Aim: To find out whether the response of testicular oxidative stress parameters to hexachlorocyclohexane (HCH) is species specific. Methods: In rats and mice (n=5 in each group), HCH was administered at a dose of 20 m...Aim: To find out whether the response of testicular oxidative stress parameters to hexachlorocyclohexane (HCH) is species specific. Methods: In rats and mice (n=5 in each group), HCH was administered at a dose of 20 mg/kg/day intraperitoneally for 30 days in 0.1 ml of refined groundnut oil. The control groups received equal volume of the vehicle. Animals were sacrificed 24 hours after the last injection and various oxidative stress parameters were measured immediately. Results: The level of both endogenous as well as FeSO_4 and ascorbic acid-stimulated lipid peroxidation was increased significantly in the HCH-treated rats, whereas the pattern was just the reverse in case of mice. Although the level of H_2O_2 content increased in response to HCH in both groups, a totally different trend was observed for the activity of the principal H_2O_2-metabolising enzyme, catalase. In case of rats, a significant decline in the activity of catalase was recorded in response to HCH whereas a sharp augmentation in the enzyme activity was noticed in mice. Similarly, the decreased activity of superoxide dismutase observed in rats remained unaltered in mice. Conclusion: HCH induces oxidative stress in the testis of both rats and mice. However, the pattern of response of testicular oxidative stress parameters seems to be species specific.展开更多
To study the major differences in the distribution of spermatozoa proteins in infertile men with varicocele by comparative proteomics and validation of their level of expression. The study-specific estimates for each ...To study the major differences in the distribution of spermatozoa proteins in infertile men with varicocele by comparative proteomics and validation of their level of expression. The study-specific estimates for each varicocele outcome were combined to identify the proteins involved in varicocele-associated infertility in men irrespective of stage and laterality of their clinical varicocele. Expression levels of 5 key proteins (PKARIA, AK7, CCT6B, HSPA2, and ODF2) involved in stress response and sperm function including molecular chaperones were validated by Western blotting. Ninety-nine proteins were differentially expressed in the varicocele Eroup. Over 87% of the DEP involved in major energy metabolism and key sperm functions were underexpressed in the varicocele group. Key protein functions affected in the varicocele group were spermatogenesis, sperm motility, and mitochondrial dysfunction, which were further validated by Western blotting, corroborating the proteomics analysis. Varicocele is essentially a state of energy deprivation, hypoxia, and hyperthermia due to impaired blood supply, which is corroborated by down-regulation of lipid metabolism, mitochondrial electron transport chain, and Krebs cycle enzymes. To corroborate the proteomic analysis, expression of the 5 identified proteins of interest was validated by Western blotting. This study contributes toward establishing a biomarker "fingerprint" to assess sperm quality on the basis of molecular parameters.展开更多
Dysfunctional sperm maturation is the primary reason for the poor sperm motility and morphology in infertile men.Spermatozoa from infertile men were fractioned on three-layer density gradient(80%,60%,and 40%).Fraction...Dysfunctional sperm maturation is the primary reason for the poor sperm motility and morphology in infertile men.Spermatozoa from infertile men were fractioned on three-layer density gradient(80%,60%,and 40%).Fraction 1(Fl)refers to the least mature stage having the lowest den sity,whereas the fraction 4(F4)in eludes the most dense and morphologically mature motile spermatozoa.Fraction 2(F2)and fraction 3(F3)represent the intermediate stages.Proteins were extracted and separated by dimensional gel.Bands were digested with trypsin and analyzed on a LTQ-Orbitrap Elite hybrid mass spectrometer system.Functional annotations of proteins were obtained using bioinformatics tools and pathway databases.A total of 1585 proteins were detected in the four fractions of spermatozoa.A dysregulated protein turnover and protein folding may lead to accumulation of defective proteins or proteins that otherwise would have been eliminated during the process of maturation,resulting in the impairment of sperm fun ction.Aberra nt chaper one expressi on may be a major con tributing factor to the defective sperm function.Androge n receptor was predicted as a transcription regulator in one of the networks and the affected pathways were chaperone-mediated stress response,proteosomal pathway,and sperm function.The down regulation of key pathways and protei ns which compromises the fertilizi ng potential of spermatozoa may provide in sight into the mecha nisms that lead to male infertility.展开更多
