1  6456 130 THYMOSIN BETA4 PREVENTS OXIDATIVE STRESS, INFLAMMATION, AND FIBROSIS IN ETHANOL- AND LPS-INDUCED LIVER INJURY IN MICE. THYMOSIN BETA 4 (TBETA4), AN ACTIN-SEQUESTERING PROTEIN, IS INVOLVED IN TISSUE DEVELOPMENT AND REGENERATION. IT PREVENTS INFLAMMATION AND FIBROSIS IN SEVERAL TISSUES. WE INVESTIGATED THE ROLE OF TBETA4 IN CHRONIC ETHANOL- AND ACUTE LIPOPOLYSACCHARIDE- (LPS-) INDUCED MOUSE LIVER INJURY. C57BL/6 MICE WERE FED 5% ETHANOL IN LIQUID DIET FOR 4 WEEKS PLUS BINGE ETHANOL (5 G/KG, GAVAGE) WITH OR WITHOUT LPS (2 MG/KG, INTRAPERITONEAL) FOR 6 HOURS. TBETA4 (1 MG/KG, INTRAPERITONEAL) WAS ADMINISTERED FOR 1 WEEK. WE DEMONSTRATED THAT TBETA4 PREVENTED ETHANOL- AND LPS-MEDIATED INCREASE IN LIVER INJURY MARKERS AS WELL AS CHANGES IN LIVER PATHOLOGY. IT ALSO PREVENTED ETHANOL- AND LPS-MEDIATED INCREASE IN OXIDATIVE STRESS BY DECREASING ROS AND LIPID PEROXIDATION AND INCREASING THE ANTIOXIDANTS, REDUCED GLUTATHIONE AND MANGANESE-DEPENDENT SUPEROXIDE DISMUTASE. IT ALSO PREVENTED THE ACTIVATION OF NUCLEAR FACTOR KAPPA B BY BLOCKING THE PHOSPHORYLATION OF THE INHIBITORY PROTEIN, IKAPPAB, THEREBY PREVENTED PROINFLAMMATORY CYTOKINE PRODUCTION. MOREOVER, TBETA4 PREVENTED FIBROGENESIS BY SUPPRESSING THE EPIGENETIC REPRESSOR, METHYL-CPG-BINDING PROTEIN 2, THAT COORDINATELY REVERSED THE EXPRESSION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA AND DOWNREGULATED FIBROGENIC GENES, PLATELET-DERIVED GROWTH FACTOR-BETA RECEPTOR, ALPHA-SMOOTH MUSCLE ACTIN, COLLAGEN 1, AND FIBRONECTIN, RESULTING IN REDUCED FIBROSIS. OUR DATA SUGGEST THAT TBETA4 HAS ANTIOXIDANT, ANTI-INFLAMMATORY, AND ANTIFIBROTIC POTENTIAL DURING ALCOHOLIC LIVER INJURY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
2  5714  24 SIRT3 OVEREXPRESSION AND EPIGENETIC SILENCING OF CATALASE REGULATE ROS ACCUMULATION IN CLL CELLS ACTIVATING AXL SIGNALING AXIS. MITOCHONDRIAL METABOLISM IS THE KEY SOURCE FOR ABUNDANT ROS IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) CELLS. HERE, WE DETECTED SIGNIFICANTLY LOWER SUPEROXIDE ANION (O(2)(-)) LEVELS WITH INCREASED ACCUMULATION OF HYDROGEN PEROXIDE (H(2)O(2)) IN CLL CELLS VS. NORMAL B-CELLS. FURTHER ANALYSIS INDICATED THAT MITOCHONDRIAL SUPEROXIDE DISMUTASE (SOD)2, WHICH CONVERTS O(2)(-) INTO H(2)O(2) REMAINED DEACETYLATED IN CLL CELLS DUE TO SIRT3 OVEREXPRESSION RESULTING ITS CONSTITUTIVE ACTIVATION. IN ADDITION, CATALASE EXPRESSION WAS ALSO REDUCED IN CLL CELLS SUGGESTING IMPAIRMENT OF H(2)O(2)-CONVERSION INTO WATER AND O(2) WHICH MAY CAUSE H(2)O(2)-ACCUMULATION. IMPORTANTLY, WE IDENTIFIED TWO CPG-ISLANDS IN THE CATALASE PROMOTER AND DISCOVERED THAT WHILE THE DISTAL CPG-ISLAND (-3619 TO -3765) REMAINED METHYLATED IN BOTH NORMAL B-CELLS AND CLL CELLS, VARIABLE DEGREES OF METHYLATION WERE DISCERNIBLE IN THE PROXIMAL CPG-ISLAND (-174 TO -332) ONLY IN CLL CELLS. FINALLY, TREATMENT OF CLL CELLS WITH A DEMETHYLATING AGENT INCREASED CATALASE MRNA LEVELS. FUNCTIONALLY, ROS ACCUMULATION IN CLL CELLS ACTIVATED THE AXL SURVIVAL AXIS WHILE UPREGULATED SIRT3, SUGGESTING THAT CLL CELLS RAPIDLY REMOVE HIGHLY REACTIVE O(2)(-) TO AVOID ITS CYTOTOXIC EFFECT BUT MAINTAIN INCREASED H(2)O(2)-LEVEL TO PROMOTE CELL SURVIVAL. THEREFORE, ABROGATION OF ABERRANTLY ACTIVATED CELL SURVIVAL PATHWAYS USING ANTIOXIDANTS CAN BE AN EFFECTIVE INTERVENTION IN CLL THERAPY IN COMBINATION WITH CONVENTIONAL AGENTS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
3   603  39 BETULINIC ACID INHIBITS ENDOMETRIOSIS THROUGH SUPPRESSION OF ESTROGEN RECEPTOR BETA SIGNALING PATHWAY. ENDOMETRIOSIS IS AN INFLAMMATORY GYNECOLOGICAL DISORDER CHARACTERIZED BY ENDOMETRIAL TISSUE GROWTH LOCATED OUTSIDE OF THE UTERINE CAVITY IN ADDITION TO CHRONIC PELVIC PAIN AND INFERTILITY. IN THIS STUDY, WE AIM TO DEVELOP A POTENTIAL THERAPEUTIC TREATMENT BASED ON THE PATHOGENESIS AND MECHANISM OF ENDOMETRIOSIS. OUR PRELIMINARY DATA SHOWED THAT THE EXPRESSION OF ESTROGEN RECEPTOR BETA (ERBETA) WAS SIGNIFICANTLY INCREASED, WHILE ERALPHA WAS SIGNIFICANTLY DECREASED, IN ENDOMETRIOTIC CELLS COMPARED TO NORMAL ENDOMETRIAL CELLS. FURTHER INVESTIGATION SHOWED THAT BETULINIC ACID (BA) TREATMENT SUPPRESSED ERBETA EXPRESSION THROUGH EPIGENETIC MODIFICATION ON THE ERBETA PROMOTER, WHILE HAD NO EFFECT ON ERALPHA EXPRESSION. IN ADDITION, BA TREATMENT SUPPRESSES ERBETA TARGET GENES, INCLUDING SUPEROXIDE DISMUTASE 2 (SOD2), NUCLEAR RESPIRATORY FACTOR-1 (NRF1), CYCLOOXYGENASE 2 (COX2), AND MATRIX METALLOPROTEINASE-1 (MMP1), SUBSEQUENTLY INCREASING OXIDATIVE STRESS, TRIGGERING MITOCHONDRIAL DYSFUNCTION, DECREASING ELEVATED PROINFLAMMATORY CYTOKINES, AND EVENTUALLY SUPPRESSING ENDOMETRIOTIC CELL PROLIFERATION, MIMICKING THE EFFECT OF ERBETA KNOCKDOWN. ON THE OTHER HAND, GAIN OF ERBETA BY LENTIVIRUS INFECTION IN NORMAL ENDOMETRIAL CELLS RESULTED IN INCREASED CELL PROLIFERATION AND PROINFLAMMATORY CYTOKINE RELEASE, WHILE BA TREATMENT DIMINISHED THIS EFFECT THROUGH ERBETA SUPPRESSION WITH SUBSEQUENT OXIDATIVE STRESS AND APOPTOSIS. OUR RESULTS INDICATE THAT ERBETA MAY BE A MAJOR DRIVING FORCE FOR THE DEVELOPMENT OF ENDOMETRIOSIS, WHILE BA INHIBITS ENDOMETRIOSIS THROUGH SPECIFIC SUPPRESSION OF THE ERBETA SIGNALING PATHWAY. THIS STUDY PROVIDES A NOVEL THERAPEUTIC STRATEGY FOR ENDOMETRIOSIS TREATMENT THROUGH BA-MEDIATED ERBETA SUPPRESSION.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
