Shenrong Guben Huanshao Pill enhanced stress resistance and extended the lifespan of Caenorhabditis elegans by activating the oxidative stress system via the DAF-16/FOXO signaling pathway.

Introduction: As a traditional medicine, the aerial parts of Leonurus japonicus Houtt. (Lamiaceae), aka motherwort, have been extensively used to treat gynecological diseases. The current study was designed to investigate the longevity properties of the methanolic extract of L. japonicus (MLJ) using Caenorhabditis elegans model system. Methods: The longevity effect of MLJ was determined by lifespan assay. Lipofuscin accumulation, thermotolerance, and body movement were measured to test the effects on the healthspan. The antioxidant capacity of MLJ was investigated by analyzing antioxidant enzyme activities, intracellular reactive oxygen species (ROS) levels, and the survival rate against oxidative stress conditions. Pharyngeal pumping rate and body length were observed to determine the effect of MLJ on aging-related factors. Transcriptional activity of daf-16 was observed under fluorescence microscopy using a transgenic mutant carrying DAF-16::GFP transgene. Results: MLJ could significantly prolong the median and maximum lifespan of worms. In addition, MLJ reduced the accumulation of lipofuscin in aged worms and delayed the age-dependent decrease in locomotion and thermotolerance suggesting its beneficial role in the healthspan. Also, MLJ increased the stress resistance of worms against oxidative stress and decreased intracellular ROS generation by up-regulating the activities of antioxidant enzymes. Additional genetic studies showed that MLJ failed to prolong the lifespan of worms lacking daf-2, age-1, daf-16, and sir-2.1 genes. Moreover, in the presence of MLJ, the nuclear translocation of daf-16 was significantly increased. Conclusion: Collectively, our results demonstrate that the anti-aging properties of MLJ might be attributed to sir-2.1 and insulin/IGF signaling-dependent daf-16/FOXO activation.

As a significant global issue, aging is prompting people's interest in the potential anti-aging properties of Anoectochilus roxburghii (A. roxburghii), a plant traditionally utilized in various Asian countries for its purported benefits in treating diabetes and combating aging. However, the specific anti-aging components and mechanisms of A. roxburghii remain unclear. This study aims to investigate the anti-aging effects and mechanisms of A. roxburghii extract E (ARE). Caenorhabditis elegans (C. elegans) were exposed to media containing different concentrations of ARE whose superior in vitro radical scavenging capacity was thus identified. Lifespan assays, stress resistance tests, and RT-qPCR analyses were conducted to evaluate anti-aging efficacy, reactive oxygen species (ROS) levels, antioxidant enzyme activity, and daf-16, sod-3, and gst-4 levels. Additionally, transcriptomic and metabolomic analyses were performed to elucidate the potential anti-aging mechanisms of ARE. Fluorescence protein assays and gene knockout experiments were employed to validate the impacts of ARE on anti-aging mechanisms. Our results revealed that ARE not only prolonged the lifespan of C. elegans but also mitigated ROS and lipofuscin accumulation, and boosted resistance to UV and heat stress. Furthermore, ARE modulated the expression of pivotal anti-aging genes including daf-16, sod-3, and gst-4, facilitating the nuclear translocation of DAF-16. Significantly, ARE failed to extend the lifespan of daf-16-deficient C. elegans (CF1038), indicating its dependency on the daf-16/FoxO signaling pathway. These results underscored the effectiveness of ARE as a natural agent for enhancing longevity and stress resilience to C. elegans, potentially to human.

ObjectivesIt is well known that intensification of steroid hormones and lipids peroxidation reflect on the membrane of the lysosomes. Isolated lysosomal enzymes in a pathological condition, create the radicals and...

