Anti­oxidation effect of Genistein in vascular endothelial cell after H2O2 stress.

Atherosclerosis (AS) is a chronic inflammatory disease characterized by increased oxidative injury in vascular endothelial cells. Inhibiting the oxidative damage of vascular endothelial cells can effectively prevent the occurrence and development of AS. Of note, Genistein (GEN; ID no. 5280961) is phytochemical found in legume family which has flavonoid properties with multiple potential biological activities including antioxidant, anti­inflammatory and anticancer. Antioxidant capacity of GEN has a potential protective effect on vascular endothelial cells after oxidative stress. In the present study, the protective effect of GEN on H2O2­induced oxidation damage was investigated in human vascular endothelial cells (HUVECs). Following GEN pretreatment of HUVECs, H2O2 was added, and apoptosis was detected by flow cytometry, and the expression of relevant genes and proteins was detected by PCR and westerner blot. The results of the present study revealed that GEN significantly enhanced the cell survival rate and decreased the apoptotic rates of HUVECs after H2O2 stress. Besides, GEN reduced the accumulation of intracellular reactive oxygen species by enhancing activity of antioxidant enzymes glutathione peroxidase, superoxide dismutase (SOD) and glutathione peroxidase. Moreover, GEN also inhibited the apoptosis of vascular endothelial cells and enhanced the activation of the nuclear factor erythroid2­related factor 2 (Nrf2)/heme oxygenase­1 (HO­1)/SOD pathway. Collectively, it was identified that GEN is an effective antioxidant which can reduce the oxidative damage by H2O2 through the Nrf2/HO­1/SOD signaling pathway in HUVECs.

A dual donor-acceptor fluorescent probe with viscosity response and lipid droplets targeting to initiate oxidative stress for tumor elimination.

A dual donor-acceptor photosensitizer TCN-2 prepared based on single donor-acceptor could fulfil lipid droplets targeting to trigger apoptosis and tumor growth arrest. Meanwhile, all of experiments both in phosphate buffer solution and intracellular surroundings have demonstrated that TCN-2 catalyzed the production of type I as well as type II reactive oxygen species, forming a hybrid reactive oxygen species pattern, indicating that TCN-2 could be applied to initiate a series of biological responses triggered by oxidative stress within most high-viscosity solid tumors. In addition, TCN-2 also has the capability of fluorescence imaging, which could perfectly combine therapeutic imaging to achieve therapeutic effects while identifying cancerous lesions. Due to the structural design of double electron-absorbing groups, TCN-2 retained excellent lipophilicity while enhancing solubility in the biological environment. Terrific biocompatibility, minimal phototoxic damage to normal cells and tissues, and specific driving to prescriptive organelles to maximize therapeutic effects were used to enhance the therapeutic effect of photodynamic therapy to cease disease progression.

Protein-Protein Interactions: Oxidative Bacterial Two Hybrid.

Protein-protein interaction studies are essential to understand how proteins organize themselves into interaction networks and thus influence cellular processes. Protein binding specificity depends on the correct three-dimensional folding of the polypeptide sequences. One of the forces involved in the structuring and stability of proteins is the formation of disulfide bonds. These covalent bonds are formed posttranscriptionally by the oxidation of a pair of cysteine residues and can serve structural, catalytic, or signaling roles. Here, we describe an engineered E. coli adenylate cyclase mutant strain with an oxidative cytoplasm that promotes correct folding of proteins with disulfide bonds. This genetic background expands the set of host strains suitable for studying protein-protein interactions in vivo by the adenylate cyclase two-hybrid approach.

Toxicological approaches as tool to assess the effects of a mixture of photocatalytic degradation products originated from the unregulated neonicotinoid acetamiprid employing a terrestrial organism (Eisenia andrei).

Acetamiprid (ACT) has been detected in several water sources in Latin America. The presence of its degradation products in the environment is not negligible and transformation products (TPs) significantly contribute to environmental health risks. Although advanced oxidative processes are promising for the treatment of this neocotinoid, effects of these are still unknown. In this context, the effects of a mixture of photocatalytic degradation products resulting from an ACT treatment for 90 min employing TiO2/UV on cytotoxicity and oxidative stress parameters in Eisenia andrei earthworms in acute and chronic experiments using typical Latin American soil were assessed. Acute contact tests were performed (72 h) using a filter paper moistened with an ACT solution and a chronic test was performed using Oxisoil (200 g) moistened with an ACT solution for 45 days. Catalase (CAT) and glutathione-S-transferase (GST) activities, reduced glutathione (GSH) levels and cytotoxicity (cellular eleocyte and amoebocyte assessments) were investigated. Over 75 % of ACT was degraded within the first 15 min of treatment, with levels below the limit of detection after 60 min. The acute test revealed greater cytotoxic effects associated with the effluents treated for T0 and T15 min, with decreased cell density noted after 48 h of exposure, in addition to CAT induction (in all treatments) and GST induction following T0, T15 and T90 min exposures. Concerning the chronic assay, decreases in cell density (T0, T15, T60 and T90 min) and viability (T0, T60 and T90 min) were observed after 45 days, in addition to induced CAT activity following T0, T15 and T60 exposures and GST induction following the T60 min exposure. Reduced glutathione levels were unaltered, comprising the least sensitive biomarker among the investigated parameters to the treated effluent exposures. The mixture of ACT degradation products can cause toxic effects to non-target organisms, despite parent compound degradation, alerting for the need for ecotoxicological tests to prove decreased effluent toxicity, in addition to the improvement of degradation techniques.

