA and protein expressions of AK1, AK6 and AK7 are significantly down-regulated in the sperm of asthenospermia patients, which may be closely related with reduced sperm motility.

[Effect of Qiangjing Tablets on the MAPK signaling pathway in SD rats with asthenospermia].

OBJECTIVE: To investigate the effects of Qiangjing Tablets (QJT) on sperm quality and the MAPK signaling pathway in the SD rat model of asthenospermia (AS). METHODS: A total of 100 male SD rats were randomly divided into five groups of equal number, blank control, AS model control, high-dose QJT, medium-dose QJT, and low-dose QJT. All the rats were intragastrically administered ORN at 200 mg/kg/d for establishment of the AS model except those in the blank control group, which were given 1% CMC sodium solution at 1 ml/100 g by gavage. Meanwhile the animals of the high-, medium-, and low-dose QJT groups were gavaged with QJT at 6700, 3300 and 1700 mg/kg/d, respectively, qd 6 days a week for 20 days. Then the testis issue and the apoptosis of the testicular cells were observed under the electron microscope, the expression of vimentin in the testis was determined with the immunohistochemical SP method, that of ERK1/2 detected by Western blot, and the concentration of TGF-ß1 in the semen measured by ELISA. RESULTS: The AS model controls showed round nuclei of spermatocytes, homogeneously distributed chromatins, broken or lost mitochondria, and expanded rough endoplasmic reticulum in the testis tissue. In comparison, the rats of the high-, medium-, and low-dose QJT groups exhibited round nuclei of spermatocytes, homogeneously distributed chromatins, and well-structured mitochondria, rough endoplasmic reticulum and ribosome, which were all similar those of the blank controls. Compared with the blank controls, the AS model rats manifested significantly increased expressions of ERK1/2 (1.00 ± 0.00 vs 1.26 ± 0.10, P<0.01) and vimentin (0.16 ± 0.01 vs 0.17 ± 0.01, P<0.01) and apoptosis rate of cells in the testis tissue (ï¼»9.20 ± 3.07ï¼½ vs ï¼»42.20 ± 9.17ï¼½ %, P<0.01), but decreased level of TGF-ß1 in the semen (ï¼»627.67 ± 26.07ï¼½ vs ï¼»566.73 ± 68.44ï¼½ ng/ml, P<0.05). In comparison with the model controls, the rats of the high- and medium- -dose QJT groups presented remarkably down-regulated expressions of ERK1/2 (1.26 ± 0.10 vs 1.14 ± 0.08, P<0.01; 1.26 ± 0.10 vs 1.18 ± 0.05, P<0.05) and vimentin (0.17 ± 0.01 vs 0.16 ± 0.01, P<0.01; 0.17 ± 0.01 vs 0.17 ± 0.09, P<0.05) and decreased rate of cell apoptosis (ï¼»42.20 ± 9.17ï¼½ vs ï¼»21.60 ± 5.94ï¼½ %, P<0.01; ï¼»42.20 ± 9.17ï¼½ vs ï¼»33.95 ± 6.39ï¼½ %, P<0.05). The concentration of TGF-ß1 in the semen was markedly lower in the high-dose QJT than in the AS model control group (ï¼»621.78 ± 30.80ï¼½ vs ï¼»566.73 ± 68.44ï¼½ ng/ml, P < 0.05). CONCLUSIONS: Qiangjing Tablets could improve semen quality in asthenospermia rats by acting against oxidative stress.

Plasma membrane calcium ATPase 4 (PMCA4) co-ordinates calcium and nitric oxide signaling in regulating murine sperm functional activity.