Chronic renal failure (CRF) is usually associated with chronic diseases such as congestive heart failure and diabetes mellitus, the prevalence of which is increased with age. This study is designed to investigate the role of long intergenic non-coding RNA (lincRNA) LINC00963 in renal interstitial fibrosis (RIF) and oxidative stress (OS) of CRF via the forkhead box O (FoxO) signaling pathway. Microarray data and annotated probe files related to CRF were downloaded by retrieving Gene Expression Omnibus (GEO) database to screen differentially expressed lncRNA. Multi Experiment Matrix (MEM) website and dual-luciferase reporter gene assay were used to predict and verify the target gene of LINC00963, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis to identify the major signaling pathways involved. A total of 60 Wistar male rats were randomly selected and divided into the sham (n = 10) and model (n = 50) groups. Five rats in the sham group and thirty rats in the model group were sub-categorized into the control, blank, negative control (NC), LINC00963 vector, si-LINC00963, si-FoxO3, and si-LINC00963 + si-FoxO3 groups (n = 5). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were performed to evaluate the expressions of LINC00963, FoxO3a, TGF-β1, FN, GSH-PX, Bax, and Bcl-2. Measurement of changes in OS indexes including BUN, MDA, GSH-Px, SOD, and Na+-K+-ATP were conducted. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of inflammatory factors including TNF-α, IL-6, ICAM-1 and FN. TUNEL staining was performed to evaluate cell apoptosis. LINC00963 was highly expressed in CRF rats and FoxO3 was predicted and then verified as a target gene of LINC00963. FoxO3 gene participated in the FOXO signaling pathway. Compared with the blank and NC groups, there were significantly decreased expressions of LINC00963, TGF-β1, FN, and Bax in the si-LINC00963 group, while increased expressions of GSH-PX, FoxO3a, and Bcl-2. The vitality values of BUN and MDA in the si-LINC00963 group declined, while enzymatic activities of GSH-Px, SOD and Na+-K+-ATP elevated in comparison to the blank and NC groups. The levels of TNF-α, IL-6, ICAM-1 and FN, and cell apoptosis rate in the si-LINC00963 group decreased in comparison to the blank and NC groups. All the results in the si-LINC00963 group were opposite in the LINC00963 vector and si-FoxO3 groups. Taken together, we conclude that down-regulation of LINC00963 suppresses RIF and OS of CRF by activating the FoxO signaling pathway.

Introduction: There is a complex interaction between the hypothalamus, pituitary, and ovary in the human menstrual cycle. There is increasing evidence about the role of oxidative stress on female reproductive tract. Methods: The purpose of this study was to determine the activities of antioxidant enzymes; superoxide dismutase, glutathione peroxidase, catalase and total antioxidant capacity during the menses, follicular and luteal phases of the menstrual cycle in twenty women. In addition, the relationship between the activity of antioxidant enzyme and estradiol, progesterone, LH, FSH, and testosterone were evaluated. Results: There was no significant difference between the activity of superoxide dismutase, glutathione peroxidase, catalase and total antioxidant capacity during the menses, follicular and luteal phases of the menstrual cycle. We found a significant correlation between LH and FSH with the activity of superoxide dismutase and glutathione peroxidase during the luteal phase (p<0.05). Conclusion: Our results showed that there were not significant differences in activity of antioxidant enzymes during the menstrual cycle.