From genes to systems: The role of food supplementation in the regulation of sepsis-induced inflammation.

Systemic inflammation includes a widespread immune response to a harmful stimulus that results in extensive systemic damage. One common example of systemic inflammation is sepsis, which is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection. Under the pro-inflammatory environment of sepsis, oxidative stress contributes to tissue damage due to dysfunctional microcirculation that progressively causes the failure of multiple organs that ultimately triggers death. To address the underlying inflammatory condition in critically ill patients, progress has been made to assess the beneficial effects of dietary supplements, which include polyphenols, amino acids, fatty acids, vitamins, and minerals that are recognized for their immuno-modulating, anticoagulating, and analgesic properties. Therefore, we aimed to review and discuss the contribution of food-derived supplementation in the regulation of inflammation from gene expression to physiological responses and summarize the precedented potential of current therapeutic approaches during systemic inflammation.

Dapagliflozin and metformin in combination ameliorates diabetic nephropathy by suppressing oxidative stress, inflammation, and apoptosis and activating autophagy in diabetic rats.

Considering the effects of sodium-glucose cotransporter inhibitors and metformin on the kidneys, a combination of both agents is postulated to provide protection against diabetic nephropathy (DN). We examined the potential protective effects of dapagliflozin, metformin, and their combination on kidney injury in rats with type 2 diabetes. Diabetic (DM) rats were administered dapagliflozin (1.0 mg/kg/day), metformin (100 mg/kg/day), or a combination (dapagliflozin 0.5 mg/kg/day plus metformin 50 mg/kg/day) by oral gavage for 4 weeks. Dapagliflozin monotherapy or in combination with metformin was more effective than metformin monotherapy in attenuating renal dysfunction, improving renal organic anion transporter 3 expression, and activating renal autophagy by modulating the AMPK/mTOR/SIRT1 axis in DM rats. Interestingly, dapagliflozin monotherapy exhibited greater efficacy in suppressing renal oxidative stress in DM rats than metformin or the combination treatment. Renal and pancreatic injury scores decreased in all treatment groups. Apoptotic markers were predominantly reduced in dapagliflozin monotherapy and combination treatment groups. The low-dose combination treatment, through synergistic coordination, appeared to modulate oxidative, autophagic, and apoptotic signaling and confer significant renoprotective effects against DM-induced complications. In addition, a low dose of the combination might be beneficial to patients by avoiding the risk of side effects of the medication. Future clinical trials are necessary to study the nephroprotective effects of the combined treatment at a low dosage in patients with diabetes.

Pioglitazone ameliorates DOX-induced cognitive impairment by mitigating inflammation, oxidative stress, and apoptosis of hippocampal neurons in rats.

Doxorubicin (DOX) is broadly used as a medication for cancer treatment. However, DOX has been connected with chemotherapy-related complications, for instance, cognitive impairment (chemobrain). Chemobrain developed in up to 70% of cancer patients; therapeutic is unavailable. This study investigated the preventive effect of pioglitazone (PIO) on neurotoxicity caused by (DOX) in the hippocampus. Forty rats were separated into four groups; control (normal saline 10 ml/kg), DOX (5 mg/kg, intraperitoneally every 3rd day, equivalent to 20 mg/kg cumulative dose), PIO (2 mg/kg in drinking water), and DOX+PIO (DOX, 5 mg/kg, intraperitoneally every 3rd day concurrently PIO, 2 mg/kg in drinking water) and duration of drug treatment lasted for 14 days. The animals were subjected to contextual fear memory tests to characterize the cognitive impairment following DOX treatment. ELISA assessed hippocampal protein expression related to inflammation, oxidative damage, and apoptosis. DOX-treatment produced significant reduction in freezing duration in contextual fear memory tests, which was reversed by PIO co-administration. DOX increased neuroinflammation, oxidative stress, apoptosis, and mitochondrial activity by increasing NF-κB and COX-2 levels, reducing SOD levels, and increasing Bax, caspase-3, and lipid peroxidation. However, DOX did not affect GSH or catalase levels. PIO co-administration reduces NF-κB, COX-2, MDA, Bax, and caspase-3 levels and improves mitochondrial activity and SOD expression. To sum up, DOX therapy accelerates cognitive decline in rats by increasing neuroinflammation, oxidative stress, mitochondrial dysfunction, lipid peroxidation, and apoptosis. PIO is a promising treatment for DOX-induced cognitive impairment.

