Nrf2 mRNA Research Articles

The protection afforded by Berberine against chemotherapy-mediated nephropathy in rats involves regulation of the antioxidant axis.

Doxorubicin (DOX) treatment in cancer patients leads to nephrotoxicity. The nephroprotective effect of Berberine (BBR), a herbal ingredient, is well documented as antioxidant and activation of the Nrf2 signalling. This study aimed to investigate if Nrf2 is a major protective mechanism of BBR in DOX animal models. Rats were divided as (n = 6 each): Control, BBR (100 mg/kg, orally), DOX (15 mg/kg, orally), BBR + DOX, and BBR + DOX + brusatol (0.2mg/kg, i.p./twice per week) (an Nrf2 inhibitor). DOX was given as a single dose (day 10), whereas BBR was administered for 3weeks on a daily basis. BBR reduced tubular degeneration and improved renal markers in DOX-treated rats. It also reduced renal nuclear levels of NF-κB p65, total reactive oxygen species (ROS), lipid peroxides, interleukin-6 (IL-6), and tumour necrosis factor-α (TNF-α), as well as mRNA levels of Bax and cleaved caspase-3. However, BBR stimulated glutathione (GSH) and superoxide dismutase (SOD) levels, the transcription of Bcl2, and the mRNA, total cytoplasmic, and nuclear levels of Nrf2 with no effect on the cytoplasmic keap1 levels. All these effects disappeared by brusatol. In conclusion, BBR prevents DOX-induced renal damage by activating Nrf2.

Hesperidin protects rats' liver and kidney from oxidative damage and physiological disruption induced by nickel oxide nanoparticles.

Background: Nickel oxide nanoparticles (NiO-NPs) have recently been utilized in various advanced industrial fields like lithium-ion micro batteries, nanofibers, electrochromic devices, and several biomedical applications. NiO-NPs are classified as extremely toxic substances as they can cause long-term harm to the environment and aquatic life. Moreover, frequent and prolonged exposure can affect human and animal health, causing skin allergies and major toxic consequences, such as hepatorenal toxicity. Hesperidin (HSP) has been proven to possess anti-inflammatory, antioxidant, and free radical scavenging activities. Objective: This study aimed to investigate the underlying protective mechanisms and effects of HSP against NiO-NPs-induced hepatorenal toxicities in rats. Materials and Methods: Forty male Wistar rats were randomly divided into four groups (n = 10 in each). The first group served as a Control group. For 8weeks, the second group was administered NiO-NPs (100mg/kg/day), and the third group was given HSP (100mg/kg/day) via oral gavage for both groups. The fourth group received NiO-NPs and HSP concurrently in the same oral daily doses and duration as the second and third groups. Results: NiO-NPs administration revealed a significant increase in plasma biomarkers of nephrotoxicity (urea, creatinine) and hepatotoxicity (ALT, AST) in NiO-NPs group compared to Control group (p < 0.05). In addition, NiO-NPs administration resulted in a substantial increase in malondialdehyde levels with a significant drop in catalase activity and GSH content in Group II. Also, a significant decreased expression of Nrf-2 and Bcl-2 mRNA levels and upregulation of TNF-α, NF-kβ and BAX in the liver and kidney of NiO-NPs group were also detected. Histologically, the liver and kidney of rats of NiO-NPs group showed significant histopathological disturbances, with a substantial increase in the proliferating cell nuclear antigen (PCNA) positive hepatocytes and renal tubular cells in the NiO-NPs group compared to Control and HSP groups (p < 0.05). In contrast, concomitant administration of HSP with NiO-NPs in group IV showed a significant biochemical, histopathological, and immunohistochemical improvement compared to NiO-NPs group. Conclusion: Co-administration of HSP with NiO-NPs significantly ameliorated most of the NiO-NPs-induced hepatorenal toxicities in male rats.

Expression of nuclear factor-erythroid 2-related factor 2 (Nrf2) in mouse uterus during the peri-implantation period

ABSTRACT Nuclear factor-erythroid 2-related factor- 2 (Nrf2) is a nuclear transcription factor that facilitates transcription of genes for detoxification enzymes and antioxidant proteins. We investigated the distribution and expression of Nrf2 during the peri-implantation period. We detected Nrf2 in uteri of mice during estrus (control) and on days 1, 4, 5, 6 and 8 of pregnancy using immunohistochemistry, quantitative real-time polymerase chain reaction and western blotting. Nrf2 immunostaining was significantly greater on days 1, 5 and 6 of pregnancy compared to controls, and on days 4 and 8 of pregnancy; western blotting results were consistent with immunohistochemical observations. Nrf2 mRNA levels on days 5 and 8 were significantly higher than for control uteri. Increased expression of Nrf2 on days 1, 5 and 6 of pregnancy may be important for uterine receptivity, implantation and decidualization by protecting the developing embryo and uterus from the adverse effects of oxidative stress.

Effects of Age and Lifelong Moderate-Intensity Exercise Training on Rats' Testicular Function.

