Oxidative stress (OS) and inflammation are common causes of male infertility; antioxidants in spermatozoa and especially in seminal plasma play a protective role. The study aims to evaluate whether OS lipid mediators (F-2-isoprostanes: F-2-IsoPs; Resolvin D1: RvD1) measured in seminal plasma, and Trolox equivalent antioxidant capacity (TEAC), measured in both seminal plasma and spermatozoa, are associated with male infertility. Semen samples (18 fertile and 62 infertile subjects) were examined following WHO guidelines and with a mathematically elaborated transmission electron microscopy analysis (fertility index-FI-, % sperm pathologies). F-2-Isops were measured by GC/NICI-MS/MS, RvD1 by ELISA, and TEAC by a commercially available antioxidant assay. F-2-Isops, RvD1, and sperm TEAC (positively interrelated with each other) correlated negatively with seminal parameters and FI and positively with sperm pathologies. F-2-Isops, RvD1, and sperm TEAC levels were significantly higher in infertile vs fertile subjects (p < 0.001). ROC curve analysis demonstrated that sperm TEAC (J index: 0.13 mM) and seminal RvD1 (J index: 38.26 pg/mL) discriminated between fertile and infertile subjects. Spermatozoa stimulate antioxidant capacity in the presence of an OS environment; this data suggests an association in which antioxidant defences may vary in relation to the surrounding seminal plasma. Moreover, sperm TEAC, and to a lesser extent seminal RvD1, emerge as potential markers for identifying infertile patients.

BackgroundThe beneficial effects of balneotherapy in osteoarthritis (OA) using the sulfate-arsenicalferruginous Levico water (LW) have been recognized by several clinical studies. However, the specific biological properties of LW are partially understood.ObjectiveThe present study aimed at evaluating the ability of LW in the regulation of apoptosis, oxidative stress and of the gene expression of a panel of articular cartilage markers and MicroRNA (miRNA) in human osteoarthritic chondrocytes exposed to interleukin (IL)-1β.MethodsChondrocytes were obtained from the femoral heads of patients with OA who underwent surgery for total hip prostheses. The cells were incubated for 24 h and 48 h with different concentrations of LW, alone or in combination with IL-1β (10 ng/ml). Apoptosis and mitochondrial superoxide anion production were detected by cytometry. Gene levels of antioxidant enzymes, B-cell lymphoma (BCL)2, cytokines [IL-1β, IL-6, tumor necrosis factor (TNF)-α], metalloproteinases (MMPs)1 and 13, type II collagen (Col2a1), aggrecan, and of a pattern of miRNA were evaluated by Quantitative Real-Time PCR analysis (PCR). The involvement of nuclear factor (NF)-κB was investigated by PCR and immunofluorescence and by pre-incubation with a specific inhibitor (BAY 117082, IKKα/β).ResultsLW at 50% or 25% concentration counteracted the negative effects of IL-1β on cell viability, apoptosis and mitochondrial superoxide anion production and on the gene expression of antioxidant enzymes, in a dose-dependent manner. The treatment of chondrocytes with 50% of LW significantly inhibited the gene expression of pro-inflammatory cytokines, MMPs and of miR-34a and miR-181a, while it upregulated miR-140 in IL-1β stimulated cells. The pre-incubation with BAY 11–7082, IKKα/β reduced the damage induced by IL-1β, and, interestingly, potentiated the properties of LW.ConclusionsOur data demonstrated the ability of LW to regulate apoptosis, oxidative stress and gene expression of main factors implicated in OA pathogenesis through a possible modulation of NF-κB signaling pathway. This study supports the use of balneotherapy with a sulfate-arsenical-ferruginous mineral water in the treatment of OA.

