Evaluation of LDH Activity and Its Relationship with Fructose Levels in Seminal Plasma of Normospermic and Asthenospermic Males

Abstract

Background and Objectives: sperm motility mainly depends on aerobic glycolysis, and lactate dehydrogenase (LDH) is a key enzyme in the last step of this process. On the other hand, fructose is considered as the main energy source for sperm motility. Therefore, this study aimed to evaluate the LDH activity and fructose levels and their correlation with sperm motion parameters in normospermic and asthenozospermic males.Methods: In this case-control study, 96 normospermic and 96 asthenozospermic males were enrolled. The semen samples were analyzed by computer assisted semen analysis to identify sperm motion parameters. Seminal plasma fructose levels were measured using Seliwanoff's test. LDH activity in the samples was evaluated using commercially available kits. The obtained data were analyzed by SPSS software (version 16).Results: The levels of LDH activity and fructose concentration were not different between the two groups. There was no correlation found between LDH activity and fructose concentration or sperm motility parameters. However, LDH was significantly correlated with sperm concentration in both groups.Conclusion: Although LDH activity and fructose levels seem to be influenced by sperm motility, other factors such as sperm concentration and total sperm count can also affect their seminal plasma levels. Thus, when the sperm concentrations are equal in study groups, the level of these factors are expected to be equal.Keywords: Lactate Dehydrogenase, Fructose, Sperm Motility, CASA. two groups based on the results of their spermiogram. The subjects were matched in terms of age to prevent the confounding role of age in interpretation of results. The inclusion criteria consisted of the following: Total volume >2 ml, sperm concentration > 20×10 6 sperm/ml, and total sperm count > 40×10 6 . The exclusion criteria consisted of the following: Leukocytospermia (more than 1×10 6 leukocytes/ml of ejaculation), the presence of blood, mucosa, and pus in samples, and the period of more than 2 hours from sampling until analysis. Evaluation of LDH Activity and Its Relationship with Fructose Levels in Seminal Plasma of Normospermic and Asthenospermic Males Sample selectionThe samples with normal morphology (more than 30% of whole sperms) and normal concentration (more than 20×10 6 sperm/ml) with motility rate problems (total motility < 50%) were considered as the case group, while the samples with normal morphology, normal concentration, and normal motility rate (total motility > 50%) were considered as the control group. Semen analysisThe semen samples were collected in sterile containers by masturbation after a sexual abstinence period of 3-7 days. Volume of the ejaculates was measured immediately and the samples were incubated at 37°C for 15-60 minutes (liquefaction process). The unliquefied samples were excluded from the study. Manual semen analysis and computer aided/assisted semen analysis

Lactate dehydrogenase C4 (LDH-C4) is essential for the sperm count and motility: A case-control study

“…zero). This study's findings are in agreement with those of Gerez de Burgos et al (1979), 14 Hashemi et al (2016), 6 and Velasco et al (1993) 19 .…”

“…5 The LDH-C4 can be determined in seminal plasma and spermatozoa due to the outward diffusion or leakage of the enzyme from sperm cell and spontaneous destruction of sperm cells. 4,6 It also plays an important role in maintaining glycolysis and ATP production in the flagellum during sperm capacitation, therefore essential for male fertility. 6,7 The final step of glycolysis process, which is essential for continued production of ATP, is the transformation of pyruvate to lactate accompanied via oxidation of NADH to NAD+ (Figure 1).…”

“…4,6 It also plays an important role in maintaining glycolysis and ATP production in the flagellum during sperm capacitation, therefore essential for male fertility. 6,7 The final step of glycolysis process, which is essential for continued production of ATP, is the transformation of pyruvate to lactate accompanied via oxidation of NADH to NAD+ (Figure 1). This reaction is stimulated by LDH-C4.…”

Machine Learning Approach for Muscovy Duck (Cairina moschata) Semen Quality Assessment

“…Each diluted sperm sample was further divided into three equal parts-seminal plasma (sp), water sperm extract (w), and Triton extract (t). Both extracts and seminal plasma were used to analyse the enzyme activity of alkaline phosphatase (AP) [30,31], creatine kinase (CK) [27,36], lactate dehydrogenase (LDH) [29], and γ-glutamyl-transferase (GGT) [24] by biochemical semi-automatic analyser BA-88A (Mindray, China), following Via Campania kits (Italy) manufacturer instructions: for GGT-Tris buffer 100 mM pH 8.25,glycil-glycine 100 mM, L-Glutamyl-4-nitroanilide 4 mM, wavelength 405 nm; for AP-DEA buffer pH 9.8 1 M, MgCl2 0.5 mM, 4-Nitrophenilphosphate 10 mM, wavelength 405 nm; for CK-Imidazole buffer 29 mM pH 6.50, creatine phosphate 30 mM, glucose 20 mM, N-Acetyl-L-cysteine 20 mM, magnesium acetate 10 mM, EDTA 2 mM, ADP 2 mM, NADP 2 mM, AMP 5 mM, Di(adenosine-5) pentaphosphate 12 MikroM, glucose-6-phosphate-dehydrogenase > 3 kU/L, hexokinase > 3 kU/L, wavelength 340 nm; for LDH-phosphate buffer pH7.50 50 mM, sodium pyruvate 0.60 mM, NADH 0.18 mM, wave length 340 nm. Distilled water was used as an assay standard, which was analysed in the system before measuring the samples.…”

“…An additional functional criterion encompasses specific seminal biochemical parameters, such as sperm-produced enzymes such as lactate dehydrogenase (LDH) and creatine phosphokinase (CK), which can reflect sperm maturity, mitochondrial activity, and apoptosis pathways [26,27]. LDH activity has also been directly associated with sperm motility [28,29]. Semen contains low levels of the enzymes CK and LDH, which play important roles in energy metabolism, but whose levels can rise in response to cellular damage or death.…”