LDH Enzyme Research Articles

Nandrolone decanoate induces heart injury via oxidative damage and mitochondrial apoptotic pathway by regulation of TLR4/NF-κB/NLRP3 axis in male rats: The rescue effect of N-acetylcysteine.

Myocardial apoptosis is a leading cause of damage in cardiac tissues of nandrolone (ND) treatment. However, its molecular mechanism is not fully understood. This study aims to investigate the effect of ND with or without N -acetylcysteine (NAC) treatment on oxidative damage and TLR4/NF-κB /NLRP3 signaling pathway in the heart of male rats. Eighteen male Wistar rats with a weight range of 220 ± 10 g were selected. They were divided into three groups (n = 6): control (C) group, ND group, NAC + ND group. After six weeks of treatment, the TUNEL staining indicated that ND increased the number of apoptotic cells in the hearts of male rats. The molecular analysis demonstrated that ND exposure resulted in increased protein levels of cytochrome c, c-Caspase-3/p-Caspase-3 ratio, p53, TLR4, NF-κB, NLRP3, and 8-OHdG with a concomitant up-regulation of LDH and CK-MB enzymes activity in the heart tissue compared to the C group. Our findings suggested that ND can cause damage to heart tissue via induction of DNA damage, apoptosis, and probably TLR4/NF-κB/NLRP3 signaling pathway plays a crucial role in this process. It also demonstrates that these negative effects of ND can be reduced by using NAC treatment as an antioxidant and anti-inflammatory agent.

Antioxidant Enzyme Responses in Cladoceran Moina macrocopa under Oxidative Stress

Oxygen is the cornerstone of the life of all living organisms, as it plays a pivotal role in regulating the bio-physiological functions of the organism that are necessary to sustain its existence. Oxidative stress/ hypoxia is a condition of inadequate oxygen that disrupts the harmony of an organism's life by impairing its growth, puts the organism's survival at risk, and destabilizes the ecosystem. Like all other organisms Moina, a small cladoceran is susceptible to the alteration in dissolved oxygen levels. Here we report that, in response to hypoxia, Moina adjusts their antioxidant enzyme activities as a mechanism of adaptation. Significant changes were observed in the levels of LDH, GST, and SOD antioxidant enzymes of Moina under hypoxia (p=0.001053, GST p=0.0010053, SOD p=0.0015). These results can contribute to wider research on environmental stress tolerance by aquatic organisms and will assist in the conservation of species like Moina. Moina, being an ecological indicator, this research will help in environmental monitoring using it as a model organism.

Murine model evaluation of biochemical and tissue-damage induced by Bothrops asper (Nauyaca) snake venom

Introduction: Venomous snakes Bites are a serious public health issue, and in Mexico, Bothrops asper is responsible for the majority of viper bites. Objective: To evaluate, in a mouse model, the effect of Bothrops asper venom snake on liver enzymes LDH, AST, and ALT, as well as to assess its effect on renal and hepatic histology. Methods: The LD50 was determined using a dose-response curve after intramuscular administration of increasing doses of the venom to mice (0.5, 1, 2.5, 5, and 10 mg of protein/kg). Subsequently, the mice were administered the LD25 value of the venom (or saline solution for control), and 24 hours after the injection, LDH, AST, and ALT enzymes were measured in the blood, and the liver and kidneys were collected from each animal. Results: Results: Animals treated with poison (LD25 of 1 mg of protein/kg) had greater activity of AST, ALT and LDH enzymes, structural damage in the cortex and renal medulla, as well as in the liver after 24 hours of administration in the experimental group Conclusion: B. asper venom causes an increase in LDH and transaminase enzymes, as well as renal and hepatic histopathological alterations.