Ethnopharmacological relevanceDehydrocorydaline (DHC), an active component of Corydalis yanhusuo (Y.H. Chou & Chun C. Hsu) W.T. Wang ex Z.Y. Su & C.Y. Wu (Papaveraceae), exhibits protective and pain-relieving effects on coronary heart disease, but the underlying mechanism still remains unknown. Aim of the studyNetwork pharmacology and experimental validation both in vivo and in vitro were applied to assess whether DHC can treat myocardial ischemia-reperfusion injury (MIRI) by regulating the forkhead box O (FoxO) signalling pathway to inhibit apoptosis. Materials and methodsDHC and MIRI targets were retrieved from various databases. Molecular docking and microscale thermophoresis (MST) determined potential binding affinity. An in vivo mouse model of MIRI was established by ligating the left anterior descending coronary artery. C57BL/6N mice were divided into sham, MIRI, and DHC (intraperitoneal injection of 5 mg/kg DHC) groups. Haematoxylin and eosin, Masson, and immunohistochemical stainings verified DHC treatment effects and the involved signalling pathways. In vitro, H9c2 cells were incubated with DHC and underwent hypoxia/reoxygenation. TUNEL, JC-1, and reactive oxygen species stainings and western blots were used to explore the protective effects of DHC and the underlying mechanisms. ResultsVenny analysis identified 120 common targets from 121 DHC and 23,354 MIRI targets. DHC exhibited high affinity for CCND1, CDK2, and MDM2 (<−7 kcal/mol). In vivo, DHC attenuated decreases in left ventricular ejection fraction and fractional shortening, reduced infarct sizes, and decreased cTnI and lactate dehydrogenase levels. In vitro, DHC alleviated apoptosis and oxidative stress in the hypoxia/reoxygenation model by attenuating ΔΨm disruption; reducing the production of reactive oxygen species; upregulating Bax and CCND1 via the FoxO signalling pathway, as well as cleaved-caspase 8; downregulating the apoptosis-associated proteins Bcl-2, Bid, cleaved-caspase 3, and cleaved-caspase 9; and promoting the phosphorylation of FOXO1A and MDM2. ConclusionBy upregulating the FoxO signaling pathway to inhibit apoptosis, DHC exerts a cardioprotective effect, which could serve as a potential therapeutic option for MIRI.

Bio-accessible milk casein derived tripeptide (LLY) mediates overlapping anti- inflammatory and anti-oxidative effects under cellular (Caco-2) and in vivo milieu

The present study emphasized to explore the toxicity effect of malathion on plants using Allium test. The experiments explored the mitotic inhibition, growth and activity of antioxidant enzymes in roots of Allium cepa at different concentrations (50, 125, 250 and 375 ppm) of malathion under different exposure periods (3, 9 and 18 h). The results revealed that all concentrations of malathion were capable to decline the root growth. Malathion-induced mitotic alterations varying from reduction in mitotic index (MI), relative division rate (RDR) and phase distribution along with large number of chromosomal aberrations. These changes were of varying degree depending on the concentration and treatment period. The roots treated with malathion (375 ppm) showed significant (p ≤ 0.05) higher levels of superoxide dismutase and catalase than the control, while the activity of peroxidase was significantly (p ≤ 0.05) low. At 375 ppm malathion, malondialdehyde content was significantly high (p ≤ 0.05) that was increased with the treatment period. Findings concluded that variations in mitotic index, chromosomal aberrations, alterations in malondialdehyde content and activities of antioxidant enzyme could serve as the useful indicators for monitoring the effects of malathion exposures in the real scenario. Superoxide dismutase and catalase enzyme play vital roles in the antioxidant defence mechanisms under malathion toxicity. According to the data on the malondialdehyde content show malathion to be capable of producing superoxide radicals indirectly, and to result in membrane damage and oxidative stress.

Oxidative stress plays the central role in the pathogenesis and progression of diabetic complications. The present study aims to investigate the beneficial effect of oral administration of flavone baicalein in streptozotocin-nicotinamide (STZ-NA) induced diabetic rats by measuring oxidative stress markers, antioxidant enzyme activities and expression analysis of antioxidant genes. Experimental diabetes was induced by a single intraperitoneal (i.p.) injection of STZ (55 mg /kg b.wt), 15 min after the i.p. administration of NA. At the end of the experimental period, thiobarbituric acid reactive substances (TBARS), activities of antioxidant enzymes and expression levels of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and glutathione peroxidase (GPx) were measured in diabetic rats along with serum biochemical parameters namely total cholesterol (TC), total triglyceride (TG), aspartate transaminase (AST) alanine transaminase (ALT) and glycosylated hemoglobin (HbA1c). Oral administration of baicalein (40 mg/kg b.wt/day) demonstrated a significant ameliorative effect on all studied biochemical and oxidative stress parameters. Biochemical findings were corroborated by qPCR expression analysis which showed significant upregulation of antioxidant genes in diabetic rats. These results suggest that baicalein supplementation may reduce diabetes and its complications by suppressing oxidative stress and enhancing gene expression and antioxidant enzyme activities in diabetic rats.