Seminal plasma regulates sperm function; oxidative stress (OS) and inflammation can impair semen biochemical homeostasis. This study investigated seminal biochemical/enzymatic compounds and Trolox Equivalent Antioxidant Capacity (TEAC) in seminal plasma and spermatozoa from normozoospermic men (>5th percentile) assessing their relationship with sperm quality. The strongest significant (p < 0.001 ***) negative correlations (r) were found for normal sperm morphology and albumin (ALB, r = -0.515 ***), cholesterol (CHO, r = -0.386 ***), lactate dehydrogenase (LDH, r = -0.514 ***), sperm TEAC (r = -0.438 ***) and homocysteine (Hcy; r = -0.381 ***); sperm progressive motility with ALB (r = -0.495 ***), CHO (r = -0.349 ***), LDH (r = -0.355 ***) and sperm TEAC (r = -0.295 ***); sperm vitality with creatine kinase (CK, r = -0.486 ***) and LDH (r = -0.380 ***). Total prostate-specific antigen (TPSA) was positively correlated with sperm quality. After stratification, subjects with low-normal sperm parameters showed significantly higher levels of ALB, CHO, CK, LDH, interleukin 6 (IL-6), Hcy and sperm TEAC vs. high-normal parameter subjects. ALB, Hcy and LDH were increased in individuals with progressive motility, normal morphology and vitality < 25th percentile indicating their involvement in OS, inflammation and accessory gland activity. These findings suggest that World Health Organization lower reference limits are not a strict cutoff between normal and abnormal semen quality but rather define a baseline of biochemical efficiency and sperm function, highlighting the synergy between spermatozoa and seminal plasma.

Oxidative stress (OS) is a major cause of defective sperm function. During laboratory handling, gametes are exposed to OS, potentially mitigated by in vitro antioxidant supplementation. This study evaluates the protective role of caffeic acid (CAF) on basal human semen and under induced OS. First, six semen samples from normozoospermic donors were incubated with CAF concentrations ranging from 50 to 500 µM at 37 °C for 2 h. Sperm motility and DNA integrity (acridine orange) were evaluated. Then, ten semen samples were divided into four aliquots and incubated, respectively, with CAF at 100 µM, H2O2 at 2 mM, or H2O2 at 2 mM + CAF at 100 µM, or untreated. Motility, DNA integrity, acrosome status (Pisum sativum agglutinin), OS quantified by F2-isoprostanes (ELISA), and expression of Nrf2, Keap1, and HO-1 (qRT-PCR) were assessed. CAF at 100 µM improved progressive motility without damaging DNA and was selected for subsequent experiments. CAF showed protective effects on sperm damage induced by H2O2 treatment, restoring motility, DNA integrity, and acrosome status and reducing F2-isoprostane levels. Nrf2 and HO-1 expression were upregulated by CAF, downregulated by H2O2, and restored by the co-treatment. CAF supplementation may protect human spermatozoa during in vitro handling by reducing OS, improving several sperm parameters, with a possible mechanism of action involving the Nrf2 pathway.

Despite the recognized role of oxidative stress (OS) in sperm function, limited data exist on Nuclear factor-E2-related factor 2 (Nrf2) pathway modulation in relation to reliable oxidative damage markers in human semen. In this study, 79 semen samples were collected from men undergoing semen analysis and grouped as varicocele (V, no. 22), urogenital infections (UI, no. 23), unknown fertility status without pathologies (UFS, no. 15), and fertile controls (F, no. 19). After semen analysis, ELISA were used to quantify F-2-Isoprostane (F-2-IsoPs) level, a marker of lipid peroxidation, and Nrf2 in seminal plasma and spermatozoa. The Nrf2 pathway (Keap1, Nrf2, Bach1, HO-1) was assessed in spermatozoa by qRT-PCR. Seminal plasma and sperm Nrf2 positively correlated with F-2-IsoPs (p < 0.001) and negatively with sperm vitality. Sperm Nrf2 also inversely correlated with progressive motility (p < 0.05). Seminal F-2-IsoP levels were lower in F than in the other groups. Sperm Nrf2 was significantly lower in F versus V and UI (p < 0.001) and UFS (p < 0.05), while seminal plasma Nrf2 levels did not differ among groups. qRT-PCR suggested Nrf2 pathway activation mainly in V and UI, consistent with increased OS. Elevated F-2-IsoPs, a marker of poor sperm quality, and sperm Nrf2 could suggest OS-driven Nrf2 activation, providing complementary biomarkers of oxidative status in male reproductive health.