Aging is associated with testicular morphological and functional alterations, but the underlying molecular mechanisms and the impact of physical exercise are poorly understood. In this study, we examined the effects of age and lifelong moderate-intensity exercise on rat testis. Mature adults (35 weeks) and middle-aged (61 weeks) Wistar Unilever male rats were maintained as sedentary or subjected to a lifelong moderate-intensity treadmill training protocol. Testis weight and histology, mitochondrial biogenesis and function, and proteins involved in protein synthesis and stress response were evaluated. Our results illustrate an age-induced testicular atrophy that was associated with alterations in stress response, and mitochondrial biogenesis and function. Aging was associated with increased testicular levels of heat shock protein beta-1 (HSP27) and antioxidant enzymes. Aging was also associated with decreased mRNA abundance of the nuclear respiratory factor 1 (Nrf1), a key transcription factor for mitochondrial biogenesis, which was accompanied by decreased protein levels of the oxidative phosphorylation system (OXPHOS) complexes subunits in the testes of older animals. On the other hand, exercise did not protect against age-induced testicular atrophy and led to deleterious effects on sperm morphology. Exercise led to an even more pronounced decrease in the Nrf1 mRNA levels in testes of both age groups and was associated with decreased mRNA abundance of other mitochondrial biogenesis markers and decreased protein levels of OXPHOS complexes subunits. Lifelong moderate-intensity exercise training was also associated with an increase in testicular oxidative stress markers and possibly with reduced translation. Together, our results indicate that exercise did not protect against age-induced testicular atrophy and was not associated with beneficial changes in mitochondria and stress response, further activating mechanisms of protein synthesis inhibition.

Evaluation of the Levels of Metalloproteinases as well as Markers of Oxidative Stress and Apoptosis in Lung Tissues After Malathion and Rutin Administrations to Rats

Malathion (MLT) is an important environmental pollutant in the organophosphate class. Rutin (RUT), on the other hand, is one of the flavonoid family members whose effectiveness against various toxic agents has been extensively studied. In the present study, the effects of MLT and RUT treatments on oxidative stress, apoptosis and metalloproteinases in lung tissues of rats were investigated. In the study, MDA, GSH, Nrf2, HO-1, MMP2, MMP9 and caspase-3 levels in lung tissues were analyzed by biochemical or RT-PCR method after rats received MLT and/or RUT treatment for 28 days. The data showed that MLT-induced MDA levels decreased after RUT treatment. In addition, it was determined that Nrf2 and HO-1 mRNA transcript levels and GSH levels suppressed by MLT approached the control group levels after RUT treatment. MLT up-regulated the expression of metalloproteinases (MMP2 and MMP9) in lung tissues, while RUT down-regulated the expression of these genes. In addition, it was observed that MLT triggered caspase-3 expression, while RUT exerted an anti-apoptotic effect by suppressing caspase-3. As a result, it was determined that while MLT showed toxic effects in the lung tissues of rats through oxidative stress, apoptosis and metalloproteinases, RUT could alleviate these toxic effects.

The beneficial effects of traditional Chinese medicine on antioxidative status and inflammatory cytokines expression in the liver of piglets.

Oxidative stress and inflammation seriously affected the growth and development of piglets. Traditional Chinese medicine (TCM) prescriptions has been used to prevent various diseases of piglets, including anti-inflammatory and antioxidant. Here, we identified the effects of Xiao-Jian-Zhong-Tang (XJZT) and Jingsananli-sepsis (JJS) on the oxidative stress and inflammatory in the liver of piglets. The piglets were fed with the basal diet (Control group), basal diet affixed with 10 g/kg XJZT (TCM I group), and basal diet affixed with 3 g/kg JJS (TCM II group), respectively. The serum was gathered on days 30 and 60 and the liver samples were also collected on day 60. Results showed that the TCM I and TCM II markedly increased the activities of the glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC), and reduced the levels of malonaldehyde (MDA), TNF-α, IL-6, and IL-8 in serum. In addition, compared to the control group, Nrf2, SOD-1, NQO-1, and HO-1 mRNA expression levels and the protein levels of Nrf2 and HO-1 were significantly increased while NF-κB, TNF-α, IL-6, and IL-8 mRNA expression levels and the phosphorylation levels of NF-κB and IκB-α were decreased in TCM I and TCM II groups. Collectively, these findings suggested that TCM I and TCM II could enhance anti-oxidative and anti-inflammatory capabilities in the liver of piglets via the Nrf2/NF-κB pathway, providing a basis for the functional exploration of TCM prescriptions.

Oxymatrine Alleviates Gentamicin-Induced Renal Injury in Rats.