Ginsenoside RK1 improves cognitive impairments and pathological changes in Alzheimer's disease via stimulation of the AMPK/Nrf2 signaling pathway.

BACKGROUND: The pathogenesis of Alzheimer's disease (AD) is complex, resulting in unsatisfactory effects of single-target therapeutic drugs. Accumulation evidence suggests that low toxicity multi-target drugs may play effective roles in AD. Ginseng is the root and rhizome of Panax ginseng Meyer, which can be used not only as herbal medicine but also as a functional food to support body functions. Ginsenoside RK1 (RK1), obtained from ginseng plants through high-temperature treatment, has antiapoptotic, antioxidant, anti-inflammatory effects and these events are involved in the development of AD. So, we believe that RK1 may be an effective drug for the treatment of AD. HYPOTHESIS/PURPOSE: We aimed to investigate the potential protective effects and mechanisms of RK1 in AD. METHODS: Neuronal damage was detected by MTT assay, LDH assay, immunofluorescence and western blotting. Oxidative stress was measured by JC-1 staining, reactive oxygen species (ROS) staining, superoxide dismutase (SOD) and malonaldehyde (MDA). The cognitive deficit was measured through morris water maze (MWM) and novel object recognition (NOR) tests. RESULTS: RK1 attenuated Aß-induced apoptosis, restored mitochondrial membrane potential (ΔΨm), and reduced intracellular levels of ROS in both PC12 cells and primary cultured neurons. In vivo, RK1 significantly improved cognitive deficits and mitigated AD-like pathological features. Notably, RK1 demonstrated superior efficacy compared to the positive control drug, donepezil. Mechanistically, our study elucidates that RK1 modulates the phosphorylation of AMP-activated protein kinase (AMPK) and its downstream target, NF-E2-related factor 2 (Nrf2), leading to the optimization of mitochondrial membrane potential, reduction of ROS levels, and mitigation of AD-like pathology. It's noteworthy that blocking the AMPK signaling pathway attenuated the protective effects of RK1. CONCLUSION: RK1 demonstrates superior efficacy in alleviating cognitive deficits and mitigating pathological changes compared to donepezil. These findings suggest the potential utility of RK1-based therapies in the development of treatments for AD.

Upregulation of mitochondrial telomerase reverse transcriptase mediates the preventive effect of physical exercise on pathological cardiac hypertrophy via improving mitochondrial function and inhibiting oxidative stress.

Physical exercise is a non-pharmacological intervention that helps prevent pathological cardiac hypertrophy. However, the underlying molecular mechanisms remain unclear. Telomerase reverse transcriptase (TERT) has non-telomeric functions such as protection against mitochondrial dysfunction and oxidative stress, and its myocardial expression is upregulated by physical exercise. Here, we found that physical exercise caused myocardial upregulation of mitochondrial TERT and sustenance during transverse aortic constriction (TAC)-induced cardiac hypertrophy. Overexpression of mitochondrial-targeted TERT (mito-TERT) via adeno-associated virus serotype 9 carrying the TERT-coding sequence fused with N-terminal mitochondrial-targeting sequence improved cardiac function and attenuated cardiac hypertrophy. Mechanistically, mito-TERT ameliorated mitochondrial dysfunction and oxidative stress, which were associated with improving the activity and subunit composition of complex I. Remarkably, the telomerase activator TA-65 also exhibited an antihypertrophic effect. Collectively, our results reveal a significant role for mito-TERT in mediating the antihypertrophic effect of physical exercise and demonstrate that TERT is a potential drug target for treating cardiac hypertrophy.