Osmotic and oxidative stress have been implicated in the pathogenesis of diabetic cataract. Nigerloxin, a fungal metabolite, has been shown to possess aldose reductase inhibitory and free radical scavenging potential, in vitro. In the present study, the beneficial influence of nigerloxin was investigated on diabetes-induced alteration in the eye lens of rats treated with streptozotocin. Groups of diabetic rats were administered nigerloxin orally (100mg·(kg body mass)(-1)·day(-1)) for 30days. The activity of lens polyol pathway enzymes(aldose reductase and sorbitol dehydrogenase), lipid peroxides, and advanced glycation end products (AGEs) were increased in the diabetic animals. Levels of glutathione as well as the activity of antioxidant enzymes(superoxide dismutase, glutathione-S-transferase, and glutathione peroxidase) were decreased in the eye lens of the diabetic animals. The administration of nigerloxin significantly decreased levels of lipid peroxides and AGEs in the lens of the diabetic rats. Increase in the activity of aldose reductase and sorbitol dehydrogenase in the lens was countered by nigerloxin treatment. The activity of glutathione and antioxidant enzyme in the lens was significantly elevated in nigerloxin-treated diabetic rats. Examination of the treated rats' eyes indicated that nigerloxin delayed cataractogenesis in the diabetic rats. The results suggest the beneficial countering of polyol pathway enzymes and potentiation of the antioxidant defense system by nigerloxin in diabetic animals, implicating its potential in ameliorating cataracts in diabetics.

Hypothyroidism is one of the most prevalent thyroid pathologies, which causes oxidative stress by disrupting antioxidant mechanisms. In mammals, the thyroid glands regulate metabolism, development, and growth. Dysfunction of the thyroid gland can result in hypothyroidism, hyperthyroidism, thyroiditis, and thyroid cancer. Whey protein is a widely consumed protein supplement containing abundant sulphur-containing amino acids and bioactive peptides. Here, we analysed the effect of whey protein on oxidative stress in hypothyroidism. In vivo studies were conducted in two phases for 30 and 90 days, respectively. Hypothyroidism was induced in Wistar albino rats by administering 0.05% propylthiouracil (PTU) through drinking water. Five hypothyroid groups and the normal control group were maintained in the first 30 day phase of the study. Among these, one group served as the induced control group, three groups received whey protein at different concentrations (100, 300, and 500 mg per kg body weight), and the last group received L-thyroxine (2 μg per 100 g body weight) as a positive medication. The activities of antioxidant enzymes, such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione-S-transferase, were analysed, and the levels of total antioxidants, glutathione, and malondialdehyde were determined. Ca2+ ATPase and Na+/K+ ATPase activities were detected by estimating the inorganic phosphate content. Histopathological analysis was carried out on the thyroid and liver tissues of all groups. Antioxidant activity was notably increased for higher doses of whey protein compared to that in the diseased control group (p < 0.05). From this initial study, the dose that achieved the desired therapeutic effect was 500 mg kg-1, which was considered for the next 90 day phase of the study. The 90 day phase of the study was conducted with five groups: normal, whey protein-supplemented, hypothyroid, whey protein-supplemented hypothyroid, and levothyroxine-supplemented hypothyroid. All the PTU-treated groups showed degenerative alterations in thyroid histology. Whey protein supplementation causes a considerable decrease in MDA levels with an increase in the major antioxidant enzyme and ATPase activities, with p < 0.05. As a nutritional supplement, whey protein, at a 500 mg kg-1 dose, effectively boosts antioxidant activity without causing any toxicological concerns in long-term use.