4  5479  25 RESVERATROL ATTENUATES CIGARETTE SMOKE EXTRACT INDUCED CELLULAR SENESCENCE IN HUMAN AIRWAY EPITHELIAL CELLS BY REGULATING THE MIR-34A/SIRT1/NF-KAPPAB PATHWAY. CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS CHARACTERIZED BY ACCELERATED LUNG AGING. SMOKING IS THE CRITICAL RISK FACTOR FOR COPD. CELLULAR SENESCENCE OF AIRWAY EPITHELIAL CELLS IS THE CYTOLOGICAL BASIS OF ACCELERATED LUNG AGING IN COPD, AND THE REGULATION OF MICRORNAS (MIRNAS) IS THE CENTRAL EPIGENETIC MECHANISM OF CELLULAR SENESCENCE. RESVERATROL (RES) IS A POLYPHENOL WITH ANTI-AGING PROPERTIES. THIS STUDY INVESTIGATED WHETHER RES ATTENUATES CIGARETTE SMOKE EXTRACT (CSE)-INDUCED CELLULAR SENESCENCE IN HUMAN AIRWAY EPITHELIAL CELLS (BEAS-2B) THROUGH THE MIR-34A/SIRT1/NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) PATHWAY. BEAS-2B CELLS WERE TREATED WITH RES, CSE AND TRANSFECTED WITH MIR-34A-5P MIMICS. CELLULAR SENESCENCE WAS EVALUATED BY SENESCENCE -RELATED BETA-GALACTOSIDASE (SA-BETA-GAL) STAINING AND EXPRESSION OF SENESCENCE-RELATED GENES (P16, P21, AND P53). THE EXPRESSIONS OF MIR-34A-5P, SIRT1, AND NF-KAPPAB P65 WERE EXAMINED USING QUANTITATIVE REAL TIME POLYMERASE CHAIN REACTION AND WESTERN BLOTTING. THE SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP) CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) WERE ASSESSED BY ENZYME-LINKED IMMUNOSORBENT ASSAY. THE BINDING BETWEEN MIR-34A-5P AND SIRT1 WAS CONFIRMED BY DUAL-LUCIFERASE REPORTER ASSAY. THE RESULTS SHOWED THAT CSE DOSE-DEPENDENTLY DECREASED CELL VIABILITY AND ELEVATED CELLULAR SENESCENCE, CHARACTERIZED BY INCREASED SA-BETA-GAL STAINING AND SENESCENCE-RELATED GENE EXPRESSIONS (P16, P21, AND P53). FURTHER, CSE DOSE-DEPENDENTLY INCREASED THE EXPRESSION OF MIR-34A-5P AND SASP CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) IN BEAS-2B CELLS. PRETREATMENT WITH RES INHIBITED CSE-INDUCED CELLULAR SENESCENCE AND SECRETION OF SASP CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) IN A DOSE-DEPENDENT MANNER. MOREOVER, RES REVERSED THE CSE-INDUCED DOWN-REGULATION OF SIRT1 AND UP-REGULATION OF MIR-34A-5P AND NF-KAPPAB P65. SIRT1 IS A TARGET OF MIR-34A-5P. OVEREXPRESSION OF MIR-34A-5P VIA TRANSFECTION WITH MIR-34A-5P MIMIC IN BEAS-2B CELLS ATTENUATED THE INHIBITORY EFFECT OF RES ON CELLULAR SENESCENCE, ACCOMPANIED BY REVERSING THE EXPRESSION OF SIRT1 AND NF-KAPPAB P65. IN CONCLUSION, RES ATTENUATED CSE-INDUCED CELLULAR SENESCENCE IN BEAS-2B CELLS BY REGULATING THE MIR-34A/SIRT1/NF-KAPPAB PATHWAY, WHICH MAY PROVIDE A NEW APPROACH FOR COPD TREATMENT.	2022	
                                                                                                                                                                                                                           
5  2774  44 EXTRACELLULAR SUPEROXIDE DISMUTASE (EC-SOD) REGULATES GENE METHYLATION AND CARDIAC FIBROSIS DURING CHRONIC HYPOXIC STRESS. CHRONIC HYPOXIC STRESS INDUCES EPIGENETIC MODIFICATIONS MAINLY DNA METHYLATION IN CARDIAC FIBROBLASTS, INACTIVATING TUMOR SUPPRESSOR GENES (RASSF1A) AND ACTIVATING KINASES (ERK1/2) LEADING TO FIBROBLAST PROLIFERATION AND CARDIAC FIBROSIS. THE RAS/ERK SIGNALING PATHWAY IS AN INTRACELLULAR SIGNAL TRANSDUCTION CRITICALLY INVOLVED IN FIBROBLAST PROLIFERATION. RASSF1A FUNCTIONS THROUGH ITS EFFECT ON DOWNSTREAM ERK1/2. THE ANTIOXIDANT ENZYME, EXTRACELLULAR SUPEROXIDE DISMUTASE (EC-SOD), DECREASES OXIDATIVE STRESS FROM CHRONIC HYPOXIA, BUT ITS EFFECTS ON THESE EPIGENETIC CHANGES HAVE NOT BEEN FULLY EXPLORED. TO TEST OUR HYPOTHESIS, WE USED AN IN-VITRO MODEL: WILD-TYPE C57B6 MALE MICE (WT) AND TRANSGENIC MALES WITH AN EXTRA COPY OF HUMAN HEC-SOD (TG). THE STUDIED ANIMALS WERE HOUSED IN HYPOXIA (10% O(2)) FOR 21 DAYS. THE RIGHT VENTRICULAR TISSUE WAS STUDIED FOR CARDIAC FIBROSIS MARKERS USING RT-PCR AND WESTERN BLOT ANALYSES. PRIMARY C57BL6 MOUSE CARDIAC FIBROBLAST TISSUE CULTURE WAS USED TO STUDY THE IN-VITRO MODEL, THE DOWNSTREAM EFFECTS OF RASSF-1 EXPRESSION AND METHYLATION, AND ITS RELATION TO ERK1/2. OUR FINDINGS SHOWED A SIGNIFICANT INCREASE IN CARDIAC FIBROSIS MARKERS: COLLAGEN 1, ALPHA SMOOTH MUSCLE ACTIN (ASMA), AND SNAIL, IN THE WT HYPOXIC ANIMALS AS COMPARED TO THE TG HYPOXIC GROUP (P < 0.05). THE EXPRESSION OF DNA METHYLATION ENZYMES (DNMT 1&3B) WAS SIGNIFICANTLY INCREASED IN THE WT HYPOXIC MICE AS COMPARED TO THE HYPOXIC TG MICE (P < 0.001). RASSF1A EXPRESSION WAS SIGNIFICANTLY LOWER AND ERK1/2 WAS SIGNIFICANTLY HIGHER IN HYPOXIA WT COMPARED TO THE HYPOXIC TG GROUP (P < 0.05). USE OF SIRNA TO BLOCK RASSF1A GENE EXPRESSION IN MURINE CARDIAC FIBROBLAST TISSUE CULTURE LED TO INCREASED FIBROBLAST PROLIFERATION (P < 0.05). METHYLATION OF THE RASSF1A PROMOTER REGION WAS SIGNIFICANTLY REDUCED IN THE TG HYPOXIC GROUP COMPARED TO THE WT HYPOXIC GROUP (0.59 VS. 0.75, RESPECTIVELY). BASED ON OUR FINDINGS, WE CAN SPECULATE THAT EC-SOD SIGNIFICANTLY ATTENUATES RASSF1A GENE METHYLATION AND CAN ALLEVIATE CARDIAC FIBROSIS INDUCED BY HYPOXIA.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
