Upregulation Of Expression Research Articles

Inflammation induced PFKFB3-mediated glycolysis promoting myometrium contraction through the PI3K-Akt-mTOR pathway in preterm birth mice.

Inflammation is a significant risk factor for preterm birth. Inflammation enhances glycolytic processes in various cell types and contributes to the development of myometrial contractions. However, the potential of inflammation to activate glycolysis in pregnant murine uterine smooth muscle cells (mUSMCs) and its role in promoting inflammatory preterm birth remain unexplored. In this study, lipopolysaccharide was employed to establish both cell and animal inflammation models. We found that inflammation of mUSMCs during late pregnancy could initiate glycolysis and promoted cell contraction. Subsequently, the inhibition of glycolysis using the glycolysis inhibitor 2-deoxyglucose can reverse inflammation-induced cell contraction. The expression of 6-phosphofructokinase 2 kinase (PFKFB3) was significantly upregulated in mUSMCs following lipopolysaccharide stimulation. Additionally, lactate accumulation and enhanced contraction were observed. Inhibition of PFKFB3 reversed the lactate accumulation and enhanced contraction induced by inflammation. We also found that inflammation activated the phosphatidylinositol 3-kinase (PI3K) - protein kinase B (Akt) - mammalian target of rapamycin (mTOR) pathway, leading to the upregulation of PFKFB3 expression. The PI3K-Akt pathway inhibitor LY294002 and the mTOR pathway inhibitor Rapamycin effectively inhibited the upregulation of PFKFB3 protein expression, lactate production, and the enhancement of cell contraction induced by lipopolysaccharide. This study indicates that inflammation regulates PFKFB3 through the PI3K-Akt-mTOR pathway, which enhances the glycolytic process in pregnant mUSMCs, ultimately leading to myometrial contraction.

WNT3A promotes the cementogenic differentiation of dental pulp stem cells through the FOXO1 signaling pathway

BackgroundDental pulp stem cells (DPSCs) possess capability of multidirectional differentiation, and their cementogenic differentiation potential enables them to participate in cementum repair and regeneration. The molecular mechanisms underlying cementogenic differentiation of DPSCs remain unclear.MethodsDPSC data set GSE138179 was retrieved from gene expression omnibus (GEO) database. Weighted gene co-expression network analysis (WGCNA) was employed to identify significant modules. Pathway enrichment exploration was conducted utilizing gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Set Enrichment Analysis (GSEA), and Metascape tools. CIBERSORT was utilized to analyze immune cell infiltration analysis. The comparative toxicogenomics database (CTD) was utilized for the validation of core targets. Subsequently, cell experiments were conducted to validate the core targets. Changes in protein expression related to the FOXO1 signaling pathway, cell cycle, and apoptosis were evaluated using western blotting (WB).ResultsDifferentially expressed genes (DEGs) associated with DPSC cementogenic differentiation were predominantly enriched in crucial pathways such as the signaling pathway, cell apoptosis, and Wnt signaling pathway. Bioinformatics analysis confirmed WNT3A as a pivotal biomarker for DPSC cementogenic differentiation, and WNT3A was highly expressed in the cementogenic differentiation group. Western blotting results demonstrated that compared to the DPSC group, molecules such as Caspase-3, Caspase-9, FAS, P53, and BAX were downregulated in the CDDPSC group, suggesting reduced apoptosis. Furthermore, upregulation of WNT3A expression in CDDPSC–OE further suppressed the expression of these apoptotic molecules, suggesting a mitigated apoptotic response. Downregulation of WNT3A expression in CDDPSC–KO resulted in increased expression of apoptosis-related molecules, thereby enhancing apoptosis.ConclusionsWNT3A is highly expressed in the cementogenic differentiation of DPSC, and WNT3A mediates FOXO1 pathway to promote differentiation of dental pulp stem cells into cementogenic differentiation, thus realizing the formation and maintenance of cementum tissue.

HMGB1 Promotes Lysosome-Dependent Cell Death Induced Via Dry Eye by Disrupting Lysosomal Homeostasis.