Withdrawal Effects on Antioxidative and Histochemical Deficits in Acetaminophen Induced Neurotoxicity in Cerebellar Cortex of Adult Wistar Rats

Introduction: Neurotoxicity refers to the harmful effects of chemical, biological, or physical agents on the nervous system. Acetaminophen, a common pain reliever whose abuse has been linked to neurotoxicity at high doses, causing oxidative stress and neuronal damage. Aim: The present study aimed to assess the withdrawal effects of Acetaminophen use on the Cerebellar cortex of adult wistar rats. Methodology: Thirty (30) Adult wistar rats weighing 200±50g were assigned into three groups (n=10) of Control (C), Treatment 1(T1) and Treatment 2 (T2). The control group C received distilled water, while Treatment groups T1 and T2 received 200mg/kg of acetaminophen for 4 weeks. However, treatment group T2 were allowed a 2-week acetaminophen withdrawal period. At the end of exposure, all animals were sacrificed via cervical dislocation and eventual removal of cerebellar specimens which were processed for some Neurohistochemical staining reactions as well as Biochemical Quantifications of LDH enzyme and Antioxidative stress markers (SOD & CAT). Quantitative analyses of all data obtained including Body weight, brain weight and Cerebellum weight were analysed using GraphPad® (version 8) and plotted using ANOVA followed by Tukey's multiple comparisons test. Significance was set at p<0.05* (95% confidence interval). Result: Results showed a significant increase in body weight (P=.05) and a decrease in cerebellum weight (P=.01) in both acetaminophen groups. SOD and CAT activity decreased significantly (P=.01) in the T1 groups while T2 groups shows a significant increase, LDH activities decreased significantly (P=.01) in both groups. The Neurohistochemical findings showed severe degeneration, loss of the Purkinje neurons and poorly differentiated DNA on the T1 group compared to control group C while the withdrawal group showed onset of regenerative changes in neurons improved differentiation in DNA stained. Conclusion: The study's findings suggest that long-term acetaminophen use can lead to neurotoxic effects on the cerebellum, but partial recovery is possible upon withdrawal.

Comparative assessment of the most important markers of equine myopathy in connection with creatine kinase activity

Purpose: to assess the activity of AST, LDH and HBDH enzymes in relation to different levels of total creatine phosphokinase activity to understand the degree of their diagnostic value in determining the severity of myopathy in horses.Materials and methods. For the study, statistical processing of a data array of the results of a biochemical study of blood serum from 247 horses was carried out. Taking into account the reference intervals of CPK activity, all results were ranked by increasing creatine phosphokinase activity and divided into four groups. In each group, the average values for AST, LDH and HBDH were calculated, and the degree of significance of intergroup differences was calculated using the Student's t test. The correlation between the activity of the enzymes AST, LDH and HBDH with the activity of CPK in each group was also assessed.Results. An increase in the activity of creatine phosphokinase in horses by 59,4—409,6 % is accompanied by an increase in the activity of the enzymes AST by 14,5—77,2 %, LDH by 17,8—33,0 % and HBDH by 17,1—27,1 %. When comparing Pearson's correlation coefficients across the entire study sample, a significant direct relationship is determined between CPK and AST and a moderate direct relationship when comparing CPK with LDH and HBDH. Calculation of the Pearson coefficient in each of the studied groups showed different degrees of correlation between the creatine phosphokinase indicator and other enzymes, but in all cases positive. A study of the percentage contribution of HBDH to the total lactate dehydrogenase activity showed an inversely proportional relationship with respect to the activity of creatine phosphokinase, which indicates massive muscle damage with switching energy metabolism in them.Conclusion. The conducted studies made it possible to more deeply understand the cause-and-effect relationships of changes in the activity of enzymes that are specific to muscle tissue in connection with the severity of myopathy. The data obtained can assist veterinary specialists in analyzing the results of biochemical blood tests in horses.