Ameliorative effect of scopolamine-induced cognitive dysfunction by Fufangmuniziqi formula: The roles of alkaloids, saponins, and flavonoids.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufangmuniziqi formula (FFMN), a traditional Uyghur medicine used in China, is derived from an ancient Uyghur medical book and consists of 13 herbs. The herbs of FFMN, such as Peganum harmala L., Glycyrrhiza uralensis Fisch., and Nigella glandulifera, have been demonstrated to have acetylcholinesterase (AChE) inhibitory, anti-neuroinflammatory, or antioxidant effects. Therefore, FFMN may have a good anti-Alzheimer's disease (AD) effect, but its specific action and mechanism need to be further proven. AIM OF THE STUDY: This study aims to investigate the anti-AD effects of FFMN and the role played by alkaloids, flavonoids, and saponins in anti-AD. MATERIALS AND METHODS: The alkaloids, flavonoids, and saponins fractions of FFMN were prepared by macroporous resin chromatography. The absorbed ingredients in the drug-containing serum were identified by UPLC⁃Q⁃TOF⁃MS. An AD mouse model was established by intraperitoneal injection of scopolamine (SCO). The role of different fractions of FFMN in the anti-AD process was examined by Morris water maze (MWM), in-vitro cell, and AChE inhibition assay. RESULTS: A total of 20 ingredients were identified in the serum samples collected after oral administration of FFMN, and seven compounds were selected as candidate active compounds. MWM experiments showed that different fractions of FFMN could significantly improve SCO-induced learning memory impairment in mice. The alkaloids fraction (ALK) regulated cholinergic function by inhibiting AChE activity, activating choline acetyltransferase activity, and protein expression. Flavonoids and saponins were more potent than the ALK in downregulating pro-inflammatory factors or inflammatory mediators, such as TNF-α, MPO, and nitric oxide. Western blot results further confirmed that flavonoids and saponins attenuated neuroinflammation by inhibiting the phosphorylation of IκB and NF-κB p65. This result was also verified by in-vitro cellular assays. FFMN enhanced antioxidant defense by increasing the activity of superoxide dismutase and reducing the production of MDA. Combined with cellular experiments, flavonoids and saponins were proven more protective against oxidative damage. CONCLUSION: FFMN improved cognitive and memory impairment in the SCO-induced AD mouse model. ALK mainly enhanced the function of the cholinergic system. Flavonoid and saponin fractions mainly attenuated neuroinflammation and oxidative stress by modulating the NF-κB pathway. All these findings strongly suggested that the combination of alkaloid, flavonoid, and saponin fractions derived from FFMN is a promising anti-AD agent that deserves further development.

Danggui Shaoyao San: Chemical characterization and inhibition of oxidative stress and inflammation to treat CCl4-induced hepatic fibrosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Danggui Shaoyao San (DSS) has effective in treating hepatic ascites and liver disease. AIM OF THE STUDY: To explore the chemical characterization of DSS and protective effect on CCl4-induced hepatic fibrosis and its mechanism, especially its anti-oxidative stress and anti-inflammation. MATERIALS AND METHODS: The chemical characterization of DSS was determined by HPLC-Q-Exactive Orbitrap MS. And the antioxidant activity of DSS in vitro was determined. The hepatic fibrosis model was established using intragastric administration of 40% CCl4/soybean oil (v/v) twice weekly for 13 weeks. From 6th week, the DSS group and the positive control group were given DSS (2, 4, 8 g/kg/d) and silymarin (50 mg/kg/d), respectively. The livers of rats were examined histologically by H&E. The ALT, AST, ALB and TBIL were determined, and hepatic fibrosis markers (HA, LN, CIV, PIIINP), oxidative stress (SOD, MDA, GST, GSH) and inflammatory factor (IL-6, TNF-α) were tested using ELISA kits. In addition, the levels of TAC, TOS, LOOH and AOPP in the liver were also determined. RESULTS: The chemical characterization of DSS was determined by HPLC-Q-Exactive Orbitrap MS. The results show that DSS mainly includes triterpenoids, monoterpenes, phenols, sesquiterpenes, butyl phthalide, etc., and DSS has good antioxidant activity in vitro. In addition, the ALT, AST and TBIL of rats were remarkably reduced after treatment with DSS at three doses. Liver histopathological analysis showed that DSS alleviated the inflammatory infiltration, hepatocyte swelling, necrosis and hepatic fibrosis induced by CCl4. DSS significantly decreased HA, IV-C, PIIINP and LN. Further determination showed that DSS significantly increased TAC, OSI and decreased TOC, LOOH and MDA, indicating that DSS could regulate redox balance and reduce lipid peroxidation in vivo. DSS also increased the activity of GST, SOD and GSH concentration. In addition, DSS also reduced IL-6 and TNF-α. CONCLUSIONS: In this study, we described the chemical characterization of DSS and found that it has good antioxidant activity. We proved that DSS has the functions of reducing oxidative stress, anti-inflammatory, protecting liver cells and reducing hepatic fibrosis.