Hypertonic stress can induce apoptosis, inflammation, and dry eye disease (DED) through the upregulation of HMGB1 expression. However, its role in mediating and maintaining lysosomal homeostasis and suppressing DED development in living and in vitro models is unclear. Immunofluorescence, nucleoplasmic separation, and electron microscopic analysis were used to compare the effects of hypertonic stress on lysosomal function, nuclear HGMB1 expression, and lysosomal localization in three different dry eye models. The live model was established by the subcutaneous injection of scopolamine (SCOP) into C57BL/6J female mice, and the in vitro model used human corneal epithelial cell (HCEC) cultures that were maintained at a hyperosmotic level of 450 milliosmolar (mOsm). RNAi technology was used to knockdown HMGB1 expression levels, altering hyperosmotic stress-induced changes in survival and autophagy of corneal epithelial cells in vitro. Lysosomal protease inhibitors were applied to suppress the increase of corneal tissue inflammation in the dry eye mouse model. This hypertrophic stress upregulated karyoplasmic HMGB1 expression in HCECs. This change disrupted lysosomal structural and functional integrity, which facilitated the release of HMGB1 into the cytoplasm. SiRNAs downregulated HMGB1 expression levels and reversed increases in HMGB1 transfer from the nucleus to the cytoplasm and lysosomal alkalinizing function. These changes promoted increases in cathepsin leakage from lysosomes and increased mouse corneal epithelial apoptosis, whereas autophagy decreased. In a mouse model, administration of a cathepsin inhibitor reduced both ocular inflammation and other aspects of dry eye symptomology. Hyperosmotic-induced HMGB1 upregulation and increased HMGB1 transfer into the cytoplasm from the nucleus underlie the loss of lysosomal membrane integrity, which increases both cathepsin release and corneal epithelial apoptosis. HMGB1 downregulation instead ameliorated these pathogenic responses.

Protective Role of Nano-encapsulated Bifidobacterium breve, Bacilllus coagulans, and Lactobacillus plantarum in Colitis Model: Insights Toward Propagation of Short-Chain Fatty Acids and Reduction of Exaggerated Inflammatory and Oxidative Response.

Irritable bowel disease (IBD), also known as ulcerative colitis and Crohn's disease, is a chronic inflammatory disorder affecting millions of people worldwide. Herein, nano-encapsulated multi-strain probiotics formulation, comprising Bifidobacterium breve DSM24732 and B. coagulans SANK 70258 and L. plantarum DSM24730 (BBLNPs) is used as an effective intervention technique for attenuating IBD through gut microenvironment regulation. The efficacy of the prophylactic role of BBLNPs in alleviating injury induced by dextran sulfate sodium (DSS) was evaluated by assessing oxidative and inflammatory responses, levels of short-chain fatty acids (SCFAs) and their regulation on GPR41/43 pathway, expression of genes related to tight-junctions and autophagy, immunohistochemistry of IL1β and GPR43, and histological examination of inflamed colonic tissue. The severity of clinical signs and paracellular permeability to FITC (fluorescein isothiocyanate)-labeled dextran was significantly decreased after BBLNP treatment. Reduction of oxidative stress-associated biomarkers (MDA, ROS, and H2O2) and acceleration of antioxidant enzyme activities (SOD, CAT, and GSH-Px) were noted in the BBLNP-treated group. Subsiding of inflammatory markers (TNF-α, IL-18, IL-6, TRL-4, CD-8, NLRP3, and caspase 1) and upregulation of tight-junction-related genes (occludin and JAM) was detected in BBLNPs. Administration of BBLNPs remarkably resulted in a higher level of SCFAs which parrel with colonic upregulation of GPR41 and GPR43 expression compared to DSS-treated rats. Notable modulation of autophagy-related genes (p62, mTOR, LC3, and Beclin-1) was identified post BBLNP treatment. The mRNA expressions of p62 and mTOR were significantly downregulated, while LC3 and Beclin-1 were upregulated after prophylactic treatment with BBLNPs. Immune-stained labeled cells showed lower expression of IL-1β and higher expression levels of GPR43 in BBLNPs compared to the DSS-induced group. The intestinal damage caused by DSSwas effectively mitigated by oral BBLNP treatment, as supported by the restoration of healthy colonic tissue architecture. The findings suggest that BBLNPs have a promising avenue in the remission of IBD by modulating inflammation, oxidative stress, microbial metabolites such as SCFAs, and autophagy.

KLF12 Aggravates Angiotensin II-Induced Cardiac Remodeling in Male Mice by Transcriptionally Inhibiting SMAD7.