Effect of crude ethanolic seed extract from Mucuna pruriens on proliferation, apoptosis, and cell cycle arrest in gastric adenocarcinoma (AGS) cells

Abstract Background Gastric cancer is a prevalent form of malignancy among many common carcinoma cases globally. This study was designed to assess the anticancer potential of the crude ethanolic seed extract from Mucuna pruriens against the gastric cancer cell line (AGS). Various assays were employed to assess the anticancer properties, including examinations of cell viability, nuclear morphology, apoptosis using AO/EB staining, changes in mitochondrial membrane potential, lactate dehydrogenase activity, DNA fragmentation, and cell cycle arrest. Results The crude extract exhibited significant anticancer activity against the human gastric cancer cell line (AGS), as determined by the MTT assay, with an inhibition concentration (IC50) of 600 µg/mL at 24 h. Distinct cellular and nuclear morphological changes were observed with different concentrations of crude ethanolic seed extract. The LDH release assay reveals cell death in AGS cells, as evidenced by a significant increase in the release of LDH enzyme. DNA fragmentation analysis and flow cytometry results indicate that the extract induces chromatin condensation, apoptotic cell death, and cell cycle arrest at the G0/G1 phase in the AGS cancer cell line. These results highlight the potential therapeutic advantages of Mucuna pruriens seed extract against gastric cancer cells. Conclusion This study could pave the way for identifying diverse natural bioactive compounds sourced from Mucuna pruriens seed, leading to the development of novel drug with potential anticancer properties.

Short-Term Atorvastatin Therapy in Healthy Individuals Results in Unaltered Plasma MMP Levels and Disrupted MMP-7 Correlation with Blood Lipids and Blood Count-Derived Inflammatory Markers.

Background: Matrix metalloproteinases (MMPs) play an important role in the pathophysiology of atherosclerosis. Reportedly, statins can decrease MMP activity in patients with atherosclerotic cardiovascular disease, but this effect has not been studied in healthy individuals. Methods: MMPs 2, 7, and 9 and several other parameters were measured before and after a four-week course of moderate-dose atorvastatin (20 mg/day) in 21 healthy individuals. Results: Atorvastatin treatment resulted in lower total cholesterol, LDL-cholesterol, non-HDL-cholesterol, and triglycerides (p < 0.001 for all), but higher levels of plasma enzymes AST, ALT, CK, and LDH (p < 0.05 for all). No effect of atorvastatin on plasma MMP median concentrations was recorded. Before treatment, moderate positive significant correlations were found between MMP-7 and age, blood lipids, and blood count-derived inflammatory markers. Pre-treatment MMP-7 was best predicted by the total cholesterol-to-HDL cholesterol ratio in a remnant cholesterol-weighted least squares regression model. After atorvastatin treatment, MMP-7 no longer correlated with these markers. Conclusions: While the effect of statins on plasma MMPs in atherosclerosis is controversial, short-term moderate-dose atorvastatin treatment does not seem to affect levels of MMPs 2, 7, and 9 in healthy individuals. However, an intriguing correlation between MMP-7 and atherosclerosis-related blood lipids and neutrophil-associated inflammatory biomarkers seems to be disrupted by atorvastatin independently of hsCRP, possibly via pleiotropic effects.

OsLdh7, a rice lactate dehydrogenase, confers stress resilience in rice under cadmium stress through NAD+/NADH regulation

Lactate dehydrogenase (Ldh, EC 1.1.1.27), an oxidoreductase enzyme catalyses the interconversion of pyruvate to L-lactate and vice-versa with concomitant oxidation and reduction of NADH and NAD+. The enzyme functions as a ROS sensor and mitigates stress response by maintaining NAD+/NADH homeostasis. In this study, we delineated the role of the Ldh enzyme in imparting cadmium stress tolerance in rice. Previously, we identified a putatively active Ldh in rice (OsLdh7) through insilico modelling. Biochemical characterization of the OsLdh7 enzyme revealed it to be optimally active at pH 6.6 in the forward direction and pH 9 in the reverse direction. Overexpression of OsLdh7 in rice cv. IR64, increased tolerance of the transgenic lines to cadmium stress compared to the wild type (WT) at both seedling and reproductive stages. The transgenic lines showed increased enzyme activity in the reverse direction under cadmium stress, attributed to elevated cytosolic pH resulting from increased calcium concentration. This increased NADH content is highly essential for functioning of the ROS scavenging enzymes, RbohD and MPK6. qPCR analysis revealed that the overexpression lines had increased transcript abundance of these genes indicating an effective ROS scavenging mechanism. Additionally, the overexpression lines showed an efficient cadmium sequestration mechanism compared to the WT by increasing the transcript levels of the vacuolar transporters of cadmium as well as total phytochelatin content. Thus, our findings indicated OsLdh7 imparts cadmium stress tolerance in rice through a two-pronged approach by mitigating ROS and sequestering cadmium ions, highlighting its potential for crop improvement programs.