Omega-3 polyunsaturated fatty acids (n-3 PUFAs) are recognized for their beneficial effects on male fertility. This study evaluated the protective effects of dietary n-3 PUFAs from extruded linseed, alone or combined with the alga Padina pavonica, against in vitro lipopolysaccharide (LPS)-induced sperm dysfunction in rabbits. Twelve bucks were fed for 60 days a control diet (CNT), a diet containing 5% extruded linseed (L), or 5% extruded linseed plus 0.2% P. pavonica extract (LPP). Ejaculates were exposed in vitro to increasing LPS concentrations (0, 400, and 600 µg/mL), and sperm motility was evaluated at 0, 1, 2, and 4 h using computer-assisted sperm analysis. LPS markedly impaired sperm motility in the CNT group, increasing the percentage of static spermatozoa (p < 0.001) and reducing sperm progressive motility (p < 0.001), with complete immobility observed at 600 µg/mL after 4 h. Conversely, sperm from L and LPP groups maintained significantly higher progressive motility, lower static sperm, and improved kinematic parameters throughout the LPS challenge (p < 0.05). Dietary n-3 PUFA supplementation also attenuated LPS-induced TLR4 activation and reduced lipid peroxidation, as indicated by lower seminal TBARS levels. No histological alterations were detected in the male reproductive tract. These findings indicate that n-3 PUFA supplementation, particularly linseed combined with algae, mitigates LPS-induced sperm dysfunction in vitro.

F4-Neuroprostanes (F4-NeuroPs), oxidative metabolites of docosahexaenoic acid, act as bioactive lipid mediators enhancing sperm motility and induce capacitation-like changes in vitro. Their biological action is proposed to involve sperm ion channels, in particular ryanodine receptors (RyRs), which regulate intracellular calcium homeostasis. We evaluated the effects of dantrolene, a RyR inhibitor, on motility and vitality of a selected spermatozoa at different concentrations (10, 30, 50, 100 μM). Then sperm motility, acrosome integrity, and RyR localization following co-incubation with dantrolene (D50 or D100 μM) and 4-/10-F4t-NeuroPs (7 ng) were investigated. Acrosomal status was assessed using Pisum sativum agglutinin (PSA) staining and RyR localization by immunofluorescence. D50 was identified as the minimum effective dose to induce significant reductions in sperm motility. F4-NeuroPs significantly increased rapid progressive motility versus controls. Co-incubation with F4-NeuroPs + D50 reduced rapid motility and increased in situ and circular movement. The acrosome staining appeared altered or absent to different percentages, and RyR localization was also seen in the midpiece. These findings suggested that F4-NeuroPs enhance sperm motility via RyR-mediated pathways, as confirmed by dantrolene inhibition. Accordingly, our results underscore the physiological relevance of RyRs in sperm function and suggest new insights into lipid-based mechanisms regulating sperm motility.