The activity of antioxidant enzymes, copper-zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD) and catalase (CAT), as well as that of the mitochondrial FAD-dependent a-glycerophosphate dehydrogenase (a-GPD) in the rat interscapular brown adipose tissue (IBAT) were studied after the treatment with methimazole (MMI) for three weeks or with iopanoic acid (IOP) for five days. Besides, the mitochondrial concentration of uncoupling protein-1 (UCP-1) and the activity of catecholamine degrading enzyme monoamine oxidase (MAO) in the IBAT as well as the activity of the catecholamine synthesizing enzyme, dopamine b-hydroxylase (DBH) in rat serum were examined. Judging by the significantly enhanced level of serum DBH, which is an index of sympathetic activity, and that of IBAT MAO, the increase in MnSOD and CAT activities in the IBAT of hypothyroid (MMI-treated) rats seems to be due to elevated activity of sympathetic nervous system (SNS). However, CuZnSOD activity is not affected by SNS. On the contrary, IOP, which is a potent inhibitor of T4 deiodination into T3 producing "local" hypothyroidism, did not change either SNS activity or activities of IBAT antioxidant enzyme. However, both treatments significantly decreased IBAT UCP-1 content and a-GPD activity suggesting that the optimal T3 concentration in the IBAT is necessary for maintaining basal levels of these key mitochondrial parameters.

This study investigates the effects of exogenous abscisic acid in improving cold resistance in two medicinal Astragalus species, A. membranaceus and A. mongholicus, respectively. ABA was applied via foliar spraying at six concentrations (0, 5, 10, 15, 20, and 30 mg/L), after which plants were subjected to 4°C for 72 h. A series of physiological parameters were measured, including relative electrolyte leakage, MDA content, osmotic regulators (soluble sugars, soluble proteins, proline), antioxidant enzyme activities (superoxide dismutase, peroxidase and catalase), and total chlorophyll content. The results indicate that 15 mg/L ABA most effectively alleviated membrane damage and oxidative stress, while also enhancing osmotic adjustment and antioxidant enzyme activities. Both principal component analysis and membership function evaluation consistently identified 15 mg/L as the optimal ABA concentration, with significantly higher comprehensive scores and average membership values compared to other treatments. Interspecific differences were observed between the two species. A. membranaceus exhibited a stronger response in membrane protection and antioxidant enzyme activation, whereas A. mongholicus relied more on osmotic regulation for cold tolerance. These findings demonstrate that 15 mg/L ABA significantly enhances cold resistance in both species and provides a practical and effective strategy for mitigating cold stress in Astragalus cultivation.

To investigate whether miR-124-3p influences cell apoptosis, inflammatory response, and oxidative stress in rats with acute myocardial infarction (AMI) by mediating the SIRT1/FGF21/CREB/PGC1α pathway. A dual-luciferase reporter gene assay was performed to verify the relationship between miR-124-3p and SIRT1. AMI rats were established via coronary artery ligation after injection with agomiR-124-3p, antagomiR-124-3p, and/or SIRT1 siRNA, and triphenyltetrazolium chloride (TTC), HE, and TUNEL stainings were performed. Bio-Plex rat cytokine assays were performed to determine proinflammatory factor levels. qRT-PCR and Western blotting were used to examine the mRNA and protein expression, respectively. The activity levels of antioxidant enzymes in myocardial tissues were also measured. miR-124-3p was confirmed to target SIRT1 in the H9C2 cells. AMI rats exhibited increased miR-124-3p expression and decreased SIRT1 expression in myocardial tissues. HE staining showed a disorganized cell arrangement and inflammatory cell infiltration in the myocardial tissues of the AMI rats, which was more severe in the rats injected with SIRT1 and agomiR-124-3p but was ameliorated in those treated with antagomiR-124-3p. Moreover, the AMI rats in the antagomiR-124-3p group presented with a reduction in infarct area with an increase in antioxidant enzyme activity, Bcl-2 expression, and activation of the FGF21/CREB/PGC1α pathway, as well as a decrease in cell apoptosis rate, Bax and Caspase-3 expression, and levels of proinflammatory factors, effects that were reversed by si-SIRT1. Inhibiting miR-124-3p expression may activate the FGF21/CREB/PGC1α pathway to reduce cell apoptosis, alleviate the inflammatory response, and attenuate oxidative stress in AMI rats by targeting SIRT1. Graphical abstract.