Reduced sperm motility (asthenospermia) and resulting infertility arise from deletion of the Plasma Membrane Ca2+ -ATPase 4 (Pmca4) gene which encodes the highly conserved Ca2+ efflux pump, PMCA4. This is the major Ca2+ clearance protein in murine sperm. Since the mechanism underlying asthenospermia in PMCA4's absence or reduced activity is unknown, we investigated if sperm PMCA4 negatively regulates nitric oxide synthases (NOSs) and when absent NO, peroxynitrite, and oxidative stress levels are increased. Using co-immunoprecipitation (Co-IP) and Fluorescence Resonance Energy Transfer (FRET), we show an association of PMCA4 with the NOSs in elevated cytosolic [Ca2+ ] in capacitated and Ca2+ ionophore-treated sperm and with neuronal (nNOS) at basal [Ca2+ ] (ucapacitated sperm). FRET efficiencies for PMCA4-eNOS were 35% and 23% in capacitated and uncapacitated sperm, significantly (p < 0.01) different, with the molecules being <10 nm apart. For PMCA4-nNOS, this interaction was seen only for capacitated sperm where FRET efficiency was 24%, significantly (p < 0.05) higher than in uncapacitated sperm (6%). PMCA4 and the NOSs were identified as interacting partners in a quaternary complex that includes Caveolin1, which co-immunoprecipitated with eNOS in a Ca2+ -dependent manner. In Pmca4-/- sperm NOS activity was elevated twofold in capacitated/uncapacitated sperm (vs. wild-type), accompanied by a twofold increase in peroxynitrite levels and significantly (p < 0.001) increased numbers of apoptotic germ cells. The data support a quaternary complex model in which PMCA4 co-ordinates Ca2+ and NO signaling to maintain motility, with increased NO levels resulting in asthenospermia in Pmca4-/- males. They suggest the involvement of PMCA4 mutations in human asthenospermia, with diagnostic relevance.

Sperm Na+, K+-ATPase α4 and plasma membrane Ca2+-ATPase (PMCA) 4 regulation in asthenozoospermia.

Asthenozoospermia, which is characterized by reduced motility, is one of the etiologies of male infertility. Its biochemical and functional consequences include altered ATPase activity. This study investigated the activities of Na+, K+-ATPase and Ca2+-ATPase and the expression of Na+, K+-ATPase α4 and PMCA4 isoforms in human sperm of asthenozoospermic infertile men. Nineteen samples from asthenozoospermic infertile couples were examined in this study. Computerized-assisted semen analysis (CASA) was performed, and the enzyme activity was measured based on the ability of ATPase to release organic phosphate from ATP as a substrate. The Na+, K+-ATPase α4 and PMCA4 isoform expression levels were measured by western immunoblotting, whereas the protein distribution was examined by immunocytochemistry. This showed that the Na+, K+-ATPase activity and the Na+, K+-ATPase α4 isoform expression were lower in the asthenozoospermia group than in the normozoospermia group (8.688±1.161 versus 13.851±1.884 µmol Pi/mg protein/h, respectively; p>0.05). In contrast, the Ca2+-ATPase activity was significantly higher in the asthenozoospermia group than in the normozoospermia group (11.154±1.186 versus 2.725±0.545 µmol Pi/mg protein/h, respectively; p<0.05). In comparison, PMCA4 expression in the asthenozoospermia group was lower than in the normozoospermia group (p>0.05). The altered ATPase activity and isoform expression in asthenozoospermia may impair sperm structure and function.

Sperm glyceraldehyde 3-phosphate dehydrogenase gene expression in asthenozoospermic spermatozoa.

It has been suggested that the energy required for sperm motility is produced by oxidative phosphorylation while glycolysis seems to be an important source for ATP transmission along the flagellum. Some studies have investigated the chemical and kinetic properties of the enzyme glyceraldehyde 3-phosphate dehydrogenase to identify any changes in the regulation of glycolysis and sperm motility. In contrast, there are few studies analyzing the genetic basis of hypokinesis. For this reason, we investigated the glyceraldehyde 3-phosphate dehydrogenase gene in human sperm to evaluate whether asthenozoospermia was correlated with any changes in its expression. Semen examination and glyceraldehyde 3-phosphate dehydrogenase gene expression studies were carried out on 116 semen samples divided into two groups - Group A consisted of 58 normokinetic samples and Group B of 58 hypokinetic samples. Total RNA was extracted from spermatozoa, and real-time PCR quantification of mRNA was carried out using specific primers and probes. The expression profiles for the Groups A and B were very similar. The mean delta Ct was as follows - Group A, 5.79 ± 1.04; Group B, 5.47 ± 1.27. Our study shows that in human sperm, there is no difference in glyceraldehyde 3-phosphate dehydrogenase gene expression between samples with impaired motility and samples with normal kinetics. We believe that this study could help in the understanding of the molecular mechanisms of sperm kinetics, suggesting that hypomotility may be due to a possible posttranscriptional impairment of the control mechanism, such as mRNA splicing, or to posttranslational changes.