Adverse left ventricular remodeling and subsequent heart failure remain a major cause of patient morbidity and mortality worldwide. The KLF family of transcription factors plays crucial roles in heart injury. KLF12 (Krüppel-like factor 12) is a transcription factor that regulates multiple disease processes, although the specific role of KLF12 in cardiac remodeling remains unclear. In our study, we observed a significant upregulation of KLF12 expression in remodeling hearts. The increased expression of KLF12 primarily originated from cardiac fibroblasts during the fibrotic response induced by angiotensin II. To investigate the effects of KLF12, we performed RNA-seq and found that KLF12 overexpression significantly upregulated the cardiac remodeling associated pathway. Hence, we generated adult mice with cardiac fibroblast-specific overexpression of KLF12 using lentivirus or miRNA (miR-1/133TS) technology. Compared with control mice, KLF12-miR1/133TS transfected mice exhibited exacerbated cardiac remodeling and function. Mechanistically, we discovered that KLF12 directly binds to the promoter of Smad7, leading to the activation of the TGF-β (transforming growth factor beta)-Smad3 pathway. In conclusion, KLF12 promoted the development of angiotensin II-induced cardiac remodeling in male mice. Targeting KLF12 may be a promising therapeutic approach to treat cardiac remodeling.

Smooth muscle cell Piezo1 is essential for phenotypic switch and neointimal hyperplasia.

Disturbed blood flow is a critical factor in activation of vascular smooth muscle cells (VSMCs) and initiation of neointimal hyperplasia. The Piezo1 channel is a recent yet well-characterised mechanosensor that plays a vital role in vascular development and homeostasis. However, the role of VSMC Piezo1 in neointima development remains largely unknown. The purpose of this study is to investigate the functional role of Piezo1 channel in neointimal hyperplasia. We measured the expression of Piezo1 in VSMC-rich neointima from human coronary artery samples and two mouse neointimal hyperplasia models which were induced by cast implantation or guidewire injury. We utilised VSMC-specific Piezo1 knockout mice to explore its function and the underlying mechanism of neointimal hyperplasia, both in vivo and in vitro. In human and mouse neointima samples, we observed a significant up-regulation of Piezo1 expression in the VSMC-rich neointima compared to the medial layer. VSMC-specific knockout of Piezo1 significantly reduced neointimal hyperplasia in both animal models. Activation of Piezo1 facilitates, whereas Piezo1 deficiency inhibits disturbed flow-induced cell proliferation, migration and synthetic phenotype switch. Mechanistic studies suggest that Piezo1 activates YAP and TAZ through Ca2+ and its downstream effectors calmodulin kinase II and calcineurin, which in turn drive VSMC proliferation and migration, thereby facilitating neointimal hyperplasia. These findings demonstrate a critical role of mechanosensitive Piezo1 channel in neointimal hyperplasia via modulating VSMC phenotype. Piezo1 channels may represent a novel therapeutic target for maladaptive vascular remodelling and occlusive vascular diseases.

Evaluation of mucosal adjuvants to chitosan-nanoparticle-based oral subunit vaccine for controlling salmonellosis in broilers

Salmonellosis, a gastrointestinal disease, continues to be one of the major public health concerns worldwide. Poultry meat and eggs are recognized as the major source of Salmonella food poisoning in humans. Our study evaluated the protective efficacy of mannose-conjugated chitosan-nanoparticle (mChitosan-NP)-based subunit vaccine, consisting of immunogenic outer membrane proteins and flagella of Salmonella Enteritidis [mChitosan (OMP+FLA)/FLA-NP], coadministered orally with potent mucosal adjuvants to reduce the colonization of S. Enteritidis in the intestines of broiler chickens. We evaluated the adjuvant effects of three different doses of two well-known mucosal adjuvants, c-di-GMP (stimulator of interferon gene agonist) and whole cell lysate (WCL) of Mycobacterium smegmatis, to improve the efficacy of mChitosan (OMP+FLA)/FLA-NP vaccine. Our data reaffirmed the potent adjuvanticity of both of these adjuvants and identified their optimal dose when entrapped in mChitosan-NP to potentiate the immunogenicity and efficacy of orally delivered mChitosan (OMP+FLA)/FLA-NP vaccine. The physical characteristics of mChitosan (OMP+FLA)/FLA-NP, mChitosan-GMP/FLA-NP, and mChitosan-WCL/FLA-NP formulations revealed a high positive charge (Zeta potential +20–25 mV), size 235–260 nm, and polydispersity index 0.35–0.52, which are conducive for oral delivery. The efficacy in chickens that received oral administration with a combination of the vaccine-adjuvant formulations was evaluated by challenging with Salmonella Enteritidis. Our data showed that mChitosan (OMP+FLA)/FLA-NP WCL at 10 µg/dose formulation consistently reduced the S. Enteritidis load by over 0.5 log10 comparable to a commercial live vaccine at post-challenge days 4 and 10. Immunologically, we observed enhanced systemic and mucosal antibody and cellular (B cells and T-helper cells) immune responses as well as upregulation of expression of immune cytokine genes IFN-γ, TGF-β, and IL-17 in the cecal tonsils of adjuvanted mChitosan-NP Salmonella-subunit-vaccinated birds. Overall, particularly the mucosal adjuvant WCL consistently enhanced the efficacy of mChitosan (OMP+FLA)/FLA-NP vaccine by inducing effective immune responses.