Lipopolysaccharide (LPS)-induced inflammation impairs sperm function; however, its impact on ejaculated rabbit sperm remains unexplored. This dose-response study aims to determine the LPS concentration that negatively affects sperm motility in vitro, while also providing the first identification of TLR4 localization on rabbit spermatozoa. Additionally, it evaluates malondialdehyde (MDA) levels in seminal plasma as an indicator of oxidative stress. Sperm motility was analyzed using computer-assisted sperm analysis (CASA) after incubation with increasing LPS concentrations (0, 50, 100, 200, 400, 600, and 800 µg/mL) at multiple time points (0, 1, 2, and 4 h). LPS doses ≥ 400 µg/mL significantly reduced progressive and non-progressive motility, as well as curvilinear velocity (all p < 0.001), while increasing the proportion of static spermatozoa (p < 0.05). Receiver operating characteristic (ROC) analysis identified 300 µg/mL as the threshold dose for motility decline. Immunofluorescence revealed TLR4 localization in the midpiece of sperm tails, with weak labeling in control samples and a marked increase after 4 h of incubation with 400 μg/mL LPS. MDA levels were assessed using the thiobarbituric acid reactive substances (TBARS) assay with a colorimetric kit, showing no significant effect of LPS treatment. No correlation was found between MDA and other semen parameters. ccThese findings identify TLR4 on rabbit sperm for the first time and establish a threshold LPS dose for future in vitro studies.

Many male reproductive pathologies and a part of undiagnosed infertility share an oxidative stress (OS) etiology with high reactive oxygen species and cytokine concentrations. The lack of reliable biomarkers to quantify oxidative injury is a crucial problem in the field of male infertility. In this observational study, IL-1β and the OS markers malondialdehyde (MDA) and F2-isoprostanes (F2-IsoPs) were quantified in seminal plasma of 46 infertile patients with varicocele, genitourinary infections, idiopathic infertility, and 11 fertile men. Semen analysis was performed following WHO guidelines, IL-1β was determined by ELISA, MDA was quantified by HPLC, and F2-IsoPs by GC/NICI-MS analysis. F2-IsoPs were immunolocalized in spermatozoa of fertile and infertile subjects. Results indicated that F2-IsoP, MDA, and IL-1β seminal levels positively correlated pairwise (p < 0.001) and showed negative correlations with sperm parameters (p < 0.001). Then, the studied population was grouped following the cause of infertility and the variables were compared between the different groups and a control sample. Seminal IL-1β, F2-IsoPs, and MDA were significantly higher in varicocele (p < 0.001, for MDA p < 0.01) and genitourinary infections (p < 0.001, for IL-1β p < 0.01) groups than those observed in fertile subjects. F2-IsoPs seemed to discriminate more accurately than MDA the different conditions, in particular idiopathic infertility. ROC curves demonstrated that the three analyzed indices were able to discriminate fertile and infertile patients. The immunofluorescence studies showed a low presence of F2-IsoPs in spermatozoa of fertile men and an evident labeling in the tail, and cytoplasmic residues of spermatozoa from infertile patients. In conclusion, this data confirmed that F2-IsoP level is a suitable marker of OS in seminal plasma, even more accurate than MDA and can be proposed for measuring OS in the clinical setting.

30 percent of infertile men are diagnosed with idiopathic infertility. This study aimed to assess oxidative stress in the semen of 77 patients with idiopathic infertility by measuring F2-isoprostane (F2-IsoP), resolvin D1 (RvD1) levels, and semen parameters. The presence and localization of 8-IsoProstaglandin F2α were determined using immunofluorescence. No significant correlations were observed for F2-IsoP and RvD1 levels with the semen variables. Based on F2-IsoP levels, individuals were classified into two groups: Group 1 (F2-IsoPs ≤ 29.96 ng/mL, 51%) and Group 2 (F2-IsoPs > 29.96 ng/mL, 49%). In comparison to Group 1, Group 2 showed significantly higher F2-IsoP levels (13.33 ng/mL vs. 44.80 ng/mL; p < 0.05), a lower progressive motility percentage (30% vs. 25%; p < 0.05), and increased RvD1 levels (36.09% vs. 44.94%). Immunofluorescence analysis revealed a different localization of 8-IsoProstaglandin F2α in the ejaculated sperm of Group 1 compared to that observed in Group 2. A weak signal was detected in the sperm tail (Group 1, 79.1% vs. Group 2, 36.9; p < 0.01). In spermatozoa of Group 2 patients, a strong signal in the acrosome, midpiece, and tail was highlighted. These findings suggest the need to test oxidative stress during routine semen analysis in patients with idiopathic infertility to improve diagnosis and treatment.