Ubiquitin Carboxy-Terminal HydrolaseL3 Correlates with Human Sperm Count, Motility and Fertilization.

Ubiquitin C-terminal hydrolase L3 (UCHL3) belongs to the group of deubiquitinating enzymes and plays a part in apoptosis of germ cells and the differentiation of spermatocytes into spermatids. However, the exact role of UCHL3 in human spermatogenesis and sperm function remains unknown. Here we examined the level and activity of UCHL3 in spermatozoa from men with asthenozoospermia (A), oligoasthenozoospermia (OA) or normozoospermia (N). Immunofluorescence indicated that UCHL3 was mainly localized in the acrosome and throughout the flagella, and western blotting revealed a lower level in A or OA compared with N (p < 0.05). The catalytic activity of UCHL3 was decreased in spermatozoa from A or OA (p < 0.05, p < 0.001, respectively). The level and activity of UCHL3 were positively correlated with sperm count, concentration and motility. The UCHL3 level was positively correlated with the normal fertilization rate (FR) and percentage of embryos suitable for transfer/cryopreservation of in vitro fertilization (IVF). The UCHL3 activity was also positively correlated with FR, the percentage of embryos suitable for transfer/cryopreservation and high-quality embryos rate of IVF. Aforementioned correlations were not manifested in intra-cytoplasmic sperm injection (ICSI). These findings suggest that UCHL3 may play a role in male infertility.

THE PECULIARITIES OF ARGINASE PATHWAY OF L-ARGININE IN SPERMATOZOA IN MEN WITH DIFFERENT FORMS OF PATHOSPERMIA.

The changes in arginase activity of spermatozoa and hormonal profile of peripheral blood of infertile men with various forms pathospermia have been studied. It has been found that arginase activity in the sperm cells of men with oligozoo-, antenozoo-, oligoastenozoo- and leucocytospermia is decreased in 2.1, 2.3, 2.4 and 3.3 times respectively. This indicates about inhibition of arginase pathway of L-arginine metabolism, which is not significantly dependent on the type of disruption of spermatogenesis. The most significant changes have been observed in infertile men with leucocytospermia since white blood cells stimulate the formation of reactive oxygen species, induction and development of oxidative and nitrative stress in spermatozoa. Inhibition of arginase pathway of L-arginine metabolism has adaptive role, which is to limit bioavailabil- ity of L-arginine and to prevent excessive formation of NO in cytotoxic concentrations to sperm cells. It has been noted changes in serum concentrations of gonadotropin and sex hormones in men with various forms of pathospermia. The most expressed significant changes were in levels of follicle stimulating hormone and testosterone. The concentration of follicle stimulating hormone in patients with oligozoospermia caused by hypogonadism is twice higher and in patients with leucocytospermia in 1.8 times higher than in fertile men. In patients with astenozoospermia this value is in 2.2 times lower than in normozoospermic samples but within the physiological norm. The testosterone level in men with oligozoospermia is in 1.6 times lower than in fertile men but within the physiological norm. It has been found that arginase inhibition of spermatozoa po6itively correlated with a decrease in their concentration in the ejaculate of infertile men with oligozoospermia (r =0.68).

Seminal cyclooxygenase relationship with oxidative stress in infertile oligoasthenoteratozoospermic men with varicocele.

This study aimed to assess the relation of seminal cyclooxygenase COX-1, COX-2 with oxidative stress in infertile oligoasthenoteratozoospermic (OAT) men with varicocele (Vx). In all, 128 men were allocated into fertile men, fertile men with Vx, infertile OAT men without Vx and infertile OAT men with Vx. They were subjected to history taking, clinical examination and semen analysis. Also, seminal COX-1, COX-2, malondialdehyde (MDA) and glutathione peroxidase (GPx) were estimated. Mean levels of seminal COX-1, COX-2 were over-expressed, the mean level of seminal MDA was significantly increased, and the mean level of seminal GPx was significantly decreased in infertile OAT men with Vx compared with other groups. Seminal COX-1 and COX-2 were over-expressed in cases with Vx grade III compared with Vx grades I, II cases and in cases with bilateral Vx compared with unilateral Vx. There was significant negative correlation between seminal COX-1 and COX-2 with sperm concentration, sperm motility, sperm normal morphology, seminal GPx and significant positive correlation with seminal MDA. It is concluded that seminal COX-1 and COX-2 are over-expressed in infertile OAT men with Vx compared with fertile men with/without and infertile OAT men without Vx being associated with oxidative stress, Vx grade and Vx laterality.