Exosomes of mesenchymal stem cells and PRP restore spermatogenesis in the rat model of non-obstructive azoospermia.

Since available therapeutic approaches for chemotherapy-induced non-obstructive azoospermia (NOA) patients are not enough efficient, an urgent need for treatment alternatives is felt. This study shows that adipose tissue-derived mesenchymal stem cells-derived exosome (AD-Exo) treatment is more effective in ameliorating busulfan-induced NOA rat models compared to platelet-rich plasma (PRP). Patients with non-obstructive azoospermia (NOA) are unable to have their children. Therefore, there is an urgent need for additional treatment alternatives for these patients. Recently, novel treatments based on the exosomes derived from mesenchymal stem cells (MSCs) as the agents responsible for exerting the paracrine effects and consequently biological functions of MSCs are proposed. Besides, platelet-rich plasma (PRP) as a significant blood byproduct has been therapeutically applied in several male infertility studies. In this study, we compared the effects of PRP and exosome treatment on spermatogenesis restoration in NOA rat models. Exosomes and PRP were isolated from the adipose tissue-derived MSCs (AD-MSCs) collected from conditioned medium and peripheral blood of human volunteers, respectively. Non-obstructive azoospermia (NOA) induction was done through two doses of busulfan at a 21-day interval. Thirty-five days after NOA induction, intratesticular injection of AD-MSCs-derived exosome (AD-Exo), PRP, and PBS was performed. The control group did not receive any treatment. Two months later, the rats were euthanized for further analysis. Our results revealed that both AD-Exo and PRP treatments improved the size and weight of testis, modulated the expression level of Dazl, Ddx4, Stra8, Pwil1, and Ccna1, and ameliorated the serum level of LDH, SOD, and GR enzymes in NOA rats. Moreover, the AD-Exo group showed improved testosterone, GPx, MAD, and CAT serum levels, sperm motility, and protein levels of DAZL and DDX4. This investigation verified the more efficient effects of AD-Exo treatment in comparison to PRP in ameliorating busulfan-induced NOA rat models.

Melatonin regulates mitochondrial function to alleviate ferroptosis through the MT2/Akt signaling pathway in swine testicular cells

Increasing evidence has shown that many environmental and toxic factors can cause testicular damage, leading to testicular ferroptosis and subsequent male reproductive disorders. Melatonin is a major hormone and plays an vital role in regulating male reproduction. However, there is a lack of research on whether Mel can alleviate testicular cell ferroptosis and its specific mechanism. In this study, the results indicated that Mel could enhance the viability of swine testis cells undergoing ferroptosis, reduce LDH enzyme release, increase mitochondrial membrane potential, and affect the expression of ferroptosis biomarkers. Furthermore, we found that melatonin depended on melatonin receptor 1B to exert these functions. Detection of MMP and ferroptosis biomarker protein expression confirmed that MT2 acted through the downstream Akt signaling pathway. Moreover, inhibition of the Akt signaling pathway can eliminate the protective effect of melatonin on ferroptosis, inhibit AMPK phosphorylation, reduce the expression of mitochondrial gated channel (VDAC2/3), and affect mitochondrial DNA transcription and ATP content. These results suggest that melatonin exerts a beneficial effect on mitochondrial function to mitigate ferroptosis through the MT2/Akt signaling pathway in ST cells.