Growing evidence demonstrates the critical roles of long non-coding RNAs (lncRNAs) in osteoarthritis (OA) pathogenesis. The lncRNA XIST is one of the most commonly studied; however, its function remains unclear. This study aimed to research the molecular mechanism of XIST in human OA chondrocytes. Cells were transfected with small interfering RNA against XIST or with a microRNA (miR)-146a inhibitor in the presence of interleukin (IL)-1β. Viability was detected using MTT; apoptosis using cytometry; and XIST, miR-146a, B-cell lymphoma (BCL)2, and metalloproteinase (MMP)-13 expression using real-time PCR. The analysis of p50 and p65 nuclear factor (NF)-κB was conducted using PCR and immunofluorescence. Our findings showed that XIST was highly expressed in OA chondrocytes when compared to T/C-28a2 lines. Furthermore, XIST silencing significantly promoted survival and limited apoptosis, with a concomitant over expression of BCL2, reduction in MMP-13 mRNA, and NF-κB activation after IL-1β stimulus. Conversely, miR-146a was significantly down-regulated in OA cells, while its levels were increased following XIST silencing; moreover, miR-146a inhibition induced opposite results to those caused by XIST. Finally, the down-regulation of XIST was correlated to the over-expression of miR-146a, with a consequent modulation of BCL2, MMP-13, and NF-κB. This study suggests an influence of the XIST/miR-146a axis on the viability, apoptosis, and matrix degradation occurring in OA.

This study evaluated the effects of chlorogenic acid (CGA) on human semen and on oxidative stress (OS) induced in vitro in human spermatozoa. After the treatment of the basal semen with 100 µM CGA, rapid and slow sperm progressive motility were evaluated and seminal F2-Isoprostanes (F2-IsoPs), a marker of OS, were quantified by ELISA. In a second set of experiments, semen was treated with 100 µM CGA, 1 mM H2O2 to induce OS, or H2O2+CGA; untreated samples were used as controls. Then, sperm motility, DNA integrity by the acridine orange test, F2-IsoPs and Nrf2 mRNA expression by RT-PCR were quantified. In CGA-treated specimens, rapid progressive sperm motility was increased (p < 0.01) and F2-IsoP levels decreased (p < 0.001) versus controls. The increase of F2-IsoP levels and DNA damage and the decrease of sperm motility after H2O2 treatment was reversed in the presence of CGA, which upregulated Nrf2 mRNA expression. These findings contributed to clarifying CGA’s antioxidant activity and highlighted the positive impact of CGA on sperm progressive motility, suggesting also a possible mechanism of action based on the Nrf2 pathway. CGA can be useful during human semen handling procedures in the laboratory and in optimizing the recovery of motile spermatozoa through selection techniques during assisted reproductive technology protocols.

Peroxisome Proliferator-Activated Receptor Gamma (PPARγ) is a regulating agent in antioxidant response also involved in controlling inflammation. The impact of varicocele and urogenital infections on sperm PPARγ expression was studied. The PPARγ gene expression was investigated in spermatozoa of 26 normozoospermic men grouped according to their clinical conditions: normal semen parameters (N), normal semen parameters and varicocele (N + V), and normal semen parameters and urogenital infections (N + UI). Sperm PPARγ expression was correlated with F2-isoprostanes (F2-IsoPs), as markers of lipid peroxidation, and Resolvin D1 (RvD1), a pro-resolving mediator in inflammation. Sperm PPARγ expression was evaluated through comparative real-time PCR, and F2-IsoPs and RvD1 were quantified in the seminal plasma via GC/NCI-MS/MS and immunoassay, respectively. PPARγ expression correlates positively with sperm morphology and vitality and negatively with F2-IsoPs and RvD1. Sperm morphology positively correlates with vitality and negatively with F2-IsoP and RvD1 levels. Despite the normozoospermia in the three examined groups, sperm morphology and PPARγ expression were significantly reduced in N + V and N + UI groups compared to the N group. Additionally, F2-IsoP and RvD1 levels were elevated in N + V and N + UI patients. These data suggest that PPARγ expression is compromised by inflammation and lipoperoxidation, providing new insights to further explore new possibilities of targeted treatment of male infertility.