Gentamicin is an aminoglycoside antibiotic commonly used to treat Gram-negative bacterial infections that possesses considerable nephrotoxicity. Oxymatrine is a phytochemical with the ability to counter gentamicin toxicity. We investigated the effects and protective mechanism of oxymatrine in rats. The experimental groups were as follows: Control, Oxymatrine only group (100 mg/kg/d), Gentamicin only group (100 mg/kg/d), Gentamicin (100 mg/kg/d) plus Oxymatrine (100 mg/kg/d) group (n = 10). All rats were treated for seven continuous days. The results indicated that oxymatrine alleviated gentamicin-induced kidney injury, and decreased rats’ kidney indices and NAG (N-acetyl-beta-d-glucosaminidase), BUN (blood urea nitrogen) and CRE (creatine) serum levels. The oxymatrine-treated group sustained less histological damage. Oxymatrine also relived gentamicin-induced oxidative and nitrative stress, indicated by the increased SOD (superoxidase dismutase), GSH (glutathione) and CAT (catalase) activities and decreased MDA (malondialdehyde), iNOS (inducible nitric oxide synthase) and NO (nitric oxide) levels. Caspase-9 and -3 activities were also decreased in the oxymatrine-treated group. Oxymatrine exhibited a potent anti-inflammatory effect on gentamicin-induced kidney injury, down-regulated the Bcl-2ax and NF-κB mRNAs, and upregulated Bcl-2, HO-1 and Nrf2 mRNAs in the kidney tissue. Our investigation revealed the renal protective effect of oxymatrine in gentamicin-induced kidney injury for the first time. The effect was achieved through activation of the Nrf2/HO-1 pathways. The study underlines the potential clinical application of oxymatrine as a renal protectant agent for gentamicin therapy.

The measurement of NRF2 and TP53 in blood expects radiotherapeutic sensitivity in patients with esophageal cancer

ObjectiveThis study investigates the relationship between the mRNA expression of nuclear factor erythroid 2-related factor 2 (NRF2) and Tumor protein p53 (TP53) in circulating tumor cells (CTC) and sensitivity to radiotherapy in patients with esophageal cancer. To investigate the relationship between cytokines IL-6, CD8+, and NRF2 during patient treatment and their predictive role for treatment. MethodsRadiosensitivity was assessed by measuring a morphological or functional change in the tumor in response to ionizing radiation. Fasting venous anticoagulated blood (EDTA anticoagulation) was drawn from patients, and the Trizol-chloroform two-step method was used for RNA extraction. Data were collected from 45 patients admitted with radiotherapy alone from January 2018 to December 2021. The expression levels of NRF2mRNA (Messenger Ribose Nucleic Acid) and TP53mRNA in CTCs were detected by reverse transcription-polymerase chain reaction (RT-PCR). Pre- and post-treatment changes in IL-6 and CD8+ were recorded. The correlation between their expression level and the clinical stage, radiotherapy sensitivity, and efficacy of patients was analyzed. ResultsTwenty-six cases were sensitive to radiotherapy, and 19 were resistant, for a radiotherapy sensitivity rate of 58.8%. NRF2mRNA and TP53mRNA values increased in 19 radiotherapy-resistant patients and decreased in 26 radiotherapy-sensitive patients compared with those before radiotherapy (P = 0.001, P＜0.05). The ΔCT values of NRF2mRNA and TP53mRNA before treatment were moderately correlated with prognosis (P < 0.002). Inflammatory cytokine IL-6 was elevated in 22 of 45 patients after radiation, P = 0.04. NRF2 mRNA level was consistently elevated with CD8+ in 10 patients, P = 0.02. ConclusionsThe expression of NRF2mRNA and TP53mRNA in the CTCs found in the peripheral blood of patients with esophageal squamous carcinoma was significantly associated with the sensitivity to radiotherapy. NRF2 mRNA level was consistently elevated with CD8+ and IL-6 in patients.

Curcumin alleviates aristolochic acid nephropathy based on SIRT1/Nrf2/HO-1 signaling pathway

Aristolochic acid I (AA-I), presenting in a variety of natural medicinal plants, which could cause tubular epithelial cell injury. Curcumin (CUR), a polyphenolic substance isolated from turmeric, is a natural antioxidant. The aim of this experiment was to investigate whether CUR attenuated AA-I-induced renal injury in rats through the SIRT1/Nrf2/HO-1 signaling pathway. SD rats were treated with AA-I (10 mg/kg) or/and CUR (200 mg/kg) for 28 days to assess the protective effect of CUR on AA-I-induced renal injury in vivo. NRK-52E cells were treated with AA-I (40 μ M) or/and CUR (20 μ M) for 24 h in vitro. The intervention pathway of CUR against oxidative stress injury induced by AA-I was assessed by observing pathological changes, oxidative stress status, apoptosis and the expression of SIRT1/Nrf2/HO-1 signaling pathway-related factors. The results showed that AA-I exposure increased the contents of BUN, Cr, KIM-1, NGAL, ALT and AST in serum. It increased the content of MDA, decreased the activities of SOD, GST, GSH and the content of ATP in renal tissue. Pathological changes such as inflammatory cell infiltration and mitochondrial injury occurred in renal tissue. AA-I exposure resulted in a substantial rise in the levels of BAX, Ccaspase-9, Cleaved Caspase-9, Caspase-3, Cleaved Caspase-3 and a significant decrease in mRNA and protein expression levels of Bcl-2, SIRT1, Nrf2, NQO1, HO-1 and Keap1. However, these changes were reversed by CUR intervention. In summary, AA-I exposure caused mitochondrial dysfunction and triggered apoptosis through the oxidative stress pathway. However, CUR could reduce AA-I-induced renal injury by activating the SIRT1/Nrf2/HO-1 signaling pathway.