6  6298  42 THE PROTECTIVE EFFECT OF ZEBULARINE, AN INHIBITOR OF DNA METHYLTRANSFERASE, ON RENAL TUBULOINTERSTITIAL INFLAMMATION AND FIBROSIS. RENAL FIBROSIS, THE FINAL PATHWAY OF CHRONIC KIDNEY DISEASE, IS CAUSED BY GENETIC AND EPIGENETIC MECHANISMS. ALTHOUGH DNA METHYLATION HAS DRAWN ATTENTION AS A DEVELOPING MECHANISM OF RENAL FIBROSIS, ITS CONTRIBUTION TO RENAL FIBROSIS HAS NOT BEEN CLARIFIED. TO ADDRESS THIS ISSUE, THE EFFECT OF ZEBULARINE, A DNA METHYLTRANSFERASE INHIBITOR, ON RENAL INFLAMMATION AND FIBROSIS IN THE MURINE UNILATERAL URETERAL OBSTRUCTION (UUO) MODEL WAS ANALYZED. ZEBULARINE SIGNIFICANTLY ATTENUATED RENAL TUBULOINTERSTITIAL FIBROSIS AND INFLAMMATION. ZEBULARINE DECREASED TRICHROME, ALPHA-SMOOTH MUSCLE ACTIN, COLLAGEN IV, AND TRANSFORMING GROWTH FACTOR-BETA1 STAINING BY 56.2%. 21.3%, 30.3%, AND 29.9%, RESPECTIVELY, AT 3 DAYS, AND BY 54.6%, 41.9%, 45.9%, AND 61.7%, RESPECTIVELY, AT 7 DAYS AFTER UUO. ZEBULARINE DOWNREGULATED MRNA EXPRESSION LEVELS OF MATRIX METALLOPROTEINASE (MMP)-2, MMP-9, FIBRONECTIN, AND SNAIL1 BY 48.6%. 71.4%, 31.8%, AND 42.4%, RESPECTIVELY, AT 7 DAYS AFTER UUO. ZEBULARINE ALSO SUPPRESSED THE ACTIVATION OF NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND THE EXPRESSION OF PRO-INFLAMMATORY CYTOKINES, INCLUDING TUMOR NECROSIS FACTOR-ALPHA, INTERLEUKIN (IL)-1BETA, AND IL-6, BY 69.8%, 74.9%, AND 69.6%, RESPECTIVELY, IN OBSTRUCTED KIDNEYS. FURTHERMORE, INHIBITING DNA METHYLTRANSFERASE BUTTRESSED THE NUCLEAR EXPRESSION OF NUCLEAR FACTOR (ERYTHROID-DERIVED 2)-LIKE FACTOR 2, WHICH UPREGULATED DOWNSTREAM EFFECTORS SUCH AS CATALASE (1.838-FOLD INCREASE AT 7 DAYS, P < 0.01), SUPEROXIDE DISMUTASE 1 (1.494-FOLD INCREASE AT 7 DAYS, P < 0.05), AND NAD(P)H: QUINONE OXIDOREDUATE-1 (1.376-FOLD INCREASE AT 7 DAYS, P < 0.05) IN OBSTRUCTED KIDNEYS. COLLECTIVELY, THESE FINDINGS SUGGEST THAT INHIBITING DNA METHYLATION RESTORES THE DISRUPTED BALANCE BETWEEN PRO-INFLAMMATORY AND ANTI-INFLAMMATORY PATHWAYS TO ALLEVIATE RENAL INFLAMMATION AND FIBROSIS. THEREFORE, THESE RESULTS HIGHLIGHT THE POSSIBILITY OF DNA METHYLTRANSFERASES AS THERAPEUTIC TARGETS FOR TREATING RENAL INFLAMMATION AND FIBROSIS.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
7  5227  33 PRMT6 MEDIATES INFLAMMATION VIA ACTIVATION OF THE NF-KAPPAB/P65 PATHWAY ON A CIGARETTE SMOKE EXTRACT-INDUCED MURINE EMPHYSEMA MODEL. INTRODUCTION: SMOKE-DRIVEN LUNG INFLAMMATION IS CONSIDERED TO BE THE MAJOR PATHOPHYSIOLOGY MECHANISM OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD)/EMPHYSEMA. PROTEIN ARGININE METHYLTRANSFERASE 6 (PRMT6) IS A KEY EPIGENETIC ENZYME, WHICH IS RELATED TO PROTECTING THE TRI-METHYLATION OF H3K4 (H3K4ME3). WE HYPOTHESIZED THAT PTMT6 PROTECTS LUNG INFLAMMATION THROUGH THE NUCLEAR FACTOR KAPPA B (NF-KAPPAB) PATHWAY. METHODS: MICE WERE INJECTED WITH CIGARETTE SMOKE EXTRACT (CSE) OR PBS TO ESTABLISH A MICE MODEL, INTRATRACHEALLY INSTILLED WITH OVEREXPRESSED PRMT6 OR NEGATIVE CONTROL VECTOR. MORPHOMETRY OF LUNG SLIDES AND LUNG FUNCTION WERE MEASURED. WE DETERMINED THE PROTEIN EXPRESSION OF PRMT6 AND ITS RELATED HISTONE TARGETS, THE ACTIVATION OF NF-KAPPAB PATHWAY, THE LEVEL OF TUMOR NECROSIS FACTOR ALPHA (TNFALPHA) AND INTERLEUKIN-1BETA (IL-1BETA). RESULTS: AFTER PRMT6 OVEREXPRESSION, THE MORPHOMETRY INDEXES AND LUNG FUNCTION WERE IMPROVED. ALSO, THE EXPRESSION OF H3K4ME3 WAS DECREASED. OVEREXPRESSED PRMT6 COULD SUPPRESS CSE-INDUCED NF-KAPPAB ACTIVATION AND PRO-INFLAMMATION GENES EXPRESSION. CONCLUSIONS: THE OVEREXPRESSED PRMT6 COULD SERVE AS AN INFLAMMATION INHIBITOR, POTENTIALLY THROUGH BLOCKING THE NF-KAPPAB/P65 PATHWAY IN THE MURINE EMPHYSEMA MODEL.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
8  1826  38 EFFECTS OF HISTONE DEACETYLASE INHIBITOR ON EXTRACELLULAR MATRIX PRODUCTION IN HUMAN NASAL POLYP ORGAN CULTURES. BACKGROUND: NASAL POLYPOSIS IS ASSOCIATED WITH A CHRONIC INFLAMMATORY CONDITION OF THE SINONASAL MUCOSA AND INVOLVES MYOFIBROBLAST DIFFERENTIATION AND EXTRACELLULAR MATRIX (ECM) ACCUMULATION. EPIGENETIC MODULATION BY HISTONE DEACETYLASE (HDAC) INHIBITORS INCLUDING TRICHOSTATIN A (TSA) HAS BEEN REPORTED TO HAVE INHIBITORY EFFECTS ON MYOFIBROBLAST DIFFERENTIATION IN LUNG AND RENAL FIBROBLASTS. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE INHIBITORY EFFECT OF TSA ON MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION IN NASAL POLYP ORGAN CULTURES. METHODS: NASAL POLYP TISSUES FROM 18 PATIENTS WERE ACQUIRED DURING ENDOSCOPIC SINUS SURGERY. AFTER ORGAN CULTURE, NASAL POLYPS WERE STIMULATED WITH TGF-BETA1 AND THEN TREATED WITH TSA. ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), FIBRONECTIN, AND COLLAGEN TYPE I EXPRESSION LEVELS WERE EXAMINED BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (PCR), REAL-TIME PCR, WESTERN BLOT, AND IMMUNOFLUORESCENT STAINING. HDAC2, HDAC4, AND ACETYLATED H4 EXPRESSION LEVELS WERE ASSAYED BY WESTERN BLOT. CYTOTOXICITY WAS ANALYZED BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE BIOTIN-DUTP NICK END LABELING ASSAY. RESULTS: THE EXPRESSION LEVELS OF ALPHA-SMA, FIBRONECTIN, AND COLLAGEN TYPE 1 WERE INCREASED IN NASAL POLYP AFTER TRANSFORMING GROWTH FACTOR (TGF) BETA1 TREATMENT. TSA-INHIBITED TGF-BETA1 INDUCED THESE GENE AND PROTEIN EXPRESSION LEVELS. FURTHERMORE, TSA SUPPRESSED PROTEIN EXPRESSION LEVELS OF HDAC2 AND HDAC4. HOWEVER, TSA INDUCED HYPERACETYLATION OF HISTONES H4. TREATMENT WITH TGF-BETA1 WITH OR WITHOUT TSA DID NOT HAVE CYTOTOXIC EFFECT. CONCLUSION: THESE FINDINGS PROVIDE NOVEL INSIGHTS INTO THE EPIGENETIC REGULATION IN MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION OF NASAL POLYP. TSA COULD BE A CANDIDATE OF A THERAPEUTIC AGENT FOR REVERSING THE TGF-BETA1-INDUCED ECM SYNTHESIS THAT LEADS TO NASAL POLYP DEVELOPMENT.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
9  2278  29 EPIGENETIC REGULATION BY SUV4-20H1 IN CARDIOPULMONARY PROGENITOR CELLS IS REQUIRED TO PREVENT PULMONARY HYPERTENSION AND CHRONIC OBSTRUCTIVE PULMONARY DISEASE. BACKGROUND: THE PATHOGENESIS OF LIFE-THREATENING CARDIOPULMONARY DISEASES SUCH AS PULMONARY HYPERTENSION (PH) AND CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) ORIGINATES FROM A COMPLEX INTERPLAY OF ENVIRONMENTAL FACTORS AND GENETIC PREDISPOSITIONS THAT IS NOT FULLY UNDERSTOOD. LIKEWISE, LITTLE IS KNOWN ABOUT DEVELOPMENTAL ABNORMALITIES OR EPIGENETIC DYSREGULATIONS THAT MIGHT PREDISPOSE FOR PH OR COPD IN ADULT INDIVIDUALS. METHODS: TO IDENTIFY PATHOLOGY-ASSOCIATED EPIGENETIC ALTERATION IN DISEASED LUNG TISSUES, WE SCREENED A COHORT OF HUMAN PATIENTS WITH PH AND COPD FOR CHANGES OF HISTONE MODIFICATIONS BY IMMUNOFLUORESCENCE STAINING. TO ANALYZE THE FUNCTION OF H4K20ME2/3 IN LUNG PATHOGENESIS, WE DEVELOPED A SERIES OF SUV4-20H1 KNOCKOUT MOUSE LINES TARGETING CARDIOPULMONARY PROGENITOR CELLS AND DIFFERENT HEART AND LUNG CELL TYPES, FOLLOWED BY HEMODYNAMIC STUDIES AND MORPHOMETRIC ASSESSMENT OF TISSUE SAMPLES. MOLECULAR, CELLULAR, AND BIOCHEMICAL TECHNIQUES WERE APPLIED TO ANALYZE THE FUNCTION OF SUV4-20H1-DEPENDENT EPIGENETIC PROCESSES IN CARDIOPULMONARY PROGENITOR CELLS AND THEIR DERIVATIVES. RESULTS: WE DISCOVERED A STRONG REDUCTION OF THE HISTONE MODIFICATIONS OF H4K20ME2/3 IN HUMAN PATIENTS WITH COPD BUT NOT PATIENTS WITH PH THAT DEPEND ON THE ACTIVITY OF THE H4K20 DI-METHYLTRANSFERASE SUV4-20H1. LOSS OF SUV4-20H1 IN CARDIOPULMONARY PROGENITOR CELLS CAUSED A COPD-LIKE/PH PHENOTYPE IN MICE INCLUDING THE FORMATION OF PERIVASCULAR TERTIARY LYMPHOID TISSUE AND GOBLET CELL HYPERPLASIA, HYPERPROLIFERATION OF SMOOTH MUSCLE CELLS/MYOFIBROBLASTS, IMPAIRED ALVEOLARIZATION AND MATURATION DEFECTS OF THE MICROVASCULATURE LEADING TO MASSIVE RIGHT VENTRICULAR DILATATION AND PREMATURE DEATH. MECHANISTICALLY, SUV4-20H1 BINDS DIRECTLY TO THE 5'-UPSTREAM REGULATORY ELEMENT OF THE SUPEROXIDE DISMUTASE 3 (SOD3) GENE TO REPRESS ITS EXPRESSION. INCREASED LEVELS OF THE EXTRACELLULAR SOD3 ENZYME IN SUV4-20H1 MUTANTS INCREASES HYDROGEN PEROXIDE CONCENTRATIONS, CAUSING VASCULAR DEFECTS AND IMPAIRING ALVEOLARIZATION. CONCLUSIONS: OUR FINDINGS REVEAL A PIVOTAL ROLE OF THE HISTONE MODIFIER SUV4-20H1 IN CARDIOPULMONARY CODEVELOPMENT AND UNCOVER THE DEVELOPMENTAL ORIGINS OF CARDIOPULMONARY DISEASES. WE ASSUME THAT THE STUDY WILL FACILITATE THE UNDERSTANDING OF PATHOGENIC EVENTS CAUSING PH AND COPD AND AID THE DEVELOPMENT OF EPIGENETIC DRUGS FOR THE TREATMENT OF CARDIOPULMONARY DISEASES.	2021	
                                                                                                                                  
10  5868  35 SUPPRESSIVE EFFECTS OF METFORMIN ON T-HELPER 1-RELATED CHEMOKINES EXPRESSION IN THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1. PURPOSE OF THE STUDY: TYPE 1 AND TYPE 2 DIABETES MELLITUS (DM) ARE CHRONIC T-CELL-MEDIATED INFLAMMATORY DISEASES. METFORMIN IS A WIDELY USED DRUG FOR TYPE 2 DM THAT REDUCES THE NEED FOR INSULIN IN TYPE 1 DM. HOWEVER, WHETHER METFORMIN HAS AN ANTI-INFLAMMATORY EFFECT FOR TREATING DM IS UNKNOWN. WE INVESTIGATED THE ANTI-INFLAMMATORY MECHANISM OF METFORMIN IN THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1. MATERIALS AND METHODS: THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1 WAS PRETREATED WITH METFORMIN AND STIMULATED WITH LIPOPOLYSACCHARIDE (LPS). THE PRODUCTION OF T-HELPER (TH)-1-RELATED CHEMOKINES INCLUDING INTERFERON-GAMMA-INDUCED PROTEIN-10 (IP-10) AND MONOCYTE CHEMOATTRACTANT PROTEIN-1 (MCP-1), TH2-RELATED CHEMOKINE MACROPHAGE-DERIVED