Mode of Action of Retinoic Acid in the Regulation of Glucose Metabolism in Mud Crab, Scylla serrata: Evidence for the Involvement of Crustacean Hyperglycemic Hormone

In the current study, we evaluated the effect of retinoids, 9-Cis retinoic acid (9CRA) and all-trans retinoic acid (ATRA) on carbohydrate metabolism in the mud crab, Scylla serrata. Intact and eyestalk ablated male crabs (n=10 crabs per group) were selected and retinoic acid isomers, 9CRA and ATRA were injected into the crabs. Parallel controls were maintained. Significant reduction in the hemolymph sugar levels was observed in crabs subjected to eyestalk ablation (ESX) over intact crabs. Injection of 9CRA into intact but not in ESX crabs exhibited a dose- and time-dependent manner hyperglycemic response. ATRA administration did not induce hyperglycemia either in intact or ESX crabs. Injection of 9CRA showed a significant decrease in total carbohydrate content and glycogen levels in hepatopancreas and muscle tissues of intact crabs over their respective controls. Further, the activity levels of glycogen phosphorylase were also elevated in selected tissues of intact crabs administered with 9CRA. In addition, intact crabs which received 9CRA resulted in up regulation of expression of CHH mRNA from the eyestalks. Taken together, we postulate that 9CRA-induced hyperglycemia might be ascribed to neurotransmitter effects mediating the release of CHH from the eyestalk. The release of CHH eventually caused glycogenolysis in the selected hepatopancreas and muscle tissues of the mud crab Scylla serrata thereby hyperglycemia.

Ganoderic acid a potential protective impact on bleomycin (BLM) -induced lung fibrosis in albino mice: Targeting caveolin 1/TGF-β/ Smad and P38MAPK signaling pathway.

Bleomycin (BLM), an anticancer medication, can exacerbate pulmonary fibrosis by inducing oxidative stress and inflammation. Anti-inflammatory, anti-fibrotic, and antioxidant properties are exhibited by ganoderic acid A (GAA). So, we aim to assess GAA's protective impact on lung fibrosis induced via BLM. Forty mice were randomly classified into four groups. Lung fibrosis was induced by injection of BLM intraperitoneally (15mg/kg body weight). GAA was given by oral gavage (25mg/kg body weight). Lung tissue MDA, TAC, and GSH were assessed spectrophotometrically. As well, TGFβ, p38 MAPK, TNF-α, IL-1β, and CAV1 levels were measured by enzyme-linked immunosorbent assay. Gene expression of tumor growth factor beta (TGF-β), Smad2, Smad3, and glutamate-cysteine ligase (GCL) were also evaluated. GAA had significantly improved biochemical biomarkers as well as histopathology of the lung. The protective impact of GAA may be linked to the upregulation of GCL gene expression and subsequent GSH levels. In addition, the GAA-treated group showed a significant decrement in the levels of TGF-β, Smad2&3, P38 MAPK, TNF-α, IL1β, and MDA compared to BLM induced lung fibrosis group. GAA has a protective impact on lung fibrosis induced by BLM via downregulation of TGF-β and upregulation of CAV1 level and GCL expression which may play a critical role in the improvement of the pathogenesis of lung fibrosis induced via BLM.

4-amino-3-(phenylselanyl) benzenesulfonamide attenuates intermittent cold stress-induced fibromyalgia in mice: Targeting to the Nrf2-NFκB axis

Stress is widely recognized as the primary environmental factor associated with chronic pain conditions, including fibromyalgia. A recent study demonstrated the potential antinociceptive effects of 4-amino-3-(phenylselanyl) benzenesulfonamide (4-APSB) in acute nociceptive animal models due to its antioxidant and anti-inflammatory properties. However, the efficacy of 4-APSB in managing chronic painful conditions, such as fibromyalgia, has not been explored so far. This study investigated the pharmacological effects of 4-APSB in an experimental model of fibromyalgia induced by intermittent cold stress (ICS). Male and female mice were divided into Control, ICS, 4-APSB, and ICS + 4-APSB. After the ICS, the animals were treated with 4-APSB (1 mg kg−1) or vehicle by the intragastric route until the tenth day. The behavioral tasks were performed on days 5, 8, and 10. The findings showed a negative correlation between paw withdrawal threshold and Nrf2 or NFκB mRNA expression levels caused by ICS exposure. The 4-APSB suppressed the nociceptive signs and a depressive like-phenotype in male and female mice exposed to ICS. 4-APBS normalized the elevated levels of TBARS and the up-regulation of Nrf2 and NFκB expression in the cerebral cortex of ICS-exposed mice. This compound also modulated the oxidative stress in the spinal cord of female mice. The 4-APSB attenuated the inhibition of Na+, K+ − ATPase activity in the central nervous system (CNS) of female mice exposed to ICS. 4-APSB attenuated behavioral and redox imbalance triggered by the ICS model in male and female mice, suggesting its beneficial effects for treating fibromyalgia in both sexes.