Ameliorative effect of traditional polyherbal formulation on TNF-α, IL-1ß and Caspase-3 expression in kidneys of wistar rats against sodium fluoride induced oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Sharbat-e-bazoori Motadil (SBM) is a polyherbal formulation that have been used for centuries as a part of the Unani system of medicine for renal disease. AIM OF THE STUDY: The objective of this study was to explore and validate the nephroprotective potential of sugar-free SBM (SF-SBM) and its mechanisms of action against sodium fluoride (NaF)-induced nephrotoxicity in HEK-293 cells. Additionally, the study aimed to assess the quality control of SF-SBM and investigate its effects using an in vivo rat model with pattern recognition following oral administration of SF-SBM. MATERIALS AND METHODS: The nephroprotective effect of SF-SBM was investigated using both an HEK-293 cell line and Wistar rats. Nephrotoxicity was induced in these models by administering NaF at a concentration of 600 ppm (parts per million) for a duration of seven days. The SF-SBM formulation was standardized using high-performance thin-layer chromatography (HPTLC) to assess the presence of marker compounds, namely gallic acid, quercetin, and ferulic acid. Metabolite characterization of SF-SBM was carried out using ultra-high-performance liquid chromatography mass spectrometry (UPLC-MS) with a monolithic capillary silica-based C18 column. This analytical technique allowed for the identification of bioactive substances and verification of the identified markers. Acute toxicity of SF-SBM was evaluated in Wistar rats by administering a single oral dose of 2000 mg/kg of SF-SBM. The nephroprotective efficacy of SF-SBM was further assessed at low (LD), medium (MD) and high (HD) doses of 32.1, 64.2, and 128.4 mg/kg, respectively, administered orally. Nephrotoxicity was induced in Wistar rats by adding NaF to their drinking water for seven days. Biochemical and urine markers were analyzed to evaluate the antioxidant, inflammatory, and apoptotic potential of SF-SBM. Additionally, histopathological analysis and immunohistochemical alterations in the expression of caspase-3 and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-4 (NOX-4) in kidney tissue were performed to confirm the findings of the in vivo experiments. Furthermore, in vivo pattern recognition of SF-SBM metabolites, identified through GC-MS metabolomics, and in-silico docking analysis of major metabolites in plasma were conducted to gain further insights. RESULT: Phytochemical analysis using HPTLC, TLC-bioautography, and UPLC-MS revealed the presence of several bioactive constituents in SF-SBM, including ferulic acid, gallic acid (GA), ellagic acid, quercetin, and apigenin. These compounds exhibit diverse pharmacological properties. In vitro studies demonstrated the protective effect of SF-SBM on HEK-293 cell line against nephrotoxicity. The acute toxicity study of SF-SBM at a dose of 2000 mg/kg showed no mortality or signs of toxicity throughout the 14-day observation period. In the in vivo studies, administration of NaF resulted in significant elevation (P < 0.001) of biochemical and urine parameters, indicating oxidative, inflammatory, and apoptotic stress. Histopathological examination revealed severe depletion of Bowman's capsule, and immunohistochemistry demonstrated negative immunostaining for caspase-3 and reduced NOX-4 reactions. Pre-treatment with SF-SBM significantly attenuated the elevated biochemical and urine markers, restored the antioxidant enzyme levels (such as SOD, CAT, GSH, GPx and NO), and regulated the expression of inflammatory cytokines (TNF-α, IL-1ß, CASP-3) in kidney tissue at doses of SF-SBM-MD (64.2 mg/kg) and SF-SBM-HD (128.4 mg/kg), showing comparable results to those of α-Ketoanalogue. Histopathological assessment demonstrated improvements in tissue damage. Pattern recognition analysis of SF-SBM identified the presence of 56 metabolites at different time intervals. Additionally, in-silico studies revealed strong interactions of SF-SBM with a binding energy of -6.5 and -5.6 kcal for 4C2N. CONCLUSION: The phytoconstituents present in SF-SBM play a crucial role in its nephroprotective action by acting as potent antioxidants and reducing proinflammatory and apoptotic damage in rat cells. This indicates that SF-SBM has promising potential for the treatment of nephrotoxicity.

