Therapeutic potential of Oxalis corniculata in circumventing Alzheimer's disease through in vitro and in silico investigations.

Background: Bupivacaine (BUP) acts as a local anesthetic, which is extensively used for clinical patients but could generate neurotoxicity in neurons. Tetramethylpyrazine (TET) exhibits strong neuron protective effects against neurotoxicity. Hence, we investigate the effect of TET on BUP-induced neurotoxicity in SH-SY5Y cells.Methods: CCK-8 assay was used to detect cell proliferation in SH-SY5Y cells. In addition, Western blotting was used to examine Bax, Bcl-2, active caspase 3, LC3II, Beclin 1 and p-62 protein levels in cells. Moreover, ELISA assay was used to detect the levels of total glutathione (GS), superoxide dismutase (SOD) and malondialdehyde (MDA) in cells.Results: In this study, we found that TET attenuated the neurotoxicity of BUP on SH-SY5Y cells. Meanwhile, TET alleviated BUP-induced apoptosis in SH-SY5Y cell via decreasing the expressions of active caspase-3 and Bax and increasing the expression of Bcl-2. In addition, monodansylcadaverine staining assay and Western blotting results confirmed that TET induced autophagy in SH-SY5Y cells via increasing the LC3II/I and Beclin 1 levels. Furthermore, TET attenuated BUP-induced oxidative damage in SH-SY5Y cells via upregulation of the levels of total GS and SOD and downregulation of the level of MDA. Interesting, the protective effects of TET against BUP-induced neurotoxicity in SH-SY5Y cells were reversed by autophagy inhibitor 3-methyladenine (3MA).Conclusion: These data indicated that TET may play a neuroprotective role via inhibiting apoptosis and inducing autophagy in SH-SY5Y cells. Therefore, TET may be a potential agent for the treatment of human neurotoxicity induced by BUP.

Regular Coffee Intake is Related to Increased Sperm Motility and Antioxidant Levels in Infertile Men

Study question Do increased levels of oxidative stress (OS) and seminal antioxidant affect semen quality? Summary answer Increased OS measured by MiOXSYS negatively affected sperm concentration, motility, and total motile sperm count (TMSC). Reduced levels of seminal glutathione are associated with OS. What is known already The enzymatic seminal antioxidants superoxide dismutase (SOD) and catalase (CAT), and the non-enzymatic antioxidant glutathione possess important roles in the defense against oxidative damage and form the first line of defense against reactive oxygen species (ROS). It is known from the literature, that excessive levels of ROS are highly implicated in the pathogenesis of male infertility. The level of oxidative stress can be estimated by MiOXSYS Analyzer (Aytu BioScience Inc.) expressing the ‘oxidative reduction potential’ (ORP). Assessing ORP is one of the latest advancements in male infertility diagnostics, providing sensitive detection of increased OS in semen. Study design, size, duration This correlation study aims to explore the level of OS in a small cross-section of potentially infertile Danish males. Eighty-one (81) males from the Fertility Clinic at Odense University Hospital delivered a semen sample. The level of OS was compared to semen parameters and levels of SOD, CAT, and glutathione. Participants/materials, setting, methods Semen samples were collected and analyzed according to 2010 WHO criteria (concentration, volume, and %motility). TMSC was included additionally. A direct measure of OS was performed on raw sperm using MiOXSYS. Three assays were applied (Sigma-Aldrich and Abcam) to measure the enzymatic activity of SOD and CAT, and the level of total glutathione in the sperm samples. Spectrophotometric analysis of the absorbance in a plate reader was used to quantify the biochemical level of antioxidants. Main results and the role of chance Results showed that 43.2% of all study participants had increased levels of OS. Statistical analysis revealed a significant correlation (p< 0.01) between increased OS and reduced concentration, motility, and TMSC in the ejaculates. When adjusted for smoke, motility and TMSC were found to be significant (p < 0.05). Adjusting for weekly alcohol intake and body mass index (BMI) did not affect the correlation. Collectively, the results demonstrated that sperm abnormalities are related to increased levels of OS, suggesting OS to be a mediator of male fertility potential. Furthermore, a negative correlation (p< 0.05) was found between the reduced level of seminal glutathione and increased levels of OS. The finding suggests that OS is increased in the spermatozoa when the level of glutathione is low. No significant correlation between OS and reduced SOD or CAT was found. A potential inverse relationship between the effect of SOD on CAT and GSH levels was investigated, but no significant relationship was identified. There were no statistical indications that the level of SOD is associated with the levels of CAT and GSH. Limitations, reasons for caution Morphology was not included as a part of the WHO criteria due to time limitations. DNA fragmentation analysis was not included, but future research should be devoted to this aspect. Wider implications of the findings The findings support a suggestion of an association between high OS levels in semen and reduced semen quality. It was expected that at least 30% of men would show excessive OS in sperm. The study population of the Fertility-Clinic showed higher OS than what was presented in the general population. Trial registration number Not applicable