This study aims to investigate the role of ferroptosis, an iron-dependent form of regulated cell death, in male infertility. The motivation behind this research stems from the increasing recognition of oxidative stress and iron metabolism dysregulation as critical factors in male reproductive health. In this study, 28 infertile patients (grouped by the presence of urogenital infections or varicocele) and 19 fertile men were selected. Spermiograms were performed by light microscopy (WHO, 2021). Testosterone, ferritin, transferrin-bound iron, transferrin, and F2-isoprostanes (F2-IsoPs) were detected in seminal plasma. Glutathione peroxidase 4 (GPX4) and acyl coenzyme A synthetase long chain family member 4 (ACSL4) were also assessed in sperm cells using enzyme-linked immunosorbent assays (ELISA). All the variables were correlated (statistically significant Spearman’s rank correlations) in the whole population, and then the comparison between variables of the different groups of men were carried out. Seminal ferritin and transferrin positively correlated with seminal F2-IsoPs, which had positive correlations with ACSL4 detected in sperm cells. Ferritin and ACSL4 negatively correlated with the seminal parameters. No correlation was detected for GPX4. Comparing the variables in the three examined groups, elevated levels of ACSL4 were observed in infertile patients with urogenital infections and varicocele; GPX4 levels were similar in the three groups. These results suggested a mechanism of ferroptosis, identified by increased ACSL4 levels and the occurrence of lipid peroxidation. Such events appear to be GPX4-independent in reproductive pathologies such as varicocele and urogenital infections.

Sperm cryopreservation is an important technique in preserving male fertility. Several methods for semen and sperm cryopreservation are available; however, the quality of thawed spermatozoa is poor and this is due to different mechanisms during the freezing-thawing process, including temperature changes, crystal ice formation, osmotic stress and oxidative stress (OS). OS is the result of an overproduction of reactive oxygen species (ROS) that, if present in high concentration, can damage the cellular structures and impair sperm function. Modulation of OS is an important issue in human sperm freezing. A large group of antioxidant molecules is used in cryopreservation processes as a pharmacological strategy to counteract the oxidizing effects of preservation procedures and thus protect sperm quality. The main body of the review comprises the analysis of different studies, starting from 2000 up to the present, dealing with the effects of different antioxidant compounds, including natural extract, used as supplement of cryopreservation media. Many studies have reported several beneficial effects of antioxidants that are added during freezing-thawing protocols on sperm cryo-damages; however, these improvements are not always evident. Over the past decade, the attention has been mainly focused on the phytoextracts and natural extracts. Phytoextracts can be obtained by waste products, which are a rich source of compounds with strong antioxidant activity. Because these by-products can be used in the industrial, cosmetic, nutraceutical, and human and animal reproductive fields, this topic of research is worthwhile implementing. The freezing and thawing protocols still have many pitfalls and the quality of thawed spermatozoa is not satisfactory. For this reason, new strategies to minimize cyodamages and to increase sperm cryostability are advisable to guarantee better sperm function and survival, permitting successful future clinical application.

This paper presents gaming as an alternative way of transmitting information compared to traditional academic methods. New technologies provide an opportunity, and in particular, virtual reality gaming allows users to have an immersive, personifying, intuitive, and engaging experience useful in visualizing complex concepts. Questionnaires were administered to secondary school students to extrapolate relevant information regarding the role of diet on sperm quality following the experience of the virtual reality game “Oxistress.” This game allows participants to understand in a playful and recreational way how the presence of reactive oxygen species can damage spermatozoa and how adequate diet choices provide ammunition in the form of antioxidants and omega-3 fatty acids to protect and repair spermatozoa. The results suggest that virtual reality (VR) may represent an important way to improve knowledge of scientific topics. Further studies will be necessary, involving larger populations with different basic knowledge as well as checks on whether the information acquired remains in memory after months.