Myct1 Alleviates Hypoxia-Induced Dysfunction by Regulating Pericyte Reprogramming

As one of the main causes of death from cardiovascular diseases, myocardial infarction has brought a heavy burden to society. However, its underlying mechanism has not been elucidated. Irreversible contraction of pericytes will cause capillary contraction, resulting in microcirculatory disorder, which finally lead to no-reflow after myocardial infarction. In the current study, we used hypoxia to simulate the environment of myocardial infarction in vitro, and found that under hypoxia conditions, the contractility of pericytes was significantly enhanced, the apoptosis rate and the content of angiogenic factors was increased. Besides, a target gene of c-Myc, Myct1, could regulate pericytes reprogramming into endothelial cells. After reprogramming of pericytes, the contractile ability was reduced, and the ability to promote angiogenesis was also inhibited. Moreover, pericyte reprogramming significantly reduced the expressions of myocardial enzymes CK-MB and LDH, troponin TnT and inflammatory cytokine IL-6. In conclusion, the reprogramming of pericytes regulated by Myct1 could alleviate the dysfunction of pericytes, thereby inhibiting the expression of myocardial infarction markers, which was conducive to improving the phenomenon of no-reflow after myocardial infarction.

Repeated dose, 28-day oral toxicity study of curcumin, anthocyanins, and sodium nitrite in Wistar rats

ObjectiveThe goal of the present study was to examine the repeated dose 28-day oral toxicity of curcumin, anthocyanins, and sodium nitrite in Wistar rats. MethodsFor this purpose, forty-eight male Wistar rats were randomly divided into 8 groups (n = 6 each), encompassing untreated controls and experimental groups treated with curcumin, anthocyanins, and sodium nitrite. Three rats from each group were sacrificed by cervical dislocation under di-ethyl ether anesthesia after 2 and 4 weeks of therapy, respectively. Blood samples were collected for serum chemistry. All of the animals' livers, hearts, and kidneys were removed and sent for histopathological examination. ResultsAfter two weeks of inquiry, certain groups displayed higher hematological values, while others had lower values compared to the control group. AST, CK, and LDH enzyme activity were higher in groups 2–8, but urea concentrations were higher in groups 6 and 8. After four weeks, the Hb, MCH, and MCHC values in group 4 were greater, as were the WBC levels in groups 4 and 6, whereas other groups had lower MCV and WBC values. The weekly body weight gain was insignificantly different between treatment groups. Throughout the experiment, none of the animals perished. Male rats' liver, kidney, and heart underwent histopathological changes after ingesting curcumin, sodium nitrite, and anthocyanin. ConclusionBased on the findings, rats were more detrimental when curcumin, sodium nitrite, and anthocyanin were ingested together than when they were consumed individually, as evidenced by histopathological abnormalities in the liver, kidneys, and heart.

Study of The Effect of Boswellia Leaf Extract on The Activity of Cardiac Enzymes (ALP, AST, ALT, LDH) Using Spectroscopic Methods

This study investigates the impact of Boswellia carterii leaf extracts on serum enzyme activity in myocardial infarction (MI) patients. Employing a comparative analysis between 60 healthy individuals and 40 cardiac patients, we measured the serum activity of ALP, AST, ALT, and LDH enzymes. Our findings reveal elevated enzyme activity in MI patients. We explored the inhibitory effects of aqueous, alcoholic, and ethyl acetate extracts of Boswellia carterii on these enzymes, demonstrating non-competitive inhibition. The inhibition ratio of ALT, ALP, and AST post-treatment was quantitatively assessed. Statistical analysis was conducted using SPSS-26 with independent t-tests. This research highlights the potential therapeutic role of Boswellia carterii in modulating enzyme activity associated with cardiac events, suggesting avenues for novel treatments in cardiac care.

Evaluation of some proinflammatory cytokines and biochemical parameters in pre and postmenopausal breast cancer women