This study aimed to collaboratively investigate the mechanism of variations in intramuscular fat (IMF) content in Wandong cattle using transcriptomics and metabolomics techniques. Longissimus dorsi (LD) muscle samples were collected from thirteen free-range Wandong cattle in Fengyang County, Anhui Province, China. From this initial cohort, eight animals closely matched in age and body weight were selected. Based on IMF content measured by Soxhlet extraction, these eight cattle were divided into two groups: the high-IMF (HF, n = 4) and low-IMF (LF, n = 4) groups. Subsequent analyses were performed on integrated datasets comprising the transcriptome, metabolome, and fatty acid profile. The results revealed a significant increase in IMF in the HF group compared to the LF group (p < 0.05). Specifically, α-linolenic acid (C18:3n3) and γ-linolenic acid (C18:3n6) were significantly more abundant in the LF group compared to the HF group (p < 0.05), whereas oleic acid (C18:1n9c) and cis-9-palmitoleic acid (C16:1) predominated in the HF group. However, saturated fatty acids (SFAs), such as myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), and Margaric acid (C17:0), did not show significant differences (p > 0.05). A total of 9164 differentially expressed genes (DEGs) were identified via transcriptome analysis, with 2202 genes upregulated and 6962 genes downregulated in the HF group compared to the LF group. The expression profiles exhibited a distinct pattern, characterized by the upregulation of genes such as FABP1, SREBF1, and LIPE, while genes including SCD, PPARGC1A, and LEP were downregulated. GO enrichment analysis demonstrated that the majority of DEGs were predominantly abundant across 25 distinct functional categories distributed across the three primary ontologies. KEGG pathway analysis further identified 341 significantly enriched signaling pathways in the HF group (p < 0.05), predominantly involving metabolic pathways, FoxO, AMPK, and PPAR signaling pathways. Untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics analysis revealed 404 differential accumulated metabolites (DAMs), with 187 in positive ion mode and 217 in negative ion mode (p < 0.05). These DAMs were notably enriched in pathways such as glycerophospholipid metabolism, terpene and steroid biosynthesis, fatty acid degradation, and fatty acid metabolism. Notably, C16:1, C18:1n9c, arachidonic acid (peroxide free) (C20:4n6), oleoyl-L-carnitine, and linoleoyl-carnitine were identified as key players in lipid metabolism. Integrating transcriptomics with metabolomics data unveiled significant associations between DAMs linked to lipid metabolism and DEGs. Specifically, C18:1n9c exhibited a positive correlation with LPIN3, while C16:1 showed negative associations with PPAP2B, PPAP2A, CDS2, HADHA, LPL, HSD17B12, ELOVL5, ACSL1, and ACOX1, and positive correlations with PLA2G15, CDIPT, AGPSBG1, and GPD1. In summary, the variation in IMF content in Wandong cattle is co-regulated by key genes (SREBF1, ACSL1, SCD) via the AMPK, PPAR, and FoxO signaling pathways, coupled with alterations in specific fatty acid metabolites such as C18:1n9c, C16:1, and C20:4n6. These findings provide critical molecular insights for the genetic selection and breeding of Wandong cattle, which are renowned for their superior meat quality.