Asthenozoospermia and membrane remodeling enzymes: a new role for phospholipase A2.

Phosholipase A2 (PLA2 ) activity in the seminal plasma and in sperm heads is closely related to sperm motility and male fertility. Therefore, the purpose of this study was to investigate the possible involvement of different isoforms of phospholipase in asthenozoospermia. To accomplish this, cPLA2 , phospho-cPLA2 , iPLA2 , and sPLA2 were evaluated by immunofluorescence and immunoblot analyses in spermatozoa obtained from 22 normozoospermic men and 28 asthenozoospermic patients. We found significant differences in cPLA2 and its phosphorylated/activated form, iPLA2 , and sPLA2 content and distribution in normal and asthenozoospermic patients. cPLA2 was localized in heads, midpieces, and tails of all spermatozoa as constitutive enzyme, less expressed in the tail of spermatozoa with low progressive motility. While active phospho-cPLA2 distribution was homogeneous throughout the cell body of control-donor spermatozoa, lower levels were detected in the tails of asthenozoospermic patients, as opposed to its strong presence in heads. Low immunofluorescence signal for iPLA2 was found in astenozoospermic patients, whereas sPLA2 was significantly lower in the heads of asthenozoospermic patients. Spermatozoa with low progressive motility showed differences both in terms of total specific activity and of intracellular distribution. cPLA2 , iPLA2 , and sPLA2 specific activities correlated positively and in a significantly manner with sperm progressive motility both in normozoospermic men and asthenozoospermic patients. In conclusion, PLA2 s are expressed in different areas of human spermatozoa. Spermatozoa with low motility showed differences in total specific activity and enzyme distributions. We speculated that PLA2 expression and/or different distribution could be potential biomarkers of asthenozoospermia, one of the major causes of male factor infertility.

[Shugan Yiyang Capsules for the treatment of asthenospermia: A clinical study].

OBJECTIVE: To observe the clinical efficacy of Shugan Yiyang Capsules in the treatment of asthenospermia and its action mechanisms. METHODS: We randomly assigned 135 asthenospermia patients to groups A (n = 47), B (n = 45), and C (n = 43) to be treated with Shugan Yiyang Capsules, oral levocarnitine, or combination of the two. We observed sperm quality and the level of α-glucosidase in the seminal plasma before and after medication. RESULTS: The total effectiveness rate was 70.21% in group A (markedly effective in 16 cases and effective in 17), 68.89% in group B (markedly effective in 15 cases and effective in 16), and 83.72% in group C (markedly effective in 16 cases and effective in 20), significantly higher in C than in A and B (P < 0.05). Both sperm quality and the level of α-glucosidase in the seminal plasma were improved in the three groups of patients, most obviously in group C. CONCLUSION: Shugan Yiyang Capsules can be used for the treatment of asthenospermia, and its effect can be enhanced in combination with oral levocarnitine.

Light-emitting diode exposure enhances sperm motility in men with and without asthenospermia: preliminary results.

OBJECTIVE: To evaluate the effect of light-emitting diode (LED) on sperm motility in men with and without asthenospermia. MATERIAL AND METHODS: Semen samples from 27 men were assessed and washed. An aliquot was taken from each sample as a control. The remaining amount was exposed to red LED for 2, 5 and 10 minutes. Sperm motility from the test and control tubes were re-checked at the end of each time interval. In 11 of these 27 samples, the same protocol was repeated without sperm washing. Evaluation of sperm creatine kinase (CK) activity, hypoosmotic swelling (HOS) test and aniline blue staining (ANBS) were undertaken after phototherapy in additional 15 samples. RESULTS: Progressive sperm motility increased significantly after LED treatment at the different time intervals whether in washed (p = 0.000) or non-washed (p = 0.003) samples. The amount of the increase in motility in washed aliquots was significantly more (p = 0.000) than in naive semen. Sperm CK activity increased, but was not significant whilst there were no changes regarding HOS and ANBS. CONCLUSION: Red LED is a promising safe tool to boost sperm motility in vitro. This may have a great implication on maximizing the possibilities and outcomes of intrauterine insemination trials.