The study was planned to evaluate the differences in certain proinflammatory cytokines(IL-6, TNF-α) with CRP and biochemical parameters (E2, D3, LDH, GGT, TSB, Ca, Ph, uric acid), between women with pre- and postmenopausal breast cancer and seemingly healthy women in Iraqi women as controls; at medical city in teaching Oncology hospital,70 breast cancer patients women their ages ranged (47.51 ± 1.18) and 20 healthy women with age (44.45 ± 2.66) begun from September (2020) to February (2021). The aims of this study to investigate the evaluation of chemotherapy effects especially doxorubicin and cyclophosphamide only use in this study in pre and postmenopausal breast cancer women on proinflammatory cytokines(IL-6, TNF-α) with CRP and on biochemical parameters(E2, D3, LDH, GGT, TSB, Ca, Ph, uric acid) in pre and postmenapausal breast cancer women. The patients were divided into five groups and each group contains 14 patients women with breast cancer during pre and postmenopausal periods. The control groups were divided into 10 pre and 10 postmenopausal women(Fig. 1). The results of proinflammatory cytokines of and biochemical parameters in premenopausal groups were as the levels of IL-6 (pg/ml),TNF-α(pg/ml) and CRP (ng/ml) showed significant increase differences (P < 0.01)among breast cancer treated (BCT) groups in comparison with control groups,While the Liver enzymes GGT,LDH and TSB showed highly significant increase (P < 0.01) in BCT groups, Estrogen levels (pg/ml) and D3(ng/ml) increased significantly (P < 0.01)among BCT groups. Blood serum calcium and phosphorus with uric acid levels (mg/dl) showed significant difference (P < 0.01); While the result in postmenopausal of IL-6(pg/ml), TNF-α (pg/ml) and CRP (ng/ml) showed highly significant differences (P < 0.01)among BCT groups.While GGT(IU/L), LDH(IU/L) and TSB (mg/dl) enzymes were increased significantly (p < 0.01), Estrogen (pg/ml) and D3(ng/ml) levels showed significant increase (P < 0.01) among BCT groups.Blood calcium and phosphorus showed significant increase (P < 0.01) while uric acid was non-significant increase (P > 0.05).

In vitro characteristics of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and TLR7/8 ligand R848

The quality of the separated fractions in sex-sorted semen is very important for the success of the artificial insemination. This study aimed to evaluate some in vitro characteristics (DNA quantity, kinematic parameters and enzymes activity) of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and toll-like receptors (TLR)7/8 ligand R848. The ejaculates from six rams were collected by artificial vagina and subjected to a computer-assisted semen analysis (CASA). Total motility and percentage of the sperms with rapid and medium progressivity or non-progressivity in whole ejaculates and in X and Y fractions were analyzed. Activity of the enzymes ALP, GGT, CK, LDH and accumulation of lactate in the seminal plasma of ejaculates and in the environmental fluid of sexed spermatozoa were measured by biochemical analyzer. DNA was isolated from precipitated spermatozoa, and its quantity was measured. For both protocols the DNA mass from X-bearing fractions was higher, than from Y-bearing fractions. The high total motility of X- and Y-bearing spermatozoa as well as greater percent sperms with progressive motility were observed after use of BSA protocol. The application of TLR7/8 ligand R848 protocol led to reducing of Y-sperm motility and enhancement of non-progressivity in both fractions, which corresponded to the determined high amount of the extracellular lactate. For both methods, the significantly reduced activity of enzymes in the X and Y spermatozoa environmental fluids was established. Both protocols produce X- and Y-sperm fractions with satisfactory quality (over 80% total motility and over 50% rapid and medium progressive spermatozoa in each fraction).

Individual and combined impact of microplastics and lead acetate on the freshwater shrimp (Caridina fossarum): Biochemical effects and physiological responses

Microplastics and heavy metals pollution is recognised as a major problem affecting aquatic ecosystems. For this reason, this study aims to assess the toxicity of different concentrations of polyethylene microplastics (PE-MPs) (0.0, 500, and 1000 μg L−1) with a mean size of 15–25 μm and lead acetate Pb(C2H3O2)2 (0.0, 2.5, and 5 mg L−1), both individually and in combination, through the exposure of the freshwater grass shrimp, Caridinia fossarum for 15 days, focusing on microplastic interaction with co-occurring contaminants. After being exposed to both contaminants, either individually or in combination, significant alterations in numerous biochemical markers were observed. Specifically, exposure to lead acetate alone resulted in significant changes across ALP, AST, ALT, LDH, GGT, and BChE enzyme activity levels indicating hepatotoxicity and neurotoxicity. Also, Pb exposure led to alterations in total antioxidant capacity, MDA, total lipids, and glycogen contents, signalling the onset of oxidative stress. Exposure to PE-MPs alone led to changes in ALP, LDH, GGT, and BChE enzyme levels, and in MDA, total lipids, and glycogen samples' contents. Remarkably, the study observed increased bioaccumulation of lead acetate in samples treated with the combination, emphasizing the synergistic impact of PE-MPs on the toxicity of lead acetate. This synergy was also evident in AST and ALT enzyme activity levels and MDA contents. This underscores the necessity for measures to address both microplastic pollution and heavy metal contamination, taking into account the synergistic behaviour of MPs in the presence of concurrent contaminants.

