This study was performed to determine the effects of nitric oxide on human sperm cell function. Semen samples were obtained from normal healthy volunteers. Motile spermatozoas collected by swim-up method were incubated up to 24 hours in Ham's F-10 medium supplemented with a various concentration of sodium nitroprusside (nitric oxide releasing agent). Sperm motility, hyperactivation, acrosome reaction rate, and acrosin activity were determined. The results are as follows; 1. 1mM of SNP resulted in a significant decrease in sperm motility (44.8%+/- 8.9%:78.1%+/-6.3%, and hyperactivation (10.4%+/-6.4%:477%+/-9.5%) after incubation for 3 hours compared with the control group (Ham's F-10 alone), but had no effect on acrosome reaction. 2. At 100muM SNP, sperm motility was reduced after incubation for 6 hours (54.8%+/- 3.2%) compared with that of the control group (82.7% +/- 8.9%), but hyperactivation and acrosome reaction were not affected. 3. However, a lower concentration (less than 101M) of SNP had no effect on sperm motility and hyperactivation for 8 hours of incubation but significantly decreased them when incubation periods were increased up to 24 hours compared with the control group. On the other hand, 1muM and l0muM SNP significantly increased the acrosome reaction rate in both acrosomal status (17.3%+/-5.2%,23.5%+/-4.7%, respectively) and acrosin activity (34.3muIU+/- 10.5muIU, 45.6muIU+/-5.6muIU, respectively) as compared with the control group (7.0%+/-4.0%, 9.5muIU+/-3.4muIU). These results indicate that SNP, NO releasing agent, has a dose-dependent effects on the sperm cell function. Therefore it may positively affect the fertilization by promoting acrosomal reaction at a lower concentration (less than 101M).
Acrosin ; Acrosome Reaction ; Fertilization ; Hand ; Healthy Volunteers ; Humans* ; Nitric Oxide* ; Nitroprusside ; Semen ; Sperm Motility ; Spermatozoa*

OBJECTIVETo study the effect of Percoll selection technique on normal morphology and acrosin activity of human spermatoza.

METHODSThe sperm morphology and sperm acrosin activity were analyzed by automated sperm morphology analyzer(ASMA) and spectrocolorimetry.

RESULTSThe normal morphology sperm rate and acrosin activity were significantly increased after Percoll selection technique (P < 0.001).

CONCLUSIONPercoll selection technique could affect normal morphology sperm ratio and acrosin activity.

Acrosin ; metabolism ; Adult ; Centrifugation, Density Gradient ; Fertilization in Vitro ; Humans ; Male ; Spermatozoa ; cytology ; enzymology

OBJECTIVESTo investigate the effect of gamma-aminobutyric acid (GABA) on the sperm acrosin activity in normal men and positive antisperm antibody (AsAb) men.

METHODSSperm acrosin activity was detected by BAEE/ADH method.

RESULTSGABA could increase the sperm acrosin activity in normal and AsAb positive patients (P < 0.01). The results also indicated that GABA significantly increased Na(+)-K(+)-ATPase activity (P < 0.01), Ca(2+)-ATPase activity (P < 0.05) and SOD activity (P < 0.01) of sperm.

CONCLUSIONSThe results demonstrated that GABA could influence the sperm acrosin activity.

Acrosin ; metabolism ; Humans ; Male ; Spermatozoa ; drug effects ; immunology ; metabolism ; gamma-Aminobutyric Acid ; pharmacology

OBJECTIVESTo study the inhibitory effect of KF-950 on human acrosin and sperm acrosome.

METHODSHuman acrosin was extracted and purified with 2% acetic acid, and its residual activity was evaluated by BAEE/ADH assay after treated with different concentrations of KF-950. ABC assay was used to observe the effect of KF-950 on human acrosome with Biotin-PSA as a probe.

RESULTS1. The activity of normal sperm acrosin was (37.65 +/- 4.47) U/L. 2. The residual activity was inversely related to the concentration of KF-950 (r = -0.998), and had a dose-response curve. The result could be described by Y = 7.57-1.895X. 3. With increase of KF-950 concentration and prolongation of action time, the staining rate of acrosome obviously dropped (P < 0.01).

CONCLUSIONSKF-950 directly inhibits acrosin activity and assumely injures sperm acrosome. It might be a new kind of highly effective inhibitor.

Acrosin ; antagonists & inhibitors ; Acrosome ; metabolism ; Humans ; Male ; Spermatozoa ; drug effects ; physiology

OBJECTIVETo evaluate the inhibitory effect of Nandeshi, an acrosin inhibitor, on human acrosin activity.

METHODSWe collected sperm samples from 10 healthy fertile men and cultured them with Nandeshi at 30 degrees C for 5 minutes at the concentrations of 0. 100, 0.120, 0.144, 0.173, 0.207, 0.249, 0.299, 0.358 and 0.430 mmol/L, with the controls treated with a well-known acrosin inhibitor N-alpha-p-tosyl-L-lysine chloromethylketone (TLCK) at 150.0, 189.8, 213.6, 240.3, 270.3, 304.1 and 342.1 mmol/L. Then we determined the residual activity of human acrosin by improved Kennedy assay.

RESULTSThe residual activity of acrosin was negatively correlated with the Nandeshi concentration, and Nandeshi exhibited an inhibition rate about 800 times that of TLCK.

CONCLUSIONNandeshi has a powerful inhibitory effect on human acrosin, and improved Kennedy assay is a simple, practical and highly sensitive technique for the detection of human acrosin activity.

Acrosin ; antagonists & inhibitors ; metabolism ; Contraceptive Agents, Female ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Humans ; Male ; Spermatozoa ; drug effects ; Tosyllysine Chloromethyl Ketone ; pharmacology

OBJECTIVESTo study the effect of antisperm antibody(AsAb) on human sperm acrosin activity.