Autophagy is an evolutionary conserved catabolic process that ensures continuous removal of damaged cell organelles and long-lived protein aggregates to maintain cellular homeostasis. Although autophagy has been implicated in amyloid-β (Aβ) production and deposition, its role in pathogenesis of Alzheimer's disease remains elusive. Thus, the present study was undertaken to assess the cytoprotective and neuroprotective potential of autophagy on Aβ-induced oxidative stress, apoptosis and neurotoxicity in human neuroblastoma SH-SY5Y cells. The treatment of Aβ1-42 impaired the cell growth and redox balance, and induced apoptosis and neurotoxicity in SH-SY5Y cells. Next, the treatment of rapamycin (RAP) significantly elevated the expression of autophagy markers such as microtubule-associated protein-1 light chain-3 (LC3), sequestosome-1/p62, Beclin-1, and unc-51-like kinase-1 (ULK1) in SH-SY5Y cells. RAP-induced activation of autophagy notably alleviated the Aβ1-42-induced impairment of redox balance by decreasing the levels of pro-oxidants such as reactive oxygen species, lipid peroxidation and Ca2+ influx, and concurrently increasing the levels of antioxidant enzymes such as superoxide dismutase and catalase. The RAP-induced autophagy also ameliorated Aβ1-42-induced loss of mitochondrial membrane potential and apoptosis. Additionally, the activated autophagy provided significant neuroprotection against Aβ1-42-induced neurotoxicity by elevating the expression of neuronal markers such as synapsin-I, PSD95, NCAM, and CREB. However, 3-methyladenine treatment significantly exacerbated the neurotoxic effects of Aβ1-42. Taken together, our study demonstrated that the activation of autophagy provided possible neuroprotection against Aβ-induced cytotoxicity, oxidative stress, apoptosis, and neurotoxicity in SH-SY5Y neuronal cells.

In this study, a high-impact trauma (HIT) device was used for inducing moderate traumatic brain injury (TBI) in Drosophila melanogaster. Mechanical injuries in flies caused by rapid acceleration and assertion produce symptoms characteristics of TBI in humans.Docking studies were carried out to check the binding affinity of the drug toward the receptors. Various oxidative stress parameters, catalase level, glutathione level, superoxide dismutase (SOD) level, malondialdehyde (MDA), and nitric oxide levels, were measured. The mortality index and neuroprotective potential were carried out in TBI in D. melanogaster models.In the current study, there was an increase in oxidative stress following TBI as evidenced by a significant decrease in the catalase, glutathione, and SOD levels and increase in the level of MDA and nitric oxide after 24 hours. Antioxidant enzymes, catalase and glutathione peroxidase, have a dominant role in TBI. Docking studies were carried out on estrogen receptor 1 (pdb: 1TVO and 1UOM) and NDMA receptor (pdb: 3QEL) as agonist showing the binding affinity of the drug toward the receptors. In comparison to the vehicle-treated group, there was a dose-dependent significant increase in the SOD level and percentage climbing along with a decrease in the MDA level and total protein content. The mortality index was also observed at three concentrations of ranolazine (1, 2, and 4 mg/mL) in D. melanogaster homogenate. These findings suggest that ranolazine has a good neuroprotective potential in the treatment of TBI in the D. melanogaster model.Present study concluded the scientific evaluation of neuroprotective potential of ranolazine in the treatment of TBI in the D. melanogaster model.