Differential expression of mRNA aromatase in ejaculated spermatozoa from infertile men in relation to either asthenozoospermia or teratozoospermia.

Oestrogen biosynthesis in ejaculated spermatozoa is an autonomous process, which may influence sperm functions. The purpose of this study was to evaluate the relationship between the expression of aromatase, sperm quality and seminal neutral α-glucosidase marker in semen of Tunisian infertile men: asthenozoospermia (A; n = 16), teratozoospermia (T; n = 12) and asthenoteratozoospermia (AT; n = 11) in comparison with 18 normozoospermic ones. Aromatase mRNA levels estimated by real-time PCR were reduced in groups T (52%) and AT (67%) compared to controls and inversely correlated with the percentage of normal forms. A higher coefficient of correlation was noted in presence of microcephaly or acrosome malformations (r = -0.64). The asthenozoospermic group was divided into two subgroups according to the relative amount of aromatase. The subgroup (A2) with higher aromatase transcript level was associated with an increased seminal pH, a decreased sperm viability, low sperm percentage motility and low neutral α-glucosidase semen levels. Our data highlight the involvement of aromatase in motility and morphology of spermatozoa. Thus, this enzyme could bring new insights about quality and fertilising capacity of human spermatozoa.

Characterization of 3-hydroxyisobutyrate dehydrogenase, HIBADH, as a sperm-motility marker.

PURPOSE: Asthenozoospermia is a major cause of male infertility. However, the molecular mechanisms underlying sperm-motility defects remain largely unknown in the majority of cases. In our previous study, we applied a proteomic approach to identify unknown proteins that were downregulated in spermatozoa with low motility compared to spermatozoa with good motility. Several sperm motility- related proteins have been identified. In this study, 3-hydroxyisobutyrate dehydrogenase (HIBADH), one of the proteins identified using the proteomic tools, is further characterized. METHODS: Reverse-transcription polymerase chain reactions (RT-PCR), western blotting, and immunofluorescence assays (IFA) were preformed to investigate the expression pattern. The enzymatic activity of HIBADH was evaluated in sperm with good (>50 %), moderate (< 50 %) and lower motility (< 20 %). RESULTS: Using RT-PCR, we found that transcripts of HIBADH are enriched in the cerebellum, heart, skeletal muscle, uterus, placenta, and testes of male humans. In western blotting, it is expressed in the placenta, testes, and spermatozoa. During spermiogenesis, HIBADH is located at the mid-piece (a specialized development from the mitochondria) of elongating, elongated, and mature sperm. The enzymatic activity of HIBADH in sperm with moderate and lower motility were significantly reduced compared with good motility (P<0.0001 and P<0.05, respectively). CONCLUSIONS: Our study indicated that HIBADH is involved in the mitochondrial function of spermatozoa, and maintains sperm motility. It may serve as a sperm-motility marker.

RNASET2 in human spermatozoa and seminal plasma: a novel relevant indicator for asthenozoospermia.

Adequate sperm motility is requisite for human fertilization, whereas the underlying causes or mechanisms of impaired sperm motility, asthenozoospermia, still remain largely unknown. RNASET2 (Ribonuclease T2) may be one of the effectors modulating human sperm motility. We determined if there is a correlation between RNASET2 expression levels in human semen from asthenozoospermia and fertile individuals. Thus, RNASET2 expression levels in spermatozoa and seminal plasma of healthy and asthenozoospermia individuals were evaluated using Western blot, laser scanning confocal microscope analysis, ELISA and flow cytometry. The results revealed that RNASET2 expression was identified in both human spermatozoa and seminal plasma. In spermatozoa from fertile individuals, it was localized to the acrosome, neck and the middle piece of tail regions. However, in spermatozoa from asthenozoospermia individuals (n = 67), RNASET2 staining was especially more frequent and evident in the neck and middle piece than that in fertile individuals (n = 59, p < 0.01). Similarly, higher RNASET2 expression was also apparent in seminal plasma from asthenozoospermia than in fertile individuals (p < 0.01). Moreover, purified RNASET2 had an inhibitory effect on sperm motility, especially on progressive motility (n = 23, p < 0.05). In conclusion, higher expression of RNASET2 in the semen of asthenozoospermia individuals may contribute to sperm motility impairment.