METHODSAsAb and sperm acrosin activity were measured and analyzed in 3,432 infertile men and 65 fertile volunteers.

RESULTSAsAb positive rate was 10.20% in 3,432 case of male infertility, and 9.37% in 2,882 infertile males who received tests of sperm acrosin activity. Acrosin activity of infertility cases were lower than those of fertile cases(P < 0.001). The comparison between AsAb positive group and AsAb negative group infertility cases showed no significant differences of acrosin activity (P > 0.05). Between normal acrosin activity group and abnormal acrosin activity group, there was no significant difference of AsAb positive rate (P > 0.05).

CONCLUSIONSAntisperm antibody could not affect acrosin activity.

Acrosin ; metabolism ; Adult ; Autoantibodies ; analysis ; Case-Control Studies ; Humans ; Infertility, Male ; immunology ; Male ; Semen ; chemistry ; Spermatozoa ; enzymology ; immunology

AIMTo develop a method for assessing sperm function by measuring released acrosin activity during the acrosome reaction (AR).

METHODSHuman semen samples were obtained from 24 healthy donors with proven fertility after 3-7 days of sexual abstinence. After collection, samples were liquefied for 30 min at room temperature. Standard semen parameters were evaluated according to World Health Organization (WHO) criteria. Calcium ionophore A23187 and progesterone (P4) were used to stimulate the sperm to undergo AR. After treatment, sperm were incubated with the supravital dye Hoechst33258, fixed in a glutaraldehyde-phosphate-buffered saline solution, and the acrosomal status was determined by fluorescence microscopy with fluorescein isothiocyanate-labeled Pisum sativum agglutinin (FITC-PSA). The percentage of sperm undergoing AR (AR%) was compared to sperm acrosin activities as assessed by spectrocolorimetry. The correlation between AR% and acrosin activity was determined by statistical analysis.

RESULTSThe AR% and released acrosin activity were both markedly increased with A23187 and P4 stimulation. Sperm motility and viability were significantly higher after stimulation with P4 versus stimulation with A23187 (P < 0.001). There was a significant positive correlation between released acrosin activity and AR% determined by FITC-PSA staining (r=0.916, P < 0.001).

CONCLUSIONSpectrocolorimetric measurement of released acrosin activity might serve as a reasonable alternative method to evaluate AR.

Acrosin ; physiology ; Acrosome Reaction ; Adult ; China ; Humans ; Male ; Progesterone ; pharmacology ; Semen ; drug effects ; physiology ; Sperm Motility ; drug effects ; physiology

OBJECTIVETo investigate the effect of sperm acrosin activity on the IVF-ET outcome.

METHODSWe analyzed sperm parameters, morphology and acrosin activity for 909 infertile husbands by computer-assisted self-assessment (CASA), modified Papanicolaou staining and N-alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA), respectively, and detected the rates of fertilization, cleavage, quality embryos, embryo cryopreservation, implantation, clinical pregnancy and abortion. The wives were identified as normal or with mere oviduct problems.

RESULTSThe rate of normal sperm morphology and sperm motility, vitality, rapid progressive velocity and concentration were significantly lower in the abnormal acrosin activity group than in the normal one (P < 0.01). Significant positive correlations were observed between acrosin activity and the above-mentioned semen parameters (P < 0.01). There were no significant differences in the number of retrieved eggs, the rates of cleavage, quality embryos, embryo cryopreservation, non-embryo transfer cycles and miscarriages, and the number of transferred embryos between the two groups (P > 0.05). The fertilization rate, the percentage of transfer cycles with only 1 embryo and the rate of implantation and clinical pregnancy were notably higher in the normal acrosin activity group than in the abnormal one (P < 0.01).

CONCLUSIONSperm acrosin activity is closely related with semen parameters, and it helps to predict the sperm fertilizing capacity and IVF-ET outcome.

Acrosin ; metabolism ; Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Male ; Male ; Pregnancy ; Pregnancy Rate ; Semen Analysis ; Spermatozoa ; enzymology

OBJECTIVETo analyze the activity of human sperm acrosin and semen parameters in male infertile patients and discuss the effect of sperm acrosin activity on semen quality.

METHODSActivity of human sperm acrosin, PMN-elastase, fructose, alpha-glucosidase, zinc, acid phosphatase levels and HOST of 214 male infertile patients were detected. Semen analysis was performed using WLJY-9000 WeiLi colorful semen quality analyzer. There were 111 cases of normal activity of human sperm acrosin (48.2 - 218.7 microIU/10(6) sperm), 103 cases abnormal (< 48.2 microIU/10(6) sperm). Using the group of normal activity of sperm acrosin to be the control, semen parameters was analyzed and compared with those of the group of abnormal activity of sperm acrosin.

RESULTSThere were significant difference (P < 0.001) between the 2 groups (normal and abnormal) in the areas of sperm density, motile sperm rate, percentage of grade (a + b) sperm and HOST. There were also significant difference in PMN-elastase, fructose and alpha-glucosidase (P < 0.05). There was no difference among sperm volume, zinc and acid phosphatase (P > 0.05).

CONCLUSIONThere was a strong correlation between the activity of human sperm sperm acrosin and semen quality. Activity of sperm acrosin is a reliable index of semen quality.

Acrosin ; metabolism ; Adult ; Humans ; Infertility, Male ; enzymology ; physiopathology ; Male ; Middle Aged ; Semen ; chemistry ; Sperm Count ; Sperm Motility ; Spermatozoa ; enzymology

Acrosin ; immunology ; isolation & purification ; Humans ; Male ; Membrane Proteins ; immunology ; isolation & purification ; Protein Binding ; Spermatozoa ; metabolism ; Zona Pellucida ; metabolism