CHEMOKINE, AND THE PROINFLAMMATORY CHEMOKINE TUMOR NECROSIS FACTOR-ALPHA WAS MEASURED USING ENZYME-LINKED IMMUNOSORBENT ASSAY. INTRACELLULAR SIGNALING PATHWAYS WERE INVESTIGATED USING WESTERN BLOT ANALYSIS AND CHROMATIN IMMUNOPRECIPITATION ASSAY. RESULTS: METFORMIN SUPPRESSED LPS-INDUCED IP-10 AND MCP-1 PRODUCTION AS WELL AS LPS-INDUCED PHOSPHORYLATION OF C-JUN N-TERMINAL KINASE (JNK), P38, EXTRACELLULAR SIGNAL-REGULATED KINASE (ERK), AND NUCLEAR FACTOR-KAPPA B (NF-KAPPAB). MOREOVER, METFORMIN SUPPRESSED LPS-INDUCED ACETYLATION OF HISTONES H3 AND H4 AT THE IP-10 PROMOTER. CONCLUSIONS: METFORMIN SUPPRESSED THE PRODUCTION OF TH1-RELATED CHEMOKINES IP-10 AND MCP-1 IN THP-1 CELLS. SUPPRESSIVE EFFECTS OF METFORMIN ON IP-10 PRODUCTION MIGHT BE ATTRIBUTED AT LEAST PARTIALLY TO THE JNK, P38, ERK, AND NF-KAPPAB PATHWAYS AS WELL AS TO EPIGENETIC REGULATION THROUGH THE ACETYLATION OF HISTONES H3 AND H4. THESE RESULTS INDICATED THE THERAPEUTIC ANTI-INFLAMMATORY POTENTIAL OF METFORMIN.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
11  1848  35 EFFECTS OF VENLAFAXINE ON THE EXPRESSION LEVEL AND METHYLATION STATUS OF GENES INVOLVED IN OXIDATIVE STRESS IN RATS EXPOSED TO A CHRONIC MILD STRESS. RECENT HUMAN AND ANIMAL STUDIES INDICATE THAT OXIDATIVE AND NITROSATIVE STRESS MAY PLAY A ROLE IN THE AETIOLOGY AND PATHOGENESIS OF DEPRESSION. THIS STUDY INVESTIGATES THE EFFECT OF CHRONIC ADMINISTRATION OF THE SEROTONIN-NOREPINEPHRINE REUPTAKE INHIBITOR, VENLAFAXINE, ON THE EXPRESSION AND METHYLATION STATUS OF SOD1, SOD2, GPX1, GPX4, CAT, NOS1 AND NOS2 IN THE BRAIN AND BLOOD OF RATS EXPOSED TO A CHRONIC MILD STRESS (CMS) MODEL OF DEPRESSION. SEPARATE GROUPS OF ANIMALS WERE EXPOSED TO CMS FOR 2 OR 7 WEEKS; THE SECOND GROUP RECEIVED SALINE OR VENLAFAXINE (10 MG/KG/D, IP) FOR 5 WEEKS. AFTER COMPLETION OF BOTH STRESS CONDITIONS AND DRUG ADMINISTRATION, THE MRNA AND PROTEIN EXPRESSION OF SELECTED GENES AND THE METHYLATION STATUS OF THEIR PROMOTERS WERE MEASURED IN PERIPHERAL MONONUCLEAR BLOOD CELLS (PBMCS) AND IN BRAIN STRUCTURES (HIPPOCAMPUS, AMYGDALA, HYPOTHALAMUS, MIDBRAIN, CORTEX, BASAL GANGLIA) WITH THE USE OF TAQMAN GENE EXPRESSION ASSAY, WESTERN BLOT AND METHYLATION-SENSITIVE HIGH-RESOLUTION MELTING TECHNIQUES. CMS CAUSED A DECREASE IN SUCROSE CONSUMPTION, AND THIS EFFECT WAS NORMALIZED BY FLUOXETINE. IN PBMCS, SOD1, SOD2 AND NOS2 MRNA EXPRESSION CHANGED ONLY AFTER VENLAFAXINE ADMINISTRATION. IN BRAIN, CAT, GPX1, GPX4 AND NOS1 GENE EXPRESSION CHANGED FOLLOWING CMS OR VENLAFAXINE EXPOSURE, MOST PROMINENTLY IN THE HIPPOCAMPUS, MIDBRAIN AND BASAL GANGLIA. CMS INCREASED THE METHYLATION OF THE GPX1 PROMOTER IN PBMCS, THE SECOND GPX4 PROMOTER IN MIDBRAIN AND BASAL GANGLIA, AND SOD1 AND SOD2 IN HIPPOCAMPUS. THE CMS ANIMALS TREATED WITH VENLAFAXINE DISPLAYED A SIGNIFICANTLY HIGHER CAT LEVEL IN MIDBRAIN AND CEREBRAL CORTEX. CMS CAUSED AN ELEVATION OF GPX4 IN THE HIPPOCAMPUS, WHICH WAS LOWERED IN CEREBRAL CORTEX BY VENLAFAXINE. THE RESULTS INDICATE THAT CMS AND VENLAFAXINE ADMINISTRATION AFFECT THE METHYLATION OF PROMOTERS OF GENES INVOLVED IN OXIDATIVE AND NITROSATIVE STRESS. THEY ALSO INDICATE THAT PERIPHERAL AND CENTRAL TISSUE DIFFER IN THEIR RESPONSE TO STRESS OR ANTIDEPRESSANT TREATMENTS. IT IS POSSIBLE THAT THAT APART FROM DNA METHYLATION, A CRUCIAL ROLE OF EXPRESSION LEVEL OF GENES MAY BE PLAYED BY OTHER FORMS OF EPIGENETIC REGULATION, SUCH AS HISTONE MODIFICATION OR MICRORNA INTERFERENCE. THESE FINDINGS PROVIDE STRONG EVIDENCE FOR THESIS THAT ANALYSIS OF THE LEVEL OF MRNA AND PROTEIN EXPRESSION AS WELL AS THE STATUS OF PROMOTER METHYLATION CAN HELP IN UNDERSTANDING THE PATHOMECHANISMS OF MENTAL DISEASES, INCLUDING DEPRESSION, AND THE MECHANISMS OF ACTION OF DRUGS EFFECTIVE IN THEIR THERAPY.	2020	

12  1353  31 DEVELOPMENT AND CHARACTERIZATION OF A HYDROGEN PEROXIDE-RESISTANT CHOLANGIOCYTE CELL LINE: A NOVEL MODEL OF OXIDATIVE STRESS-RELATED CHOLANGIOCARCINOMA GENESIS. OXIDATIVE STRESS IS A CAUSE OF INFLAMMATION-RELATED DISEASES, INCLUDING CANCERS. CHOLANGIOCARCINOMA IS A LIVER CANCER WITH BILE DUCT EPITHELIAL CELL PHENOTYPES. OUR PREVIOUS STUDIES IN ANIMAL AND HUMAN MODELS INDICATED THAT OXIDATIVE STRESS IS A MAJOR CAUSE OF CHOLANGIOCARCINOMA DEVELOPMENT. HYDROGEN PEROXIDE (H2O2) CAN GENERATE HYDROXYL RADICALS, WHICH DAMAGE LIPIDS, PROTEINS, AND NUCLEIC ACIDS, LEADING TO CELL DEATH. HOWEVER, SOME CELLS CAN SURVIVE BY ADAPTING TO OXIDATIVE STRESS CONDITIONS, AND SELECTIVE CLONAL EXPANSION OF THESE RESISTANT CELLS WOULD BE INVOLVED IN OXIDATIVE STRESS-RELATED CARCINOGENESIS. THE PRESENT STUDY AIMED TO ESTABLISH H2O2-RESISTANT CELL LINE FROM AN IMMORTAL CHOLANGIOCYTE CELL LINE (MMNK1) BY CHRONIC TREATMENT WITH LOW-CONCENTRATION H2O2 (25 MUM). AFTER 72 DAYS OF INDUCTION, H2O2-RESISTANT CELL LINES (OX-MMNK1-L) WERE OBTAINED. THE OX-MMNK1-L CELL LINE SHOWED H2O2-RESISTANT PROPERTIES, INCREASING THE EXPRESSION OF THE ANTI-OXIDANT GENES CATALASE (CAT), SUPEROXIDE DISMUTASE-1 (SOD1), SUPEROXIDE DISMUTASE-2 (SOD2), AND SUPEROXIDE DISMUTASE-3 (SOD3) AND THE ENZYME ACTIVITIES OF CAT AND INTRACELLULAR SODS. FURTHERMORE, THE RESISTANT CELLS SHOWED INCREASED EXPRESSION LEVELS OF AN EPIGENETICS-RELATED GENE, DNA METHYLTRANSFERASE-1 (DNMT1), WHEN COMPARED TO THE PARENTAL CELLS. INTERESTINGLY, THE OX-MMNK1-L CELL LINE HAD A SIGNIFICANTLY HIGHER CELL PROLIFERATION RATE THAN THE MMNK1 NORMAL CELL LINE. MOREOVER, OX-MMNK1-L CELLS SHOWED PSEUDOPODIA FORMATION AND THE LOSS OF CELL-TO-CELL ADHESION (MULTI-LAYERS) UNDER ADDITIONAL OXIDATIVE STRESS (100 MUM H2O2). THESE FINDINGS SUGGEST THAT H2O2-RESISTANT CELLS CAN BE USED AS A MODEL OF OXIDATIVE STRESS-RELATED CHOLANGIOCARCINOMA GENESIS THROUGH MOLECULAR CHANGES SUCH AS ALTERATION OF GENE EXPRESSION AND EPIGENETIC CHANGES.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
13  1632  32 DNMTS ARE INVOLVED IN TGF-BETA1-INDUCED EPITHELIAL-MESENCHYMAL TRANSITIONS IN AIRWAY EPITHELIAL CELLS. CHRONIC RHINOSINUSITIS (CRS) PATHOGENESIS IS CLOSELY RELATED TO TISSUE REMODELING, INCLUDING EPITHELIAL-MESENCHYMAL TRANSITION (EMT). EPIGENETIC MECHANISMS PLAY KEY ROLES IN EMT. DNA METHYLATION, MEDIATED BY DNA METHYLTRANSFERASES (DNMTS), IS AN EPIGENETIC MARKER THAT IS CRITICAL TO EMT. THE GOAL OF THIS STUDY WAS TO DETERMINE WHETHER DNMTS WERE INVOLVED IN TGF-BETA1-INDUCED EMT AND ELUCIDATE THE UNDERLYING MECHANISMS IN NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. GLOBAL DNA METHYLATION AND DNMT ACTIVITY WERE QUANTIFIED. DNMT EXPRESSION WAS MEASURED USING REAL-TIME PCR (QRT-PCR) IN HUMAN CRS TISSUES. MRNA AND PROTEIN LEVELS OF DNMTS, E-CADHERIN, VIMENTIN, ALPHA-SMA, AND FIBRONECTIN WERE DETERMINED USING RT-PCR AND WESTERN BLOTTING, RESPECTIVELY. DNMT1, DNMT3A, AND DNMT3B GENE EXPRESSION WERE KNOCKED DOWN USING SIRNA TRANSFECTION. MAPK PHOSPHORYLATION AND EMT-RELATED TRANSCRIPTION FACTOR LEVELS WERE DETERMINED USING WESTERN BLOTTING. SIGNALING PATHWAYS WERE ANALYZED USING SPECIFIC INHIBITORS OF MAPK. WE DEMONSTRATED THESE DATA IN PRIMARY NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. GLOBAL DNA METHYLATION, DNMT ACTIVITY, AND DNMT EXPRESSION INCREASED IN CRS TISSUES. DNMT EXPRESSION WAS POSITIVELY CORRELATED WITH LUND-MCKAY CT SCORES. TGF-BETA1 DOSE-DEPENDENTLY INDUCED DNMT EXPRESSION. FURTHER, 5-AZA INHIBITED TGF-BETA1-INDUCED DNMT, SNAIL, AND SLUG EXPRESSION RELATED TO EMT, AS WELL AS P38 AND JNK PHOSPHORYLATION IN A549 CELLS AND TGF-BETA1-INDUCED DNMT EXPRESSION AND EMT IN PRIMARY NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. TGF-BETA1-INDUCED DNMT EXPRESSION LEADS TO DNA METHYLATION AND EMT VIA P38, JNK, SNAIL, AND SLUG SIGNALING PATHWAYS. INHIBITION OF DNMT SUPPRESSED THE EMT PROCESS AND THEREFORE IS POTENTIALLY A CRS THERAPEUTIC STRATEGY.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
14  4303  37 MICRORNA-223 INHIBITS TISSUE FACTOR EXPRESSION IN VASCULAR ENDOTHELIAL CELLS. OBJECTIVE: ATHEROSCLEROSIS IS A CHRONIC INFLAMMATORY PROCESS, IN WHICH VASCULAR ENDOTHELIAL CELLS (ECS) BECOME DYSFUNCTIONAL OWING TO THE EFFECTS OF CHEMICAL SUBSTANCES, SUCH AS INFLAMMATORY FACTOR AND GROWTH FACTORS. TISSUE FACTOR (TF) EXPRESSION IS INDUCED BY THE ABOVE CHEMICAL SUBSTANCES IN ACTIVATED ECS. TF INITIATES THROMBOSIS ON DISRUPTED ATHEROSCLEROTIC PLAQUES WHICH PLAYS AN ESSENTIAL ROLE DURING THE ONSET OF ACUTE CORONARY SYNDROMES (ACS). INCREASING EVIDENCES SUGGEST THE IMPORTANT ROLE OF MICRORNAS AS EPIGENETIC REGULATORS OF ATHEROSCLEROTIC DISEASE. THE AIM OF OUR STUDY IS TO IDENTIFY IF MICRORNA-223 (MIR-223) TARGETS TF IN ECS. METHODS AND RESULTS: BIOINFORMATIC ANALYSIS SHOWED THAT TF IS A TARGET CANDIDATE OF MIR-223. WESTERN BLOTTING ANALYSIS REVEALED THAT TUMOR NECROSIS FACTOR ALPHA (TNF-ALPHA) INCREASED TF EXPRESSION IN AORTA OF C57BL/6J MICE AND CULTURED ECS (EA.HY926 CELLS AND HUVEC) AFTER 4 H TREATMENT. IN TNF-ALPHA TREATED ECS, TF MRNA WAS ALSO INCREASED MEASURED BY REAL-TIME PCR. REAL-TIME PCR RESULTS SHOWED THAT MIR-223 LEVELS WERE DOWNREGULATED IN TNF-ALPHA-TREATED AORTA OF C57BL/6J MICE AND CULTURED ECS. TRANSFECTION OF ECS WITH MIR-223 MIMIC OR MIR-223 INHIBITOR MODIFIED TF EXPRESSION BOTH IN MRNA AND PROTEIN LEVELS. LUCIFERASE ASSAYS CONFIRMED THAT MIR-223 SUPPRESSED TF EXPRESSION BY BINDING TO THE SEQUENCE OF TF 3'-UNTRANSLATED REGIONS (3'UTR). TF PROCOAGULANT ACTIVITY WAS INHIBITED BY OVEREXPRESSING MIR-223 WITH OR WITHOUT TNF-ALPHA STIMULATION. CONCLUSIONS: MIR-223-MEDIATED SUPPRESSION OF TF EXPRESSION PROVIDES A NOVEL MOLECULAR MECHANISM FOR THE REGULATION OF COAGULATION CASCADE, AND SUGGESTS A CLUE AGAINST THROMBOGENESIS DURING THE PROCESS OF ATHEROSCLEROTIC PLAQUE RUPTURE.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