We conducted a prospective study to comparatively evaluate serum levels of malondialdehyde, an oxidative stress indicator, and the antioxidant enzymes glutathione, catalase, and superoxide dismutase in patients with Bell palsy. Our study population was made up of 30 patients with Bell palsy—15 men and 15 women, aged 25 to 68 years (mean: 50.4)—who were seen in the Department of Otorhinolaryngology at a tertiary care medical center. For comparison purposes, we recruited 26 healthy age- and sex-matched controls—16 men and 10 women, aged 40 to 67 years (mean: 54.3). Serum samples were obtained from all participants before the initiation of steroid treatment to the Bell palsy patients. A correlation was sought between demographic data and serum levels of malondialdehyde, glutathione, catalase, and superoxide dismutase. We found that the serum level of malondialdehyde was significantly higher in the Bell palsy group and that the levels of glutathione, catalase, and superoxide dismutase were all significantly lower (p < 0.001 for all comparisons). Correlation analysis revealed that only superoxide dismutase levels were positively correlated with age (r = 0.347, p = 0.009). We suggest that oxidative stress and antioxidant mechanisms may be involved in the pathogenesis of Bell palsy. In this context, serum levels of malondialdehyde, glutathione, catalase, and superoxide dismutase may serve as biomarkers in the diagnosis and follow-up of Bell palsy. Confirmation of the validity, reliability, and reproducibility of these findings necessitates further prospective, randomized clinical trials in larger populations.

Deregulated lncRNA MAGI2-AS3 in Alzheimer's disease attenuates amyloid-β induced neurotoxicity and neuroinflammation by sponging miR-374b-5p.

To explore the protective effect of Cornus officinalis extracts on Alzheimer’s disease cell injury models and its underlying molecular mechanism. Pheochromocytoma cells were treated with 20 μM amyloid beta-peptide 25-35 to establish an Alzheimer’s disease cell injury model in vitro, which was recorded as amyloid beta-peptide 25-35 group. Cells affected by amyloid beta-peptide 25-35 were treated with different-doses of Cornus officinalis extracts and recorded as low-dose Cornus officinalis extracts group, medium-dose Cornus officinalis extracts group and high-dose Cornus officinalis extracts group. Pheochromocytoma cells transfected with si-NC/si-long non-coding RNA Rpph1, treated with 20 μM amyloid beta-peptide 25-35 and 80 mg/ml of Cornus officinalis extracts were recorded as high-dose Cornus officinalis extracts+si-NC group, high-dose Cornus officinalis extracts+si-long non-coding RNA Rpph1 group. Cell viability and apoptosis were examined using cell counting kit-8 assay and flow cytometry. Protein expression was tested by Western blot. Malondialdehyde, superoxide dismutase and catalase levels were assessed by measuring cell oxidative stress. Amyloid beta-peptide, tumor necrosis factor-alpha, interleukin-6 and interferon gamma levels were examined using enzyme-linked immunosorbent assay. Long non-coding RNA Rpph1 expression was detected using quantitative reverse transcriptase polymerase chain reaction. Amyloid beta-peptide 25-35 treatment decreased Pheochromocytoma cell viability, CyclinD1, superoxide dismutase, catalase and long non-coding RNA Rpph1 levels, while increased apoptosis rate, cleaved-caspase-3, malondialdehyde, amyloid beta-peptide, tumor necrosis factor-alpha, interleukin-6 and interferon gamma levels. After treatment with different-doses of Cornus officinalis extracts, cell viability, CyclinD1, superoxide dismutase, catalase and long non-coding RNA Rpph1 levels were enhanced, while apoptosis rate, cleaved-caspase-3, malondialdehyde, Aβ, tumor necrosis factor-alpha, interleukin-6 and interferon gamma levels were reduced in pheochromocytoma cells treated with amyloid beta-peptide 25-35. Downregulation of long non-coding RNA Rpph1 reversed the inhibitory effect of Cornus officinalis extracts on cell injury. Cornus officinalis extracts relieved amyloid beta-peptide 25-35-induced nerve cell injury by upregulating long non-coding RNA Rpph1, suggesting that Cornus officinalis extracts might be used in Alzheimer's disease treatment.