Rhizoma curcumae Longae ameliorates high dietary carbohydrate-induced hepatic oxidative stress, inflammation in golden pompano Trachinotus ovatus

In general, starch, as a complex carbohydrate, is the most economical energy source in aquaculture for its relatively low cost. However, excessive dietary levels of carbohydrate result in pathological conditions. An 8-week feeding trial with CT (control diet, containing 21% carbohydrate), HC (a high-carbohydrate diet, containing 50% carbohydrate) and HCR (a HC diet supplemented with 0.015% Rhizoma curcumae Longae) was performed to investigate the protective effect of curcumin on high-carbohydrate-induced hepatic oxidative stress and intestine lesion in juvenile Trachinotus ovatus. In the current study, HC group significantly decreased WGR, SGR, plasma CAT activity, intestinal C4 levels, hepatic Nrf2, Keap1, Bach1, HO1, CAT, and GPX mRNA expression as well as ZO-1, Occludin, and Claudin-3, TGF-β mRNA transcription levels, while the opposite was true for plasma AST activity, hepatic MDA contents, intestinal Claudin-15, NF-κB, IL-1β, IL-6, and TNF-α mRNA expression. In contrast with the HC group, the HCR group significantly increased the activities of hepatic CAT, SOD, intestinal C3, C4, IgG and LZM levels, hepatic Nrf2, Bach1, CAT, and GPX mRNA expression as well as intestinal ZO-1, Occludin, Claudin-3, TGF-β and IL-10 mRNA expression levels, but the opposite trend was found in plasma triglyceride content, hepatic lipid deposition, hepatic Keap1 mRNA level as well as intestinal NF-κB, IL-6. In conclusion, high-carbohydrate diet can cause detrimental effect on physiological health status in Trachinotus ovatus, while adding Rhizoma curcumae Longae can improve hepatic and intestinal health status via attenuating the oxidative stress, inflammation, and reducing lipid deposition.

Effect of electroacupuncture combined with Zhuang-medicine-thread moxibustion on oxidative stress in gastric antrum of diabetic gastroparesis rats

To observe the effect of electroacupuncture (EA) combined with Zhuang-medicine-thread moxibustion on oxidative stress-related indicators in diabetic gastroparesis (DGP) rats, so as to explore its mechanism underlying improvement of DGP. Male SD rats were randomly divided into normal, model, medication, EA, Zhuang-medicine-thread moxibustion (moxibustion) and EA+Zhuang-medicine-thread moxibustion (combination) groups (15 rats in each group). The DGP model was established by intraperitoneal injection of streptozotocin (STZ). Rats of the medication group were treated by gavage of 0.15 mg/mL mosapride citrate suspension（10 mL/kg）. EA (10 Hz/50 Hz, 2 mA, 20 min) or Zhuang-medicine-thread moxibustion (3 cones) was applied to "Zhongwan" (CV12), bilateral "Neiguan" (PC6) and bilateral "Sanyinjiao" (SP6) of the related groups, once a day for 3 weeks. The body weight, blood glucose, gastric emptying rate and intestinal propulsion rate of rats were measured. The serum malondialdehyde (MDA) content was measured by thiobarbituric acid method, the serum supero-xide dismutase (SOD) activity was measured by xanthine oxidase method, and the serum reactive oxygen species (ROS) activity was detected by ELISA. HE staining was used to observe the pathological changes of gastric antrum. The expression of heme oxygenase-1 (HO-1), nuclear factor erythroid 2-related factor 2 (Nrf2), nicotin-amide adenine dinucleotide phosphate oxidases (NOX4), peroxisome proliferators activated receptor-gamma coactivator 1α(PGC-1α) proteins and mRNAs in gastric antrum was detected by Western blot and real-time quantitative PCR, respectively. Compared with the normal group, the body weight, gastric emptying rate, intestinal propulsion rate, serum SOD activity, the expressions of HO-1, PGC-1α, total Nrf2 proteins and mRNAs, and Nrf2 nuclear translocation in gastric antrum were decreased (P<0.01), while the blood glucose, serum MDA content and ROS activity, NOX4 protein and mRNA expressions in gastric antrum were increased (P<0.01) in the model group. Compared with the model group, the blood glucose was decreased in the EA, moxibustion and combination groups (P<0.01); the body weight, gastric emptying rate, intestinal propulsion rate, and the expressions of HO-1 and PGC-1α mRNAs in gastric antrum were all increased in the four treatment groups (P<0.01, P<0.05), while the serum MDA content and ROS activity, NOX4 protein and mRNA expressions in gastric antrum were all decreased (P<0.01); the serum SOD activity and total Nrf2 protein expression in gastric antrum were increased in medication, moxibustion and combination groups (P<0.01, P<0.05); the expressions of HO-1 and PGC-1α proteins, and Nrf2 nuclear translocation in gastric antrum were increased in medication and combination groups (P<0.05, P<0.01); the expression of Nrf2 mRNA was increased in the medication, EA and combination groups (P<0.01, P<0.05). Compared with the combination group, the body weight, gastric emptying rate and intestine propulsion rate were decreased in the medication, EA and moxibustion groups(P<0.01, P<0.05), and the blood glucose increased (P<0.01); the serum MDA content and ROS activity, NOX4 protein and mRNA expressions in gastric antrum were increased (P<0.01, P<0.05), serum SOD activity, and the expressions of total Nrf2 protein, PGC-1α protein and mRNA, HO-1 mRNA in gastric antrum were decreased (P<0.01, P<0.05) in the EA and moxibustion groups; the expression of Nrf2 mRNA was decreased in the moxibustion group (P<0.05). HE staining showed a large number of inflammatory cell infiltration in the lamina propria and submucosa, and the gastric glands in the lamina propria were significantly expanded, the submucosa was severely edematous in the model group, which were relative milder in the four treatment groups. EA combined with Zhuang-medicine-thread moxibustion can effectively improve the activity of antioxidant enzymes, reduce the production of lipid peroxide, and regulate the expression of antioxidant related proteins and genes, which may be one of the mechanisms in treating DGP.