Hesperidoside abolishes dichlorvos-mediated neurotoxicity in rats by suppressing oxidative stress, acetylcholinesterase inhibition, and NF-κB-p65/p53/caspase-3-mediated apoptosis

BackgroundIn third-world countries, poisoning due to dichlorvos (DDVP), an organophosphate insecticide, is prevalent due to its widespread usage in household and agriculture, with the brain bearing the brunt of the consequences. Hence, this study assessed the likely beneficial impact of hesperidoside (HESP) on the DDVP-mediated cerebral dysfunction in rat model. MethodRandomization was employed to earmark forty-two rats into seven groups: control, DDVP alone (8 mg.kg⁻¹day⁻¹), DDVP plus HESP (50 and 100 mg.kg⁻¹day⁻¹) and reference drug atropine (0.2 mg.kg⁻¹day⁻¹), and HESP alone (50 and 100 mg.kg⁻¹day⁻¹). ResultsHESP intervention remarkably (p < 0.05) mitigated DDVP-potentiated augmentations in cerebral concentrations of H₂O₂, NO, and malondialdehyde; impaired DDVP-induced decrease in cerebral GSH, GST, SOD, catalase, glutathione peroxidase, and acetylcholinesterase; significantly (p < 0.05) suppressed DDVP-invoked upregulation of mRNA expression of NF-κB-p65, p53, BAX, caspase-3, and TNF-α; and significantly (p < 0.05) revoked DDVP-incited downregulation of interleukin-10. ConclusionHESP chemotherapeutic interventions enhanced cerebral functions in DDVP-treated rats by abrogating oxidative stress, acetylcholinesterase inhibition, and NF-κB-p65/p53/caspases-3 signaling.

Dawn-to-dusk intermittent fasting is associated with overexpression of autophagy genes: A prospective study on overweight and obese cohort

Aim and BackgroundA growing body of evidence supports the impact of intermittent fasting (IF) on longevity and healthy aging via the modulation of autophagy genes. The activation of the catabolic autophagic machinery (LAMP2, LC3B, ATG5, and ATG4D) has protective effects against degenerative aging and chronic diseases. This research examined the changes in the expression of the aforementioned genes upon the observance of dawn-to-dusk IF among metabolically healthy participants with overweight and obesity. MethodsFifty-one (51) participants (36 males and 15 females, 38.84 ± 11.73 years) with overweight and obesity (BMI = 29.75 ± 5.04 kg/m2) were recruited and monitored before and at the end of the commencement of the four-week IF. Six healthy subjects with normal BMI (21.4±2.20 kg/m2) were recruited only to standardize the reference for normal levels of gene expressions. At the two time points, anthropometric, biochemical, and dietary assessments were performed, and LAMP2, LC3B, ATG5, and ATG4D gene expressions were assessed using qRT-PCR on RNA extracted from whole blood samples. ResultsAt the end of IF, and compared to the pre-fasting levels, the relative gene expressions among participants with overweight/obesity were significantly increased for the three autophagy genes LAMP2, LC3B, and ATG5, with increments of about 4.2 folds, 1.9-fold, and 1.4-fold, respectively. In contrast, the increase in the ATG4D gene was not significant. Concomitantly, significant decreases were found in body weight, BMI, fat mass, body fat percent, hip and waist circumferences, LDL, IL-6, and TNF-a (P <0.05), While HDL, IL-10, and CD163 significantly increased (P <0.05). Binary logistic regression analysis for genetic expressions showed no significant association between high-energy intake, waist circumference, or obesity and the four gene expressions. ConclusionsFour consecutive weeks of dawn-to-dusk IF of Ramadan is associated with the upregulation of autophagy gene expressions in participants with overweight/obesity, and this may explain, at least in part, its favorable short-term temporal metabolic and health-improving effects on early aging-related markers. Hence, IF presumably may entail a protective impact against early markers of aging and metabolic diseases in participants with overweight/obesity.

Transcriptomic insight into the underlying mechanism of induced molting on reproductive remodeling, performance and egg quality in laying hen.