文摘Aim: To find out whether the response of testicular oxidative stress parameters to hexachlorocyclohexane (HCH) is species specific. Methods: In rats and mice (n=5 in each group), HCH was administered at a dose of 20 mg/kg/day intraperitoneally for 30 days in 0.1 ml of refined groundnut oil. The control groups received equal volume of the vehicle. Animals were sacrificed 24 hours after the last injection and various oxidative stress parameters were measured immediately. Results: The level of both endogenous as well as FeSO_4 and ascorbic acid-stimulated lipid peroxidation was increased significantly in the HCH-treated rats, whereas the pattern was just the reverse in case of mice. Although the level of H_2O_2 content increased in response to HCH in both groups, a totally different trend was observed for the activity of the principal H_2O_2-metabolising enzyme, catalase. In case of rats, a significant decline in the activity of catalase was recorded in response to HCH whereas a sharp augmentation in the enzyme activity was noticed in mice. Similarly, the decreased activity of superoxide dismutase observed in rats remained unaltered in mice. Conclusion: HCH induces oxidative stress in the testis of both rats and mice. However, the pattern of response of testicular oxidative stress parameters seems to be species specific.
文摘To study the major differences in the distribution of spermatozoa proteins in infertile men with varicocele by comparative proteomics and validation of their level of expression. The study-specific estimates for each varicocele outcome were combined to identify the proteins involved in varicocele-associated infertility in men irrespective of stage and laterality of their clinical varicocele. Expression levels of 5 key proteins (PKARIA, AK7, CCT6B, HSPA2, and ODF2) involved in stress response and sperm function including molecular chaperones were validated by Western blotting. Ninety-nine proteins were differentially expressed in the varicocele Eroup. Over 87% of the DEP involved in major energy metabolism and key sperm functions were underexpressed in the varicocele group. Key protein functions affected in the varicocele group were spermatogenesis, sperm motility, and mitochondrial dysfunction, which were further validated by Western blotting, corroborating the proteomics analysis. Varicocele is essentially a state of energy deprivation, hypoxia, and hyperthermia due to impaired blood supply, which is corroborated by down-regulation of lipid metabolism, mitochondrial electron transport chain, and Krebs cycle enzymes. To corroborate the proteomic analysis, expression of the 5 identified proteins of interest was validated by Western blotting. This study contributes toward establishing a biomarker "fingerprint" to assess sperm quality on the basis of molecular parameters.
文摘Dysfunctional sperm maturation is the primary reason for the poor sperm motility and morphology in infertile men.Spermatozoa from infertile men were fractioned on three-layer density gradient(80%,60%,and 40%).Fraction 1(Fl)refers to the least mature stage having the lowest den sity,whereas the fraction 4(F4)in eludes the most dense and morphologically mature motile spermatozoa.Fraction 2(F2)and fraction 3(F3)represent the intermediate stages.Proteins were extracted and separated by dimensional gel.Bands were digested with trypsin and analyzed on a LTQ-Orbitrap Elite hybrid mass spectrometer system.Functional annotations of proteins were obtained using bioinformatics tools and pathway databases.A total of 1585 proteins were detected in the four fractions of spermatozoa.A dysregulated protein turnover and protein folding may lead to accumulation of defective proteins or proteins that otherwise would have been eliminated during the process of maturation,resulting in the impairment of sperm fun ction.Aberra nt chaper one expressi on may be a major con tributing factor to the defective sperm function.Androge n receptor was predicted as a transcription regulator in one of the networks and the affected pathways were chaperone-mediated stress response,proteosomal pathway,and sperm function.The down regulation of key pathways and protei ns which compromises the fertilizi ng potential of spermatozoa may provide in sight into the mecha nisms that lead to male infertility.