Preventive Electroacupuncture Alleviates Oxidative Stress and Inflammation via Keap1/Nrf2/HO-1 Pathway in Rats with Cyclophosphamide-Induced Premature Ovarian Insufficiency.

Electroacupuncture (EA) is a popular therapeutic therapy for premature ovarian insufficiency (POI). However, little has been known about the underlying processes of EA therapy. To investigate the benefit of EA and reveal the mechanism, thirty SD female rats were allocated into the control, model, sham, EA, and GnRHa groups at random. Vaginal smears were used to monitor the rats' estrous cycle. Serum liver and renal function (ALT, AST, BUN, and Cr), sex hormone (FSH, E2, and AMH), oxidative stress markers (SOD, GSH, and MDA), and inflammatory cytokine (IL6, IL1β, and TNFα) levels were measured by enzyme-linked immunosorbent assay (ELISA). Their ovary morphology was observed by hematoxylin-eosin staining. Transmission electron microscope was used to remark the ultrastructure of the granulocytes. Protein and gene expressions of Keap1/Nrf2/HO-1 pathway were detected by western blot and RT-PCR. Compared with the model group, in the EA group, the levels of serum sex hormones recovered to normal levels. Moreover, it reduced oxidative stress in rats, as demonstrated by increased SOD and GSH levels and decreased MDA levels. Meanwhile, Keap1 mRNA and protein expression dropped considerably in the EA group, while the mRNA and protein expressions of Nrf2 and HO-1 increased. We found that preventive EA might rescue rats with CTX-induced ovarian dysfunction. The anti-inflammatory and antioxidative stress properties of EA, which elevated the Keap1/Nrf2/HO-1 signaling pathway, might be the underlying mechanism. Furthermore, as compared to GnRHa, electroacupuncture did not raise the burden of the liver (ALT and AST) or the kidney (BUN and Cr). Electroacupuncture has a meaningful impact and a sufficient level of safety, making it useful for therapeutic setting in POI.

Lycopene Effects on Metabolic Syndrome and Kidney Injury in Rats Fed a High-Fat Diet: An Experimental Study.

The frequency of overweight and obesity is rising globally. These disorders are prevalent health problems. It has a substantial correlation with a number of health issues, including cardiovascular, metabolic, and diabetes mellitus disorders. Lycopene (Lyc) is an acyclic structural isomer of β-carotene and has powerful antioxidant properties with various promising therapeutic effects. In this study, rats fed a high-fat diet were examined to determine how lycopene affected metabolic syndrome and kidney damage. After being acclimated, rats were divided into 5 groups (n = 8/group) as follows: the first group served as the control and was fed on a normal pelleted diet (4.25% fat) until the end of the experiment. The second group (high-fat diet; HFD) was fed on a high-fat diet (45.5 kcal% fat) composed of 24% fat, 24% protein, and 41% carbohydrate. The third and fourth groups were fed on HFD and administered lycopene at 25 and 50 mg/kg bodyweight orally every day. The fifth group (standard drug group) received HFD and simvastatin (SVS; 10 mg/kg bodyweight orally daily) for 3 months. Tissue samples from the kidney were taken for determination of the biochemical parameters, lipid peroxidation (LPO), protein carbonyl (PC), reduced glutathione (GSH), total thiol group, antioxidant enzymes, namely, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR), in addition to renal mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2), renal levels of inflammatory markers [tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)], and apoptotic markers (BCL2 Associated X (Bax), B-cell lymphoma 2 (Bcl-2), and Bax/Bcl-2 ratio). When compared to the control group, the HFD group’s food consumption, body weight, serum levels of glucose, uric acid, creatinine, LPO, PC, TNF-α, IL-1β, Bax, and the Bax/Bcl-2 ratio all increased significantly. In the kidney sample of HFD-fed rats, there was a downregulation of Nrf2 mRNA expression along with a significant reduction in the enzymatic activity of SOD, CAT, GR, and GPx. Lyc treatment was able to successfully reverse HFD-mediated changes as compared to the HFD group. Consuming lyc helps to prevent fat and renal damage in a positive way.