This study aimed to clarify the reproductive remodeling mechanism in enhancing production performance and egg quality during the fasting-induced molting process of laying hens. A total of two-hundred and forty 380-days-old Jingfen No. 6 laying hens, with an average laying rate of 78% were divided into four replicates, with 60 hens in each replicate to receive a four-stage molt induction experiment. The stages encompassed the pre-molt stage (T1), the molt stage (T2), the recovery stage (T3), and the second peak-laying stage (T4). The egg-laying rate and egg quality were recorded during all stages, and sample collection (serum, magnum of oviduct, ovary) was conducted at the end of each stage. The length and index of oviduct, the number of hierarchical follicles, and serum reproductive hormone levels were further measured, followed by the transcriptomic sequencing process on the magnum of the oviduct and ovarian tissues at each stage. Results showed that the fasting treatment induced atrophy of the oviducts, the disappearance of large yellow follicles in the ovaries, and the decrease in serum reproductive hormone levels compared to the pre-molt stage. Transcriptomic analysis revealed that differentially expressed genes were notably enriched in cytokine-cytokine receptor interactions, cell adhesion molecules, and the arachidonic acid metabolism signaling pathway during the remodeling phases of oviduct and ovary tissues. Key candidate genes such as BMPR1B, NEGR1, VTN, and CHAD emerged as pivotal in influencing reproductive function remodeling in molt-treated chickens. Additionally, genes associated with steroid biosynthesis showed significant up-regulation in the ovaries of molted hens, correlating positively with egg-laying rates. Furthermore, genes related to collagen and laminin displayed significant positive associations with Albumen height and Haugh unit values. The results indicate that fasting interventions might modulate the remodeling of reproductive functions in laying hens by altering cytokine-cytokine receptor interactions, cell adhesion molecules, and arachidonic acid metabolic pathways. Enhanced ovarian steroid biosynthesis and up-regulation of gene expression, including oviductal collagens post-molting, are crucial for enhancing laying rates and egg quality. These findings could offer novel thinking for refining molting protocols.

RhFGF21 protected PC12 cells against mitochondrial apoptosis triggered by H2O2 via the AKT-mediated ROS signaling pathway.

One of the pathological mechanisms of neurodegenerative diseases is that oxidative stress damages neurons. Therefore, reducing reactive oxygen species (ROS) overload may be a promising approach for preventing and treating neurological diseases. Fibroblast growth factor 21 (FGF21) is crucial for protecting and restoring various forms of pathological injury. Consequently, the operating mechanism of FGF21 was investigated. Our research revealed that rhFGF21 could enhance the cell viability by alleviating the damage to PC12cells after H2O2 action of via mechanisms decreasing mitochondrial apoptosis, reducing ROS production, increasing antioxidant enzyme levels, adenosine triphosphate (ATP) synthesis and mitochondrial membrane potential (MMP). Excessive ROS trigger cell apoptosis. Our findings revealed that tBHP counteracted the cell viability-boosting effect of rhFGF21 in H2O2-stimulated PC12cells, whereas N-acetyl-L-cysteine (NAC) enhanced the viability-promoting effect of rhFGF21 in these cells. AKT is crucial in mediating ROS-induced cell apoptosis. The treatment of PC12cells exposed to H2O2 with rhFGF21 resulted in upregulation of p-AKT expression. Moreover, rhFGF21 inhibited ROS levels and increased the cell viability, which were both reversed by administration of an AKT inhibitor (wortmannin). The research discovered that rhFGF21 mitigated mitochondrial apoptosis in PC12cells exposed to H2O2 through the functioning of the AKT and ROS signaling axis.

GPX3 Overexpression Ameliorates Cardiac Injury Post Myocardial Infarction Through Activating LSD1/Hif1α Axis.

Myocardial infarction (MI) often results in significant loss of cardiomyocytes (CMs), contributing to adverse ventricular remodelling and heart failure. Therefore, promoting CM survival during the acute stage of MI is crucial. This study aimed to investigate the potential role of GPX3 in cardiac repair following MI. First, plasma GPX3 levels were measured in patients with acute MI (AMI), and myocardial GPX3 expression was assessed in a mouse MI model. Furthermore, the effects of GPX3 on MI were investigated through CM-specific overexpression or knockdown invitro and invivo models. RNA sequencing and subsequent experiments were performed to uncover the molecular mechanisms underlying GPX3-related effects. Multi-omics database analysis and experimental verification revealed a significant upregulation of GPX3 expression in ischemic myocardium following MI and in CMs exposed to oxygen-glucose deprivation (OGD). Immunofluorescence results further confirmed elevated cytoplasmic GPX3 expression in CMs under hypoxic conditions. Invitro, GPX3 overexpression mitigated reactive oxygen species (ROS) production and enhanced CM survival during hypoxia, while GPX3 knockdown inhibited these processes. Invivo, CM-specific GPX3 overexpression in the infarct border zone significantly attenuated CM apoptosis and alleviated myocardial injury, promoting cardiac repair and long-term functional recovery. Mechanistically, GPX3 overexpression upregulated LSD1 and Hif1α protein expression, and rescue experiments confirmed the involvement of the LSD1/Hif1α pathway in mediating the protective effects of GPX3. Overall, our findings suggest that GPX3 exerts a protective role in ischemic myocardium post-MI, at least partially through the LSD1/Hif1α axis, highlighting its potential as a therapeutic target for MI treatment.