DHA, EPA and their combination at various ratios differently modulated Aβ25-35-induced neurotoxicity in SH-SY5Y cells

Introduction: Recent studies have demonstrated that alpha-lipoic acid (ALA) has a strong antioxidant property and it exerts neurotrophic effects on the peripheral nerves. In this study, we investigated potential effects of ALA on secondary injury mechanisms as well as on apoptosis.Methods: Forty Sprague-Dawley rats were equally divided into 5 groups, as follows: laminectomy (control), laminectomy + trauma (Trauma), laminectomy + posttraumatic methylprednisolone [MP] (Trauma + MP), laminectomy + posttraumatic ALA (Trauma + ALA), laminectomy + posttraumatic MP and ALA (Trauma + MP + ALA). Yasargil aneurysm clip method was used to induce the spinal cord injury. Twenty-four hours after the procedure the rats were sacrificed. Spinal cord samples were harvested to analyze malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) levels, as well as histopathological characteristics.Results: The Kruskal-Wallis test (95% confidence level, p < 0.05) showed a statistically significant difference between the groups in MDA (p = 0.006), CAT (p = 0.000), and SOD (p = 0.001) levels. Pairwise comparisons, with Bonferroni correction, of control and Trauma group with the other groups, revealed a significant difference in CAT and SOD levels. Overall, our results showed that ALA administration significantly decreased MDA levels in Trauma + ALA compared to the same effect of MP in Trauma + MP group. Furthermore, ALA administration increased SOD and CAT levels in Trauma + ALA group. The combined use of ALA and MP demonstrated synergistic effects and yielded even more significant results.Conclusions: A combined use of ALA and MP might provide a benefit in patients exposed to spinal cord injury. However, at present, further studies are required to confirm these results.

This study aimed to investigate the neuroprotective role of probiotics and 1,25-dyhydroxyvitamin D3 (calcitriol) against neurotoxicity on rotenone-induced human neuroblastoma cell line SH-SY5Y. Rotenone was administered to induce neurotoxic effects in SH-SY5Y cells. Calcitriol and probiotics were administered at different concentrations as pre- and post-treatment. The thiazolyl blue tetrazolium bromide (MTT) assay was performed to measure cell viability. Intracellular protein levels of antioxidant enzymes (protein tyrosine kinase (PTK), superoxide dismutase (SOD), glutathione peroxidase (GSH), glutathione reductase (GSR), and catalase (CAT)) were determined by the enzyme-linked immunosorbent assay (ELISA). Rotenone (150 nM) reduced (p < 0.001) cell viability compared to control cells. Single and combined pretreatments with probiotics (0.01 mg/ml, 0.05 mg/ml, and 0.1 mg/ml) and calcitriol (1.25 µM, 2.5 µM, and 5 µM) increased (p < 0.05) cell viability compared to rotenone group. In the pre- and post-treatment design, all treatment groups increased the SOD and GSH levels and decreased the GSR levels compared to rotenone. None of the pretreatments reversed the PTK levels (except probiotics: 0.01 mg/ml). Calcitriol (2.5 µM) increased the CAT levels in pretreatment design, and probiotics (0.05 mg/ml and 0.1 mg/ml) increased CAT levels in post-treatment design compared to rotenone group. Calcitriol and probiotics protect against rotenone-induced neurotoxicity in SH-SY5Y cells by decreasing reactive oxygen species (ROS) and increasing antioxidant enzyme parameters. These neuroprotective effects of calcitriol and probiotics against rotenone-induced dopaminergic neurotoxicity provide an experimental basis for their potential clinical use in the treatment of Parkinson’s disease (PD).