Geniposide protects against neurotoxicity in mouse models of rotenone-induced Parkinson's disease involving the mTOR and Nrf2 pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Fructus Gardeniae, with the effects of discharging fire, eliminating vexation, reducing fever and causing diuresis, and cooling blood to remove apthogentic heat, could be used to treat Parkinson's disease (PD). Geniposide, as the main active ingredient of Fructus Gardeniae, has been shown to have neuroprotective effects in several rodent models. Rotenone, a commonly used neurotoxin, induced PD model progresses slowly, but simulates the pathological changes of PD's slow progression. AIM OF THE STUDY: Herein, we mainly investigated the neuroprotective effects of geniposide on rotenone-induced mouse model of PD and the underlined mechanism. MATERIALS AND METHODS: C57BL/6 mice were treated with rotenone (30 mg/kg, p. o.) daily for 60 days. Geniposide (25 and 50 mg/kg, p. o.) were administered at alterative day 30 min before rotenone. On day 60, the challenging beam, spontaneous activity, and adhesive removal tests were performed to evaluate the motor activity. Dopamine, DOPAC and HVA levels were detected by UPLC-MS/MS methods. Dopaminergic neurodegeneration was assessed using immunohistochemistry staining. ROS production, MDA level and GSH: GSSG ratio were measured to analyze oxidative stress. Cleavage of PARP and caspase-3 were detected to assess neuronal apoptosis. The expression of Nrf2 and mTOR signaling were detected using Western blot. RESULTS: Geniposide improved motor dysfunction, restored neurotransmitters levels, and attenuated dopaminergic neurodegeneration induced by rotenone in mice. Geniposide suppressed rotenone-induced neuronal oxidative damage associated with Nrf2 signaling, and neuronal apoptosis involving mTOR pathway. CONCLUSIONS: Geniposide may exert a neuroprotective effect in a mouse model of PD by rotenone, and this effect might be relevant to Nrf2 associated antioxidant signaling and mTOR involved anti-apoptosis pathway.

Neuroprotective effects of Petiveria alliacea on scopolamine-induced learning and memory impairment mouse model.

ETHNOPHARMACOLOGY RELEVANCE: Petiveria alliacea L., commonly known as macura and gully root, is an important medicinal plant used in the Caribbean and Central America to treat ailments associated to the central nervous system, including poor memory. AIM OF THE STUDY: To assess the effects of the P. alliacea leaves methanol fraction (PMF) on a scopolamine-induced learning and memory impairment mouse model related to acetylcholinesterase activity and oxidative stress. MATERIAL AND METHODS: After PMF administration at doses of 500 or 900 mg/kg, cognitive ability was evaluated using the Morris water maze (MWM), Y-maze (YM) and novel object recognition (NOR) tests. The mouse brain tissue was further assessed for acetylcholinesterase activity and antioxidant activity. Levels of oxidative stress were also evaluated by measuring malondialdehyde (MDA) and glutathione activity. Acute toxicity was also evaluated. RESULTS: PMF led to memory improvement in the behavioral tests in mice with scopolamine-induced cognitive impairment. Moreover, PMF inhibited acetylcholinesterase activity and showed antioxidant potential that in turn attenuated cholinergic degradation. Additionally, PMF increased glutathione levels and glutathione reductase activity and reduced MDA levels in the brain. Moreover, no acute toxicity was detected with the use of PMF. CONCLUSION: In a mouse model of scopolamine-induced cognitive deficit, PMF exhibited protective effects, decreasing oxidative damage and regulating cholinergic function in the brain bearing significant memory enhancing potency. These data suggest that PMF is a promising candidate for developing therapies for neurodegenerative disorders.

Protective effects of scutellaria-coptis herb couple against non-alcoholic steatohepatitis via activating NRF2 and FXR pathways in vivo and in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Scutellaria-coptis herb couple (SC) is a classic herbal pair used in many Traditional Chinese Medicine (TCM) formulations in the treatment of endocrine and metabolic deseases. Diabetes mellitus and non-alcoholic steatohepatitis (NASH) are both endocrine and metabolic diseases. Previous studies have shown that SC has anti-diabetic effects. However, the effect and mechanism of SC against NASH remains unclear. AIM OF THE STUDY: This study aimed to demonstrate the effect and mechanism of SC against NASH through the nuclear factor-erythroid 2-related factor 2 (Nrf2) and farnesoid X receptor (FXR) dual signaling pathways in vivo and in vitro. MATERIALS AND METHODS: The high fat diet-fed rat model, and HepG2 and RAW264.7 cell models were used. Serum biochemical indexes and liver histopathological changes were examined. Metabolomics, transcriptomics, and flow cytometry were performed. RT-qPCR and western blot analysis were performed to provide expression of NRF2 and FXR pathway signal molecules during SC's anti-NASH treatment in vivo and in vitro. RESULTS: SC had anti-NASH effects in vivo with significantly improvement of serum NASH biochemical index and hepatopathological structure; meanwhile, SC significantly elevated the expression levels of FXR protein in liver and intestinal tissues, and cholesterol 7a-hydroxylase (CYP7A1) protein in liver. The mRNA expression levels of Takeda G protein receptor 5 (TGR5), CYP7A1, fibroblast growth factor receptor-4 (FGFR4), FXR, small heterodimer partner (SHP), fibroblast growth factor 15/19 (FGF15/19) and glucagon-like peptide-1 (GLP-1) were significantly elevated by SC. SC reduced the levels of NorCA, isoLCA and α-MCA in the feces of NAFLD rats. In vitro, SC-containing serum (SC-CS) was found to significantly reduce intracellular lipid deposition, inhibit ROS production, reduce intracellular Malondialdehyde (MDA) and IL-1ß levels, and enhance the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Six differential genes closely related to oxidative stress and Nrf2 were identified by transcriptomic analysis. SC-CS up-regulated the expression of NRF2, and reduced the expression of TXNIP and Caspase-1 genes in RAW264.7 cells. In addition, SC-CS reduced the expression of Keap1 and NF-κB, and up-regulated the expression of Nrf2, heme oxygenase-1 (HO-1), quinone oxidoreductase 1 (NQO1), and SOD; SC-CS elevated the protein level of NRF2, and reduced the protein level of TXNIP in HepG2 cells. CONCLUSIONS: the mechanisms of SC action against NASH was closely related to the simultaneous activations of both NRF2 and FXR signaling pathways. These findings provide a new insight into the anti-NASH application of SC in clinical settings and demonstrate the potential of SC in the treatment of NASH.