The Special Issue “Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies”, published by Antioxidants and led by us (https://www [...]

Lipid mediators from fatty acid oxidation have been shown to be associated with the severity of Krabbe disease (KD), a disorder linked to mutations in the galactosylceramidase (GALC) gene. This study aims to investigate the effects of n-3 polyunsaturated fatty acid (PUFA) supplementation on KD traits and fatty acid metabolism using Twitcher (Tw) animals as a natural model for KD. Wild-type (Wt), heterozygous (Ht), and affected Tw animals were treated orally with 36 mg n-3 PUFAs/kg body weight/day from 10 to 35 days of life. The end product of PUFA peroxidation (8-isoprostane), the lipid mediator involved in the resolution of inflammatory exudates (resolvin D1), and the total amount of n-3 PUFAs were analyzed in the brains of mice. In Tw mice, supplementation with n-3 PUFAs delayed the manifestation of disease symptoms (p < 0.0001), and in the bran, decreased 8-isoprostane amounts (p < 0.0001), increased resolvin D1 levels (p < 0.005) and increased quantity of total n-3 PUFAs (p < 0.05). Furthermore, total brain n-3 PUFA levels were associated with disease severity (r = −0.562, p = 0.0001), resolvin D1 (r = 0.712, p < 0.0001), and 8-isoprostane brain levels (r = −0.690, p < 0.0001). For the first time in a natural model of KD, brain levels of n-3 PUFAs are shown to determine disease severity and to be involved in the peroxidation of brain PUFAs as well as in the production of pro-resolving lipid mediators. It is also shown that dietary supplementation with n-3 PUFAs leads to a slowing of the phenotypic presentation of the disease and restoration of lipid mediator production.

Background: Krabbe disease is due to a deficiency of lysosomal enzyme galactosylceramidase, which leads to destruction of the myelin sheath around nerves in the brain and spinal cord. In addition, Krabbe disease is associated with neuroinflammation in which harmful amounts of lipids are produced. To assess the importance of the regulation of lipid metabolism in the pathogenesis and intervention options in Krabbe disease, the aim of this study was to identify a set of specific biomarkers in a mouse model of the disease and to analyze the correlation between each biomarker. Methods: In this study, fatty acid mediators were investigated in twitcher mice, a natural model of Krabbe disease, and the genotype was determined. Mass spectrometry was used to quantify F-2-isoprostanes and immune techniques were used to investigate F-2-isoprostanes, resolvin D1 (RvD1), peroxisome proliferator-activated receptor gamma, apelin, and the apelin receptor in the brains of heterozygous and affected homozygous mice and in wild-type control mice. Results: The results of molecular analysis showed that there was a reduction in peroxisome proliferator-activated receptor gamma in the brains of both heterozygous and affected homozygous mice (p < 0.001). In addition, in the brains of mice with Krabbe disease RvD1 levels were decreased (p < 0.001), oxidation of arachidonic acid was increased (p < 0.001) and low levels of apelin (p < 0.001) were associated with an increase in apelin receptor (p < 0.05). RvD1 and apelin levels were associated with disease severity (r = -0.638, p < 0.001 and r = -0.725, p < 0.001, respectively). Conclusion: Our results indicate that mutation of the galactosylceramidase gene is associated with altered homeostasis of fatty acid oxidative metabolism. These homeostatic alterations reflect the disease phenotype. Our findings highlight a relevant aspect of fatty acid metabolism in the Krabbe disease brain and support the view that fatty acid metabolism is an active player in the pathogenesis of this still incurable disease.