15  4899  26 OXIDATIVE STRESS MEDIATES THE APOPTOSIS AND EPIGENETIC MODIFICATION OF THE BCL-2 PROMOTER VIA DNMT1 IN A CIGARETTE SMOKE-INDUCED EMPHYSEMA MODEL. BACKGROUND: EMPHYSEMA IS A CRUCIAL PATHOLOGICAL CHARACTERISTIC OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD). OXIDATIVE STRESS, APOPTOSIS AND EPIGENETIC MECHANISMS CONTRIBUTE TO THE PATHOGENESIS OF EMPHYSEMA. HOWEVER, AN ATTEMPT TO ACCURATELY IDENTIFY WHETHER THESE MECHANISMS INTERACT WITH EACH OTHER AND HOW THEY ARE TRIGGERED HAS NEVER BEEN CONDUCTED. METHOD: THE TOTAL REACTIVE OXYGEN SPECIES (ROS) LEVEL, PULMONARY APOPTOSIS AND B-CELL LYMPHOMA/LEUKEMIA-2 (BCL-2) EXPRESSION, AN APOPTOSIS REGULATOR, WERE DETECTED IN SAMPLES FROM COPD PATIENTS. BISULFITE SEQUENCING PCR (BSP) WAS CONDUCTED TO OBSERVE THE ALTERATIONS IN THE METHYLATION OF THE BCL-2 PROMOTER IN SPECIMENS. THE DYSREGULATION OF DNA METHYLTRANSFERASE ENZYME 1 (DNMT1), A VITAL DNA METHYLTRANSFERASE ENZYME, IN THE LUNGS OF PATIENTS WAS CONFIRMED THROUGH WESTERN BLOTTING. TO FIND OUT INTERACTIONS BETWEEN OXIDATIVE STRESS AND DNA METHYLATION IN EMPHYSEMA, MOUSE MODELS WERE BUILT WITH ANTIOXIDANT TREATMENT AND DNMT1 SILENCING, AND WERE EXAMINED WITH THE PULMONARY APOPTOSIS, BCL-2 AND DNMT1 LEVELS, AND EPIGENETIC ALTERATIONS OF BCL-2. RESULTS: HIGHER ROS LEVELS AND PULMONARY APOPTOSIS WERE OBSERVED IN COPD PATIENTS THAN IN HEALTHY CONTROLS. DOWNREGULATED BCL-2 EXPRESSION WITH INCREASED PROMOTER METHYLATION AND DNMT1 PROTEIN EXPRESSION WAS FOUND IN COPD PATIENTS. ANTIOXIDANT TREATMENT REDUCED THE LEVEL OF ROS, DNMT1 PROTEIN AND EMPHYSEMATOUS PROGRESSION IN THE SMOKING MODELS. FOLLOWING DNMT1 BLOCKADE, SMOKING MODELS SHOWED IMPROVED LUNG FUNCTION, PULMONARY APOPTOSIS, EMPHYSEMATOUS PROGRESSION, AND INCREASED BCL-2 PROTEIN LEVEL WITH LESS PROMOTER METHYLATION THAN EMPHYSEMA MICE. CONCLUSION: CIGARETTE-INDUCED OXIDATIVE STRESS MEDIATES PULMONARY APOPTOSIS AND HYPERMETHYLATION OF THE BCL-2 PROMOTER IN EMPHYSEMA MODELS THROUGH DNMT1.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
16  3426  31 HUMAN UMBILICAL ARTERY ENDOTHELIAL CELLS FROM LARGE-FOR-GESTATIONAL-AGE NEWBORN HAVE INCREASED ANTIOXIDANT EFFICIENCY AND GENE EXPRESSION. OBESITY IS A PUBLIC HEALTH PROBLEM WORLDWIDE, AND ESPECIALLY IN WOMEN IN REPRODUCTIVE AGE WHERE MORE THAN ONE IN THREE HAVE OBESITY. MATERNAL OBESITY IS ASSOCIATED WITH AN INCREASED MATERNAL, PLACENTAL, AND NEWBORN OXIDATIVE STRESS, WHICH HAS BEEN PROPOSED AS A CENTRAL FACTOR IN VASCULAR DYSFUNCTION IN LARGE-FOR-GESTATIONAL-AGE (LGA) NEWBORN. HOWEVER, CELLULAR AND MOLECULAR MECHANISMS BEHIND THIS EFFECT HAVE NOT BEEN ELUCIDATED. UNTREATED HUMAN UMBILICAL ARTERY ENDOTHELIAL CELLS (HUAEC) FROM LGA (LGA-HUAEC) PRESENTED HIGHER O(2)(-) LEVELS, SUPEROXIDE DISMUTASE ACTIVITY AND HEME OXYGENASE 1 MESSENGER RNA (MRNA) LEVELS, PARALLELED BY REDUCED GSH:GSSG RATIO AND NRF2 MRNA LEVELS. IN RESPONSE TO AN OXIDATIVE CHALLENGE (HYDROGEN PEROXIDE), ONLY HUAEC FROM LGA EXHIBITED AN ENHANCED GLUTATHIONE PEROXIDASE 1 (GPX1) EXPRESSION, AS WELL AS A MORE EFFICIENT ANTIOXIDANT MACHINERY MEASURED BY THE BIOSENSOR PROBE, HYPER. AN OPEN STATE OF CHROMATIN IN THE TSS REGION OF GPX1 IN LGA-HUAEC WAS EVIDENCED BY THE DNASE-HS ASSAY. ALTOGETHER, OUR DATA INDICATE THAT LGA-HUAEC HAVE AN ALTERED CELLULAR AND MOLECULAR ANTIOXIDANT SYSTEM. WE PROPOSE THAT A CHRONIC PRO-OXIDANT INTRAUTERINE MILIEU, AS EVIDENCED IN PREGESTATIONAL OBESITY, COULD INDUCE A MORE EFFICIENT ANTIOXIDANT SYSTEM IN FETAL VASCULAR CELLS, WHICH COULD BE MAINTAINED BY EPIGENETIC MECHANISM DURING POSTNATAL LIFE.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
17  5860  32 SULFORAPHANE PREVENTS ANGIOTENSIN II-INDUCED CARDIOMYOPATHY BY ACTIVATION OF NRF2 THROUGH EPIGENETIC MODIFICATION. NUCLEAR FACTOR ERYTHROID 2-RELATED FACTOR (NRF2) IS AN IMPORTANT REGULATOR OF CELLULAR ANTIOXIDANT DEFENCE. WE PREVIOUSLY SHOWED THAT SFN PREVENTED ANG II-INDUCED CARDIAC DAMAGE VIA ACTIVATION OF NRF2. HOWEVER, THE UNDERLYING MECHANISM OF SFN'S PERSISTENT CARDIAC PROTECTION REMAINS UNCLEAR. THIS STUDY AIMED TO EXPLORE THE POTENTIAL OF SFN IN ACTIVATING CARDIAC NRF2 THROUGH EPIGENETIC MECHANISMS. WILD-TYPE MICE WERE INJECTED SUBCUTANEOUSLY WITH ANG II, WITH OR WITHOUT SFN. ADMINISTRATION OF CHRONIC ANG II-INDUCED CARDIAC INFLAMMATORY FACTOR EXPRESSION, OXIDATIVE DAMAGE, FIBROSIS AND CARDIAC REMODELLING AND DYSFUNCTION, ALL OF WHICH WERE EFFECTIVELY IMPROVED BY SFN TREATMENT, COUPLED WITH AN UP-REGULATION OF NRF2 AND DOWNSTREAM GENES. BISULFITE GENOME SEQUENCING AND CHROMATIN IMMUNOPRECIPITATION (CHIP) WERE PERFORMED TO DETECT THE METHYLATION LEVEL OF THE FIRST 15 CPGS AND HISTONE H3 ACETYLATION (AC-H3) STATUS IN THE NRF2 PROMOTER REGION, RESPECTIVELY. THE RESULTS SHOWED THAT SFN REDUCED ANG II-INDUCED CPG HYPERMETHYLATION AND PROMOTED AC-H3 ACCUMULATION IN THE NRF2 PROMOTER REGION, ACCOMPANIED BY THE INHIBITION OF GLOBAL DNMT AND HDAC ACTIVITY, AND A DECREASED PROTEIN EXPRESSION OF KEY DNMT AND HDAC ENZYMES. TAKEN TOGETHER, SFN EXERTS ITS CARDIOPROTECTIVE EFFECT THROUGH EPIGENETIC MODIFICATION OF NRF2, WHICH MAY PARTIALLY CONTRIBUTE TO LONG-TERM ACTIVATION OF CARDIAC NRF2.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