Methylparaben protects 6-hydroxydopamine-induced neurotoxicity in SH-SY5Y cells and improved behavioral impairments in mouse model of Parkinson's disease

P-844: Relationship between seminal viscosity and oxidative stress in infertile men

Humans are most exposed to the heavy metal cadmium (Cd), known as neurotoxic. However, it is unclear how selenium (Se) protects neurons from damage caused by increased Cd-induced neurotoxicity in SH-SY5Y cells and how the TRPM2 channel functions in this process. In this study, we examined the impact of Se on CdCI2-induced oxidative neurotoxicity and cell death in SH-SY5Y cells by modifying the TRPM2 channel. The Se and TRPM2 channel antagonist 2-APB was added to prevent CdCI2-induced neurotoxicity in SH-SY5Y cells. Cell viability rate was determined between groups by CCK-8 assay. GSH, MDA, and ROS levels were determined in the cells with ELISA kits. Our results showed that the TRPM2 channel plays a vital role in forming CdCI2-induced damage to cells by using the TRPM2 antagonist in the study. We also observed that Se reduced CdCI2-induced neurotoxicity by reducing TRPM2 channel activation by suppressing oxidative stress of cells. We conclude that Se therapy and TRPM2 channel blocking can reduce CdCI2-induced neurotoxicity based on our investigation, which examined the protective impact of Se and the involvement of the TRPM2 channel in CdCI2-induced SH-SY5Y cells for the first time. FULL TEXT PDF

Rotenone-induced neurotoxicity in SH-SY5Y cells is an essential hallmark of neurodegenerative diseases like Alzheimer's disease (AD) and Parkinson's disease (PD). β-Caryophyllene (BCP), a cannabinoid receptor 2 (CB2) agonist, has anti-inflammatory, antioxidant, and cytoprotective efficacy. The involvement of the GSK-3β/NRF2/HO-1 axis in neuroprotection has garnered attention as a possible mechanism for BCP to exhibit multitargeted neuroprotective effects. Hence, this study investigates the effects of BCP against rotenone-induced neurotoxicity and apoptosis in SH-SY5Y cells, focusing on the involvement of the GSK-3β/NRF2/HO-1 signaling pathway. Initially, we performed the in silico molecular docking of BCP with GSK-3β, NRF2, and HO-1 proteins to ensure the degree of binding affinities. The in vitro MTT assay was performed to evaluate cell viability, followed by the assessment of biomarkers such as LDH leakage, oxidative stress, reactive species, caspase 3 activity, pro-inflammatory markers, and GSK-3β, NRF2, and HO-1 proteins in BCP, as well as specific receptor modulators (chir98023 and quercetin) against the rotenone pre-treated cells. In silico molecular docking studies revealed that BCP exhibits a strong binding affinity for GSK-3β, NRF2, and HO-1 proteins. Also, in vitro studies revealed that BCP (100µg/ml), as compared to the rotenone-treated group, significantly restored cell viability (72%). Moreover, BCP significantly modulates cell cytotoxicity (LDH leakage), pro-apoptotic, pro-inflammatory, reactive species, and oxidative stress markers. Molecular docking established robust binding affinities of BCP with GSK-3β, NRF2, and HO-1 proteins. Furthermore, protein estimation by ELISA confirmed the BCP-mediated modulation of these pathways. These findings suggest that BCP protects SH-SY5Y cells from rotenone-induced neurotoxicity, offering a potential therapeutic candidate for neurodegenerative diseases like AD.