IGF2BP3-NRF2 axis regulates ferroptosis in hepatocellular carcinoma

Poor sensitivity to sorafenib has been an important constraint on the efficacy of targeted therapy in advanced hepatocellular carcinoma (HCC). Therefore, it is particularly important to explore effective therapeutic targets to improve the sensitivity of HCC cells to sorafenib. Upregulation of IGF2BP3 is strongly associated with tumor invasion, early recurrence and poor prognosis in various human cancers, including HCC, but its roles in the sorafenib treatment of HCC remain unclear. In our study, IGF2BP3 knock-down significantly promoted ferroptosis in HCC cells through the evaluation of the Reactive Oxygen Species (ROS), Fe2+ and malondialdehyde (MDA) levels after sorafenib administration. In addition, NRF2 mRNA was identified as an important target of IGF2BP3 by bioinformatics analysis, RNA binding protein immunoprecipitation (RIP) and RNA pulldown experiments. More importantly, IGF2BP3, as an m6A (N6-Methyladenosine) reader, was shown to promote the stability of NRF2 mRNA by reading its m6A modification. Similar results were obtained from in vivo experiments. In summary, our study uncovered the role of IGF2BP3-NRF2 axis on ferroptosis in HCC, providing significant evidence for new anti-cancer strategies aimed at improving the efficacy of sorafenib.

Dietary effects of aucubin on growth and flesh quality of grass carp (Ctenopharyngodon idellus) based on metabolomics.

This aim of this study was to investigate the effects of dietary aucubin on the growth, flesh quality, and metabolomics of grass carp (Ctenopharyngodon idella). Five diets were designed with the aucubin inclusion of 0 (control diet), 0.2, 0.4, 0.6, and 0.8 g/kg (Auc-0.2, Auc-0.4, Auc-0.6, Auc-0.8) and were fed to grass carp with an initial body weight of 17.0 ± 0.2 g for 60 d. The results indicated that dietary aucubin did not significantly affect the growth performance of grass carp (P > 0.05). Compared to the control, dietary supplementation with 0.2 to 0.8 g/kg aucubin increased flesh hardness, chewiness, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and the contents of total free amino acids (TFAA) and n-3 polyunsaturated fatty acids (n-3 PUFA) (P < 0.05). The contents of malondialdehyde (MDA) and lactic acid (LD) in the flesh were significantly decreased by the addition of 0.4 to 0.6 g/kg aucubin and by the addition of 0.6 to 0.8 g/kg aucubin (P < 0.05), respectively, while the content of delicious amino acids (DAA) was significantly enhanced by the addition of 0.4 to 0.8 g/kg aucubin (P < 0.05). Moreover, the contents of collagen and C22:6n3 (DHA) in the flesh of the Auc-0.8 group were significantly higher than those of the control (P < 0.05). In the metabolomics profiling of flesh, 133 and 135 named differential metabolites were identified in the Auc-0.4 and Auc-0.8 groups, respectively, compared to the control, and these metabolites were found to be involved in the second-grade pathways of "lipid metabolism" and "amino acid metabolism". Regarding gene expression, the mRNA levels of CuZn-SOD, CAT, COL1A1, COL1A2, Smad4, and FAS in flesh were upregulated in the Auc-0.4 and Auc-0.8 groups, and the expression levels of GPx, Nrf2, and TGF-β1 mRNA were also upregulated in the Auc-0.8 group (P < 0.05). In summary, dietary aucubin did not promote growth, but improved the flesh quality of grass carp, which might be associated with the TGF-β/Smad and Nrf2 pathways. The recommended supplementation level of aucubin in the diet of grass carp was 0.6 to 0.8 g/kg.

Effect and mechanisms of Polygonatum kingianum (polygonati rhizome) on wound healing in diabetic rats