The Impact of miR-141 Overexpression on Apoptosis and Proliferation in Tscca and Tca8113 Cell Lines.

This study investigates the effect of miR-141 overexpression on the proliferation and apoptosis of tongue squamous cell carcinoma (TSCC) cell lines (Tscca and Tca8113). TSCC cells were stably transfected with a miR-141 lentivirus (Ubi-miR-141-SV40-EGFP-IRES-puromycin, miR-141 group) or an empty vector (Ubi-MCS-SV40-EGFP-IRES-puromycin, miR-NC group), with untreated cells serving as the Blank group. miR-141 expression was evaluated by rt-qPCR, transfection efficiency and apoptosis were assessed by flow cytometry, and cell proliferation was measured using the CCK-8 assay. Western blot and rt-qPCR were used to analyze the protein and mRNA expression of phosphatase and tensin homolog (PTEN) and B-cell lymphoma-2 (BCL2). Results: Flow cytometry confirmed successful establishment of stably transfected cell lines. The CCK-8 assay showed no significant difference in proliferation between the miR-NC and Blank groups (P > 0.05), while proliferation was significantly reduced in the miR-141 group. Apoptosis was significantly higher in the miR-141 group compared to both the miR-NC and Blank groups. Western blot and rt-qPCR analyses revealed upregulation of pten gene and PTEN protein expression, as well as downregulation of bcl2 gene and BCL2 protein expression in the miR-141 group relative to the miR-NC and Blank groups. Conclusion: In TSCC cells with high miR-141 expression, pten gene and PTEN protein levels were elevated, while bcl2 gene and BCL2 protein levels were reduced. These findings indicate that miR-141 may regulate the expression of PTEN and BCL2 at both the gene and protein levels, thereby influencing cell proliferation, migration, and apoptosis. To our knowledge, this is the first study to explore the effects of miR-141 transfection in TSCC, providing valuable insights into its role in tumor progression and potential regulatory targets.

Dissecting the role of pheromone-binding protein 2 in courtship behavior of male Grapholita molesta moths.

For fruit-boring pests like Grapholita molesta, the courtship behavior of male moths is a crucial aspect that can be utilized for developing control methods. This study investigates the role of pheromone-binding proteins (PBPs) in the courtship behavior of male G. molesta moths, particularly how sex pheromone components affect PBP expression. We found that three GmolPBPs genes were mainly expressed in the antennae of adult males, with no expression in larvae or pupae. Notably, GmolPBP2 expression was significantly higher in males, while GmolPBP1 was more pronounced in females. Unmated males exposed to females or the sex pheromone component Z8-12:Ac showed marked upregulation of GmolPBPs expression. Using affinity chromatography, we purified GmolPBPs and confirmed their binding affinities. Especially for GmolPBP2, this protein had a high affinity for Z8-12:Ac and E8-12:Ac. RNA interference targeting GmolPBPs revealed that silencing GmolPBP2 significantly reduced male responses to sex pheromone components and courtship behavior towards females, while silencing GmolPBP1 and GmolPBP3 had no significant effects. These findings highlight the crucial role of GmolPBP2 in mediating male courtship behavior, emphasizing the importance of PBPs in pheromone communication and mating success in G. molesta.

Tideglusib enhances ALP activity and upregulates RANKL expression in Osteoblast-macrophage Co-cultures within a 3D collagen scaffold.