"Yajieshaba" prevents acute alcoholic liver injury and repairs the intestinal mucosal barrier.

ETHNOPHARMACOLOGICAL RELEVANCE: An essential factor related to the acute alcoholic liver injury is damage to the intestinal mucosal barrier. Yajieshaba (YJSB) is a commonly used formulation of Dai people in China and protects the liver. AIM OF THE STUDY: This study investigated whether YJSB can prevent acute alcoholic liver injury by regulating the intestinal mucosal barrier. MATERIALS AND METHODS: The mice received 0.39 g/kg, 1.17 g/kg, and 3.51 g/kg dose YJSB for 7 days, a mouse model of acute alcoholic liver injury was established by a single instillation of 56% alcohol. Plasma biochemical markers were analyzed, liver injury was identified by histopathology, lipopolysaccharide (LPS), nuclear factor-k-gene binding (NF-κB), hepatic inflammatory factors, oxidative stress factors and reactive oxygen species (ROS) content was analyzed. The morphological changes of intestinal histology were observed by H&E staining, and the ultrastructure of ileal cells was observed by transmission electron microscopy. Immunofluorescence and Western blot was used to determine the expression levels of transporters and enzymes involved in Claudin 1, Occludin and zona occludens 1 (ZO-1) homeostasis in the liver and intestine. RESULTS: The findings showed that YJSB reduced the levels of aspartate aminotr ansferase (AST), alanine aminotransferase (ALT) and total bile acid (TBA), both of which are indicators of liver function and had a protective effect against liver injury. In the liver homogenate, YJSB reduced the level of LPS, NF-κB, tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6), decreased the level of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-PX) and catalase (CAT) and ROS. The results of hematoxylin and eosin (H&E) staining and transmission electron microscopy analysis revealed that YJSB reduced the degree of damage to intestinal tissue and intracellular organelles, implying that YJSB can reduce the "attack factor" that causes intestinal barrier damage, increase the "defense factor" that protects the intestinal barrier. The results of immunohistochemistry and Western blotting analysis showed that YJSB could increase the expression of claudin 1, occludin, and ZO-1 proteins, suggesting that the mechanism of action of YJSB against acute alcohol liver injury involves the upregulation of the expression of the intestinal barrier-related proteins and the repair of the damaged intestinal barrier. CONCLUSIONS: YJSB can block LPS, oxidative stress factors, and other harmful substances in the blood and protect the liver resisting acute alcoholic liver injury.

Synthetic azo-dye, Tartrazine induces neurodevelopmental toxicity via mitochondria-mediated apoptosis in zebrafish embryos.

Tartrazine (TZ), or E 102 or C Yellow, is a commonly used azo dye in the food and dyeing industries. Its excessive usage beyond permissible levels threatens human health and the aquatic environment. While previous studies have reported adverse effects such as mutagenicity, carcinogenicity, and reproductive toxicity. Our study aimed to comprehensively evaluate the developmental neurotoxicity of TZ exposure via biochemical and behavioral examinations and explored the underlying mechanism via gene expression analyses. TZ at an environmentally relevant concentration (50 mg/L) significantly induces oxidative stress, altered antioxidant (SOD, CAT and GSH) response, triggered cellular damage (MDA and LDH), and induced neuro-biochemical changes (AChE and NO). Gene expression analyses revealed broad disruptions in genes associated with antioxidant defense (sod1, cat, and gstp1), mitochondrial dysfunction (mfn2, opa1, and fis1),evoked inflammatory response (nfkb, tnfa, and il1b), apoptosis activation (bcl2, bax, and p53), and neural development (bdnf, mbp, and syn2a). Behavioral analysis indicated altered thigmotaxis, touch response, and locomotion depending on the concentration of TZ exposure. Remarkably, the observed effective concentrations were consistent with the permitted levels in food products, highlighting the neurodevelopmental effects of TZ at environmentally relevant concentrations. These findings provide valuable insights into the underlying molecular mechanisms, particularly the role of mitochondria-mediated apoptosis, contributing to TZ-induced neurodevelopmental disorders in vivo.