The function of NLRP3 in anti-infection immunity and inflammasome assembly of common carp (Cyprinus carpio L.)

NLRP3 inflammasome can be activated by a variety of stimuli and plays an important role in protecting host from pathogen invasion and maintaining homeostasis. However, the activation mechanism of NLRP3 inflammasome in fish is still unclear. In the present study, the NLRP3 gene (CcNLRP3) was identified from common carp, which was 3069 bp in length and encoded a protein with five domains. Sequence analysis showed that NLRP3 was evolutionarily conserved, and CcNLRP3 was closely related to that in grass carp and zebrafish. Real-time PCR showed that CcNLRP3 was widely expressed in various immune-related tissues of healthy common carp, and significantly increased after stimulation with E. tarda, A. hydrophila and Cyprinus spring viremia virus (SVCV), suggesting that CcNLRP3 might be involved in the immune defense of common carp. The results of co-IP, spot formation, oligomerization and fluorescence localization showed that CcNLRP3 could interact with CcASC and assemble into inflammasome. The cytotoxicity assays showed that CcNLRP3 inflammasome was involved in the pyroptosis induced by CcGSDME. At the same time, CcNLRP3 could directly interact with CcCaspase-A/B and result in increased Caspase-B enzyme activity and LDH release, indicating that CcNLRP3 could also form inflammasome through ASC-independent pathway. Taken together, the results provide targets and theoretical basis for the prevention and control of infectious diseases in aquaculture.

Mitigation of Acute Hepatotoxicity Induced by Cadmium Through Morin: Modulation of Oxidative and Pro-apoptotic Endoplasmic Reticulum Stress and Inflammatory Responses in Rats

Cadmium (Cd) is a toxic heavy metal with significant environmental health hazards. It enters the body through various routes with tissue accumulation. The relatively longer half-life with slow body clearance significantly results in hepatotoxicity during its liver detoxification. Therefore, researchers are exploring the potential use of herbal-derived phytocomponents to mitigate their toxicity. Here, we investigated, for the first time, the possible ameliorative effect of the phytochemical Morin (3,5,7,29,49-pentahydroxyflavone) against acute Cd-induced hepatotoxicity while resolving its underlying cellular mechanisms in a rat animal model. The study involved 50 adult male Sprague–Dawley rats weighing 200–250 g. The animals were divided into five equal groups: control, Cd, Morin100 + Cd, Morin200 + Cd, and Morin200. The 2nd, 3rd, and 4th groups were intraperitoneally treated with Cd (6.5 mg/kg), while the 3rd, 4th, and 5th groups were orally treated with Morin (100 and 200 mg/kg) for 5 consecutive days. On the 6th day, hepatic function (serum ALT, AST, ALP, LDH enzyme activities, and total bilirubin level) testing, transcriptome analysis, and immunohistochemistry were performed to elucidate the ameliorative effect of Morin on hepatotoxicity. In addition to restoring liver function and tissue injury, Morin alleviated Cd-induced hepatic oxidative/endoplasmic reticulum stress in a dose-dependent manner, as revealed by upregulating the expression of antioxidants (SOD, GSH, Gpx, CAT, and Nrf2) and decreasing the expression of ER stress markers. The expression of the proinflammatory mediators (TNF-α, IL-1-β, and IL-6) was also downregulated while improving the anti-inflammatory (IL-10 and IL-4) expression levels. Morin further slowed the apoptotic cascades by deregulating the expression of pro-apoptotic Bax and Caspase 12 markers concomitant with an increase in anti-apoptotic Blc2 mRNA expression. Furthermore, Morin restored Cd-induced tissue damage and markedly suppressed the cytoplasmic expression of JNK and p-PERK immunostained proteins. This study demonstrated the dose-dependent antioxidant hepatoprotective effect of Morin against acute hepatic Cd intoxication. This effect is likely linked with the modulation of upstream p-GRP78/PERK/ATF6 pro-apoptotic oxidative/ER stress and the downstream JNK/BAX/caspase 12 apoptotic signaling pathways.