Tideglusib (Tx) is known for its osteogenic potential, yet its effects on the interplay between osteoblasts and M1 macrophages remain underexplored. This in vitro study aimed to isolate and evaluate both the individual and combined roles of M1 macrophages and osteoblasts in macrophage differentiation and osteoblast function, specifically focusing on how these interactions influence protein expression of osteogenesis and osteoclastogenesis in the presence or absence of Tx. Osteoblast and macrophage cells were co-cultured in direct contact for 24 and 48 h, with or without the presence of Tx. ALP activity, the expression of inflammatory-related genes using RT-qPCR, and histological analyses were performed. Co-culturing osteoblasts and M1 macrophages with Tx increased alkaline phosphatase production, indicative of enhanced osteoblast activity. Histological assessments revealed that Tx treatment contributed to the stability and maintenance of cell morphology within the scaffold, suggesting a supportive environment for cell viability and function. Tx significantly reduced the expression of pro-inflammatory markers, such as TNF-α and IL-1β, in the co-culture at both 24 and 48 h Tx also effectively inhibited osteoclastogenic differentiation in macrophages, thereby diminishing their pro-inflammatory phenotype. Tx increased ALP activity and produced a significant up-regulation of RANKL expression, indicating enhanced osteoblast differentiation and osteoclast activation. Tx mitigates macrophage-driven inflammation. Tx may enhance bone regeneration by modulating inflammatory responses and preserving cell integrity.

Estrogen Promotes the Proliferation and Migration of Endometrial Cancer Through the GPER-Mediated NOTCH Pathway.

This study aims to investigate the expression of GPER in EC, assess the impact of estrogen on the proliferation and migration of EC via GPER, and examine the potential role of GPER in mediating the NOTCH pathway to influence EC proliferation and migration. The expression of GPER and its correlation with clinicopathological features were investigated using clinical data. Cell proliferation was assessed through MTT and EdU assays, while cell migration ability was evaluated using wound healing and transwell assays. Western blot analysis was conducted to detect proteins associated with the GPER and NOTCH signaling pathways. Additionally, xenograft tumor models were established to investigate the potential role of estrogen in mediating the NOTCH pathway via GPER. The results demonstrated a significant upregulation of GPER expression in EC, which was associated with clinical stage and metastasis. In vitro experiments provided evidence that estrogen promotes EC cell proliferation and metastasis by enhancing the expression levels of GPER, Notch1, and Hes-1 proteins. Conversely, knocking down or suppressing GPER effectively reverses these effects. Furthermore, treatment with JAG-1, an agonist for the NOTCH pathway, counteracts si-GPER's inhibitory impact on both proliferation and migration abilities of EC cells while increasing Notch1 and Hes-1 protein expression levels; however, it does not alter GPER expression. In vivo experiments have substantiated that estrogen facilitates EC proliferation via the GPER-mediated NOTCH pathway.

Antioxidant Peptides from Miiuy Croaker Swim Bladders: Ameliorating Effect and Mechanism in NAFLD Cell Model through Regulation of Hypolipidemic and Antioxidant Capacity

In this work, the hypolipidemic and antioxidative capacity of FSGLR (S7) and GIEWA (S10) from miiuy croaker swim bladders was explored systematically in an oleic acid (OA)-induced nonalcoholic fatty liver disease (NAFLD) model of HepG2 cells. Moreover, the hypolipidemic activity of S7 and S10 and their antioxidative abilities were preliminarily investigated in combination with molecular docking technology. The results indicated that S7 and S10 could decrease the amount of lipid accumulation and the content of triglycerides (TG) and total cholesterol (TC) in the OA-induced NAFLD cell model in a dose-dependent manner. In addition, S7 and S10 exhibited better bile salt binding, pancreatic lipase (PL) inhibition, and cholesterol esterase (CE) inhibition capacities. The hypolipidemic mechanisms of S7 and S10 were connected with the downregulation of the mRNA expression levels of adipogenic factors, including sterol-regulatory element-binding protein-1c (SREBP-1c), acetyl-CoA carboxylase (ACC), sterol-regulatory element-binding protein (SREBP)-2, hydroxymethylglutaryl-CoA reductase (HMGR), and fatty acid synthase (FAS) (p &lt; 0.01), and the upregulation of the mRNA expression of β-oxidation-related factors, including carnitine palmitoyltransferase 1 (CPT-1), acyl-CoA oxidase 1 (ACOX-1), and peroxisome proliferator-activated receptor α (PPARα). Moreover, FSGLR (S7) and GIEWA (S10) could significantly protect HepG2 cells against OA-induced oxidative damage, and their antioxidant mechanisms were related to the increased activity of intracellular antioxidant proteases (superoxide dismutase, SOD; glutathione peroxidase, GSH-PX; catalase, CAT) to remove excess reactive oxygen species (ROS) and decrease the production of malondialdehyde (MDA). The presented findings indicate that the hypolipidemic and antioxidant functions and mechanisms of S7 and S10 could make them potential hypolipidemic and antioxidant candidates for the treatment of NAFLD.