Toxic effects and potential mechanisms of zinc pyrithione (ZPT) exposure on sperm and testicular injury in zebrafish.

Zinc pyrithione (ZPT) is widely recognized for its beneficial properties as an antifouling, antibacterial, and antifungal agent. Despite its positive industrial contributions, ZPT has been proven to exhibit toxicity towards various ecosystems, particularly affecting marine life. However, there is still a dearth of comprehensive research on ZPT toxicity and its toxicological mechanism in reproductive systems of aquatic organisms. In our study, we conducted a thorough analysis and unveiled a multitude of abnormalities in zebrafish sperm and testicular tissue caused by ZPT exposure, including a dose-dependent diminishing of testosterone levels, various sperm deformities, decreased sperm concentration and motility, and ROS-induced testicular tissue DNA damage. In addition, our study suggested that ZPT-induced testicular damage is associated with heightened oxidative stress, apoptosis, and possible hyperpolarization of the mitochondrial membrane. Through RNA-seq analysis, a total of 409 DEGs associated with ZPT-induced testicular injury were identified, and the hub gene was determined using a protein-protein interaction network (PPI). The genes and pathways uncovered in this study point to potential mechanisms of ZPT exposure on sperm and testicular injury in zebrafish.

Exploring the mechanism of exopolysaccharides in mitigating cadmium toxicity in rice through analyzing the changes of antioxidant system.

Recently, exopolysaccharides (EPS) were found to alleviate cadmium (Cd) toxicity to crops by regulating the antioxidant system, but the mechanism remains unclear. Herein, by quantitative and transcriptomic approaches, a systematical map of the changes in the antioxidant system was drawn to dissected the underlying mechanism. The results demonstrated that the ascorbate-glutathione cycle (ASA-GSH cycle) is a major contributor. Specifically, compared to the control, the rice exposed to Cd exhibited a significant increase in the GSH pool (about 9-fold at 7 d), but a continuous decrease in the ASA pool (only 15.42% remained at 15 d) and an excessive accumulation of reactive oxygen species (ROS). Interestingly, with the addition of EPS, the increase of the GSH pool significantly slowed down (decreased by 180.18% at 7 d, compared to the Cd-stressed treatment), and the ASA pool remained high (consistently above 70.00% of the control group). ROS also maintained at a good level. Moreover, the activities of enzymatic antioxidants showed the similar trend. By RNA-Seq analysis, multiple genes enriched in ASA-GSH related pathway were screened (such as OsRBOHB, OsGST, OsPOD) for further study. This study provides a foundation for EPS application in agriculture, which also establishes a better way for analyzing antioxidant system.

Chlorophytum borivilianum aqueous root extract prevents deterioration of testicular function in mice and preserves human sperm function in hydrogen peroxide (H2O2)-induced oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Chlorophytum borivilianum (C. borivilianum) (CB) has traditionally been used to treat male sexual dysfunctions and has been claimed to possess aphrodisiac properties. AIM OF THE STUDY: To investigate the ability of CB to ameliorate H2O2-induced oxidative stress in testes and sperm in mice and prevent H2O2-induced oxidative in human sperm. MATERIALS AND METHODS: Oxidative stress was induced in male mice by pre-exposure to 2% H2O2 orally for seven consecutive days, followed by 100 and 200 mg/kg b. w. administration. CB for another seven days. At the end of treatment, mice were sacrificed and testes and epididymal sperm were harvested. Serum FSH, LH and testosterone levels were measured and sperm parameters were obtained. Meanwhile, oxidative stress levels in mice testes and sperm, steroidogenesis and spermatogenesis markers in mice testes were assessed by molecular biological techniques. In another experiment, sperm from thirty-two healthy fertile men were incubated with 200 µM H2O2 and CB (100 and 200 µg/ml) simultaneously and were then evaluated for sperm parameter changes. RESULTS: In mice, CB administration ameliorates persistent increases in oxidative stress and decreases in anti-oxidative enzyme levels in testes and sperm following H2O2 pre-exposure. Additionally, CB also helps to ameliorate deterioration in sperm parameters and testicular steroidogenesis and spermatogenesis and restores the serum FSH, LH and testosterone levels near normal in mice. In humans, CB helps to prevent deterioration in sperm parameters following H2O2 exposure. CONCLUSION: CB is potentially useful to preserve the male reproductive capability and subsequently male fertility in high oxidative stress conditions.