Association between Oxidative Stress with Psychological and Biochemical Variables in a Sample of Healthy Mexican People: A Cross-Sectional Study.

Oxidative stress (OS) has been linked to cell damage and chronic disease development; however, the study of psychological factors related with OS has been limited, as has its relationship with biochemical and personal variables. Therefore, the aim of this study was to evaluate the association between a wide variety of personal, psychological, and biochemical factors with OS in a sample of healthy Mexican people. A total of 134 participants, from which 70 (52%) were women, without known chronic conditions were included in the study, and the molecule 8-hydroxy-2'-deoxyguanosine (8-OHdG) was also measured as a marker of OS. We observed in the multivariate analysis of the whole sample that depressive symptoms (measured with CES-D scale) were the only psychological variable significantly associated (positively) with 8-OHdG. In addition, the following sociodemographic variables were associated with 8-OHdG: age, schooling (positively correlated), and the frequency of vitamins/antioxidant consumption (negatively correlated). The biochemical variables of erythrocytes in urine and amylase were positively correlated with 8-OHdG, while glucose was negatively correlated with it. Additional biochemical variables were associated in the multivariate analysis of each sex, including the positive correlation of LDL-cholesterol, LDH enzyme, lymphocytes, and the negative correlation of phosphorus and eosinophils in women's samples, as well as the positive correlation of potassium, uric acid, and leucocytes in urine and the negative correlation of erythrocytes and lipase in the men's samples. In conclusion, depression was the only psychological variable positively correlated with 8-OHdG after adjusting for confounders, and new associations with biochemical variables were found with some differences between sexes.

Chiral separation and bioactivities of six pairs of enantiomeric dilignans from Magnolia officinalis var. biloba

Six pairs of enantiomeric dilignans, (+)/(−)-magdiligols A–F, have been isolated from an ethanolic extract of the barks of Magnolia officinalis var. biloba. Their chemical structures were elucidated by extensive spectroscopic analyses, NMR calculation with DP4+ analysis, and the electronic circular dichroism spectra calculation. (+)/(−)-1–3 possessed a dihydrobenzopyran ring, while a propyl chain of 1 was linked via ether bond. (+)/(−)-Magdiligols D and E ((+)/(−)-4 and 5) were dilignans possessing a furan ring. (+)-Magdiligol B ((+)/(−)-2), (+)/(−)-magdiligol C ((+)/(−)-3), and racemes 2, 3, and 5 showed potential hepatoprotective effects against APAP-induced HepG2 cell damage, increased the cell viability from 65.4% to 72.7, 78.7.76.6, 73.9, 77.9 and 73.2%, via decreasing the level of the live enzymes ALH and LDH consistently. (+)/(−)-Magdiligols B–D ((+)/(−)-2–4) and (+)/(−)-magdiligol F ((+)/(−)-6) exhibited significant antioxidative activity. (+)/(−)-Magdiligols B–C ((+)/(−)-2 and 3), (−)-magdiligol D ((−)-4), and (+)-magdiligol E ((+)-5) displayed significant PTP1B inhibitory activity with IC50 values 1.41–3.42 μM. (+)/(−)-Magdiligol B ((+)/(−)-2), and its raceme (2) demonstrated α-glucosidase inhibitory activity with the IC50 values 1.47, 2.88 and 1.85 μM, respectively.