Ethnopharmacological relevanceDiabetic dermatopathy is one of the most serious and common complications of diabetes. It has been found that high glucose can lead to abnormal glycometabolism. The skin microenvironment pollution caused by the increase in glucose and the oxidative stress mediated by the deposition of advanced glycation end products can lead to invisible skin injury, and the interaction between them is the key factor that makes the skin wounds of diabetic rats difficult to heal. Therefore, the main task of promoting healing is to reduce blood glucose levels and relieve the deposition of advanced glycation end products. Polygonatum kingianum Collett & Hemsl (PK) of Asparagaceae is planted in Yunnan, China, and is used by the Bai, Hani and Wa nationalities as a traditional medicine for preventing and treating diabetes. Aim of the studyTo study the effects of PK extract on skin wound healing in diabetic rats and to explore the regulatory mechanism of PK on wound microenvironment pollution, the antioxidative stress signaling pathway and latent injury of wound skin tissue. MethodsFirst, wounds were prepared after diabetic rats were given PK extract by gavage for 4 weeks, and then gavage was continued for 2 weeks to observe and calculate the wound healing rate. A scanning electron microscope was used to observe the pathomorphological changes in the skin tissue at the edge of the wound. Western blotting was used to detect protein expression. Immunohistochemistry was used to detect the expression of CD34, AGEs, bFGF and VEGF. The Nrf2/HO-1 signaling pathway in skin tissue was detected by fluorescence quantitative PCR. Serum biochemical indicators and inflammatory cytokine levels were detected by a kit. ResultsAfter PK treatment, the wound healing rate increased significantly (P < 0.001), the infiltration of inflammatory cells in skin tissue of DM lesion rats decreased, the number of new blood vessels increased, and the epidermis and dermis thickened. The content of glucose, AGEs, RAGE protein and RAGE mRNA in skin decreased significantly (P < 0.05, P < 0.01, P < 0.001), while the expression of Nrf2 mRNA, HO-1 mRNA, CD34, bFGF and VEGF increased significantly (P < 0.05, P < 0.01, P < 0.001). The levels of SOD, GSH, MMP-9 and MMP-2 in skin decreased (P < 0.05, P < 0.01, P < 0.001), but the level of TIMP-2 increased (P < 0.001). GSP, GHb and ICAM-1 in plasma decreased (P < 0.05, P < 0.01, P < 0.001), while T-AOC, SOD and FINS increased (P < 0.05, P < 0.01). The levels of MDA, TNF-, IL-6, IL-2 and IFN-γ in plasma and wound skin tissue decreased (P < 0.05, P < 0.01, P < 0.001). ConclusionPK can reduce the infiltration of inflammatory cells and glucose content in the skin tissue at the edge of the wound, reduce inflammatory factors in skin and plasma, and increase angiogenesis, thus improving the wound healing rate. PK can alleviate the microenvironment pollution caused by AGEs and glucose metabolism disorder in diabetic rats and induce antioxidant activity through the Nrf 2/HO-1 signaling pathway, thus reducing oxidative damage and offsetting endogenous skin damage and hidden damage.

Alleviating effect of quercetin on cadmium-induced oxidative damage and apoptosis by activating the Nrf2-keap1 pathway in BRL-3A cells.

Cadmium (Cd) is a toxic heavy metal extensively used in industrial and agricultural production. Among the main mechanisms of Cd-induced liver damage is oxidative stress. Quercetin (QE) is a natural antioxidant. Herein, the protective effect of QE on Cd-induced hepatocyte injury was investigated. BRL-3A cells were treated with 12.5 μmol/L CdCl2 and/or 5 μmol/L QE for 24 h. The cells and medium supernatant were collected, and the ALT, AST, and LDH contents of the medium supernatant were detected. The activities or contents of SOD, CAT, GSH, and MDA in cells were determined. Intracellular ROS levels were examined by flow cytometry. Apoptosis rate and mitochondrial-membrane potential (ΔΨm) were detected by Hoechst 33,258 and JC-1 methods, respectively. The mRNA and protein expression levels of Nrf2, NQO1, Keap1, CytC, caspase-9, caspase-3, Bax, and Bcl-2 were determined by real-time PCR (RT-PCR) and Western blot methods. Results showed that Cd exposure injured BRL-3A cells, the activity of antioxidant enzymes decreased and the cell ROS level increased, whereas the ΔΨm decreased, and the expression of apoptotic genes increased. Cd inhibited the Nrf2-Keap1 pathway, decreased Nrf2 and NQO1, or increased Keap1 mRNA and protein expression. Through the combined action of Cd and QE, QE activated the Nrf2-Keap1 pathway. Consequently, antioxidant-enzyme activity decreased, cellular ROS level decreased, ΔΨm increased, Cd-induced BRL-3A cell damage was alleviated, and cell apoptosis was inhibited. After the combined action of QE and Cd, Nrf2 and NQO1 mRNA and protein expression increased, Keap1 mRNA and protein expression decreased. Therefore, QE exerted an antioxidant effect by activating the Nrf2-Keap1 pathway in BRL-3A cells.

Arginine Enhances Ovarian Antioxidant Capability via Nrf2/Keap1 Pathway during the Luteal Phase in Ewes.

Simple SummaryThe quantity and quality of forage grass may decrease to some extent in autumn and winter, and feeding with such forage grass may temporarily cause malnutrition and ovarian oxidative stress in ewes. Arginine is one of the most abundant amino acids enhancing the antioxidant capability in animal tissues. Thus, this study investigated the effects of arginine on ovarian antioxidant capability during the luteal phase in ewes. Our results showed that nutrition restriction can reduce the ovarian antioxidant capability in ewes, while arginine supplementation can significantly improve it, which was associated with the nuclear factor erythroid 2-related factor 2/Kelch-like ECH-associated protein 1 pathway.This study evaluated the effect of arginine (Arg) on ovarian antioxidant capability during the luteal phase in ewes. A total of 108 multiparous Hu sheep at two years of age were randomly allocated to three groups: a control group (CG), a restriction group (RG), and an Arg group (AG), with six replicates per group and six ewes per replicate. Our results showed that the end body weight was significantly decreased in the RG group (p < 0.05), while the Arg addition reversed this reduction. The estrous cycle days were significantly increased in the RG group (p < 0.05), while Arg addition reversed this time extension. Compared with the control group, restricting feeding could significantly enhance the number of small follicles (SF), total follicles (TF), large corpora lutea, and the SF/TF (p < 0.05), while Arg addition reduced the number of SF and TF. However, the large follicles/TF were significantly decreased (p < 0.05), while Arg addition reversed this reduction. In addition, nutrition restriction significantly increased the malondialdehyde (MDA) level (p < 0.05), while significantly decreased the glutathione/glutathione disulfide and the activities of superoxidative dismutase, catalase, and glutathione peroxidase in the ovaries (p < 0.05). However, Arg addition reversed this enhancement of the MDA level and the reductions in these antioxidant enzymes activities. In addition, positive relationships occurred between antioxidant enzyme activities and the enzyme mRNA expressions. Meanwhile, the nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA expression was positively connected with antioxidant mRNA expressions and negatively related to the Kelch-like ECH-associated protein 1 (Keap1) mRNA expression. The Nrf2 protein expression was negatively related to the Keap1 protein expression. In conclusion, nutrition restriction reduced the ovarian antioxidant capability in ewes, while this was significantly improved by Arg supplementation, which was associated with the Nrf2/Keap1 pathway.

Influence of zinc levels and Nrf2 expression in the clinical and pathological changes in patients with diabetic nephropathy

ObjectiveWe investigated the correlation between zinc levels and Nrf2 expression and potential effects on the clinicopathology of patients with diabetic nephropathy (DN).MethodsWe selected 30 patients with DN, diagnosed via renal biopsy at our hospital from March 2018 to November 2019, and enrolled 30 healthy individuals from a medical examination center as the control group. Patients with DN were divided into normal-zinc and low-zinc groups. We detected the levels of zinc, copper, and Nrf2 mRNA in their serum, and collected the clinical and pathological data of DN patients.ResultsSerum zinc level and Nrf2 mRNA expression were significantly decreased in patients with DN compared to those of healthy people (P < 0.05). Of the 30 patients, 16 had low zinc (53.3%) and 14 had normal zinc levels (46.7%). There was no significant difference in the blood Nrf2 mRNA expression between the two groups (P > 0.05). However, the expression of Nrf2 in the kidney tissue of the low-zinc group was significantly lower compared to the normal-zinc group (P < 0.05). Diastolic blood pressure and copper levels were significantly higher in the low-zinc group (P < 0.05). In contrast, body mass index, red blood cell count, Hb level, and the ratio of zinc to copper were significantly lower in the low-zinc group (P < 0.05). The pathological classifications of the low-zinc group were more severe (P < 0.05).ConclusionPatients with DN were more likely to have zinc deficiency and lower expression of Nrf2. Additionally, DN patients with zinc deficiency were prone to have more severe clinical and pathological manifestations.

Alleviative effect of biofloc technology (BFT) on extruded soybean meal (ESBM)-induced growth inhibition and intestinal barrier dysfunction in Rhynchocypris lagowskii

This study evaluated the alleviative effect of bioflocs technology (BFT) on intestinal morphology, permeability, digestive function, immune function, antioxidant capacity and microbiota of Rhynchocypris lagowskii sensitized by extruded soybean meal (ESBM). In a 2 × 2 factorial design, R. lagowskii (6.40 ± 0.01 g) was randomly divided into 4 groups, which contains two culture modes (traditional mode and biofloc mode) and two levels of dietary ESBM (0% and 68%). At the end of the experiment (8 weeks), the results showed that the growth performance of R. lagowskii was significantly increased by BFT, but significantly decreased by feeding diets with 68% ESBM (P < 0.05). Culture mode significantly affected intestinal growth indexes, morphology, the levels of tight junction protein mRNA, the activities of digestive enzymes, immune function, inflammation response and antioxidant capacity in proximal intestines (PI), mid intestines (MI) and distal intestines (DI) (P < 0.05). These indexes (except fold height, lamina propria width, the levels of tight junction protein mRNA, the activities of protease, AKP and LZM, the levels of inflammation related mRNA, the levels of T-SOD and PC, the levels of Nrf2, Keap1, HO-1 and Maf mRNA in PI, the activities of CAT in MI, the activities of lipase and amylase in intestines) were also significantly affected the levels of dietary ESBM (P < 0.05). Intestinal permeability (DAO, 5-HT, ET-1 and D-LA) were only significantly affected by the levels of dietary ESBM (P < 0.05). In addition, the interaction between culture mode and the levels of dietary ESBM was observed for muscular layer thickness and T-AOC in PI, fold height, ZO-1, NF-κB and Nrf2 in MI, Claudin-2, IL-1β, TGF-β, T-AOC, Nrf2, HO-1 and Maf in DI (P < 0.05). Results of intestinal microbiota showed that relative abundance of Amaricoccus, Aquitalea, Chelatococcus and Nakamurella were significantly affected by culture mode, the levels of ESBM in diets and their interaction (P < 0.05). Therefore, this study indicated that intestinal morphology, permeability, digestive function, immune function, antioxidant capacity and microbiota in R. lagowskii could be affected by ESBM and alleviated by BFT.