18  3449  35 HYPERMETHYLATION OF THE NRF2 PROMOTER INDUCES FERROPTOSIS BY INHIBITING THE NRF2-GPX4 AXIS IN COPD. BACKGROUND: NUCLEAR FACTOR E2-RELATED FACTOR 2 (NRF2) IS INVOLVED IN OXIDATIVE STRESS AND LUNG INFLAMMATION AND REGULATES THE ETIOLOGY OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD). FERROPTOSIS IS CHARACTERIZED BY THE ACCUMULATION OF LIPID REACTIVE OXYGEN SPECIES (ROS) VIA FERROUS ION-DEPENDENT FENTON REACTIONS AND IS INVOLVED IN COPD. HOWEVER, THE ROLE OF NRF2 IN FERROPTOSIS AND ITS EPIGENETIC REGULATION IN THE PATHOGENESIS OF COPD REMAIN UNCLEAR. METHODS: FERROPTOSIS WAS DETECTED BY 4-HNE, MDA, C11BODIPY, DCFH-DA, PEALS' STAINING AND CCK-8 ASSAYS. QPCR AND WESTERN BLOTTING WERE PERFORMED TO EXAMINE THE NRF2 LEVELS IN PERIPHERAL LUNG TISSUES, PRIMARY EPITHELIAL CELLS COLLECTED FROM PATIENTS WITH COPD AND SUBJECTS WITH NORMAL PULMONARY FUNCTION (NEVER-SMOKER [CONTROL-NS]; SMOKER [CONTROL-S]), AND CIGARETTE SMOKE EXTRACT (CSE)-TREATED HUMAN BRONCHIAL EPITHELIAL (HBE) CELLS. ELISA WAS USED TO QUANTIFY IL-8 AND IL-1BETA LEVELS. METHYLATION OF THE NRF2 PROMOTER WAS ANALYZED BY BISULFITE SEQUENCING AND PYROSEQUENCING. RESULTS: FERROPTOSIS WAS INVOLVED IN COPD AND GLUTATHIONE PEROXIDASE 4 (GPX4) EXPRESSION WAS DOWNREGULATED IN THE COPD GROUP. REACTIVE OXYGEN SPECIES (ROS), LIPID PEROXIDES AND MDA WERE INCREASED, BUT GPX4 AND SOD WERE EXHAUSTED IN CSE-TREATED HBE CELLS. THE PRODUCTION OF IL-1BETA AND IL-8 WAS PROMOTED IN HBE CELLS IN RESPONSE TO CSE BUT COULD BE REVERSED BY THE FERROPTOSIS INHIBITOR FER-1. THE NRF2 LEVEL WAS SIGNIFICANTLY DECREASED IN THE COPD GROUP COMPARED WITH THE CONTROL-S AND CONTROL-NS GROUPS. INCREASED NRF2 EXPRESSION ENHANCED GPX4 AND SOD LEVELS AND INHIBITED FERROPTOSIS AND PROINFLAMMATORY CYTOKINES IN THE SUPERNATANT. INHIBITION OF GPX4 REVERSED THE EFFECT OF NRF2 OVEREXPRESSION AND PROMOTED FERROPTOSIS. TWO SPECIFIC CPG SITES WITHIN THE NRF2 PROMOTER WERE HYPERMETHYLATED IN THE COPD GROUP. SIMILARLY, CSE-TREATED HBE CELLS EXHIBITED HYPERMETHYLATION OF THE NRF2 GENE. CONCLUSION: NRF2 EXPRESSION WAS DOWNREGULATED IN THE LUNGS OF COPD PATIENTS DUE TO HYPERMETHYLATION OF THE NRF2 PROMOTER, INHIBITING NRF2/GPX4 AND FERROPTOSIS, WHICH IS RELATED TO THE INITIATION AND PROGRESSION OF COPD. TARGETING NRF2/GPX4 MAY INHIBIT FERROPTOSIS, WHICH COULD PROVIDE STRATEGIES TO DELAY OR TREAT COPD.	2021	
                                                                                                                                                                                                                                                                                                                                                          
19  3036  33 GENISTEIN AMELIORATES RENAL FIBROSIS THROUGH REGULATION SNAIL VIA M6A RNA DEMETHYLASE ALKBH5. RENAL TUBULE-INTERSTITIAL FIBROSIS IS RELATED TO CHRONIC KIDNEY DISEASE PROGRESSION AND A TYPICAL FEATURE OF THE AGING KIDNEY. EPIGENETIC MODIFICATIONS OF FIBROSIS-PRONE GENES REGULATE THE DEVELOPMENT OF RENAL FIBROSIS. AS A KIND OF "EPIGENETIC DIET", SOY ISOFLAVONE GENISTEIN WAS REPORTED TO HAVE RENAL PROTECTIVE ACTION AND EPIGENETIC-MODULATING EFFECTS. HOWEVER, ITS RENAL PROTECTION ROLE AND UNDERLYING MECHANISMS ARE YET TO BE FULLY CLARIFIED. HEREIN, WE SHOWED THAT GENISTEIN EXHIBITS A DEMONSTRABLE ANTI-FIBROTIC EFFECT ON KIDNEY IN VIVO UUO (UNILATERAL URETERAL OCCLUSION) MODEL AND RENAL EPITHELIAL CELLS IN VITRO MODEL. THE MECHANISM IS STRONGLY ASSOCIATED WITH EPITHELIAL-TO-MESENCHYMAL TRANSITION AND M6A RNA DEMETHYLASE ALKBH5. MOUSE FIBROTIC KIDNEYS INDUCED BY UUO EXHIBITED ADVERSE EXPRESSION OF RENAL FIBROSIS-RELATED PROTEINS AND SIGNIFICANT INCREASES IN THE TOTAL M6A LEVEL. AS AN ERASER, ALKBH5 SHOWED SEVERER SUPPRESSION IN THE RENAL FIBROSIS PROCESS. HOWEVER, GENISTEIN PRETREATMENT RESTORED ALKBH5 LOSS REMARKABLY AND REDUCED RENAL FIBROSIS, ABNORMAL PROTEIN, AND INFLAMMATORY MARKERS. THE EXAMINATION OF POSSIBLE MECHANISMS REVEALED THAT GENISTEIN PROMOTED ALKBH5 AND MAYBE INDUCED THE LEVEL OF MRNA M6A METHYLATION IN SOME EPITHELIAL-TO-MESENCHYMAL TRANSITION-RELATED TRANSCRIPTION FACTORS. WE FOUND SNAIL WAS THE CRITICAL REGULATOR AND CRITICAL FOR THE PROTECTIVE ROLE OF GENISTEIN. TO VERIFY THE RELATIONSHIP BETWEEN ALKBH5 AND SNAIL, WE GENERATED KNOCKDOWN AND OVEREXPRESSION OF ALKBH5 CELLS IN VITRO. ALKBH5 KNOCKDOWN ENHANCED THE MESENCHYMAL PHENOTYPE MARKER ALPHA-SMOOTH MUSCLE ACTIN AND SNAIL EXPRESSION. IN AGREEMENT, OVEREXPRESSION ALKBH5 INCREASED EPITHELIAL ADHESION MOLECULE E-CADHERIN AND REDUCED SNAIL EXPRESSION. IN CONCLUSION, GENISTEIN INCREASED RENAL ALKBH5 EXPRESSION IN UUO-INDUCED RENAL FIBROSIS AND REDUCED RNA M6A LEVELS AND AMELIORATES RENAL DAMAGES.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
20  5850  41 SUBEROYLANILIDE HYDROXAMIC ACID (SAHA) REDUCES FIBROSIS MARKERS AND DEACTIVATES HUMAN STELLATE CELLS VIA THE EPITHELIAL-MESENCHYMAL TRANSITION (EMT). HEPATIC FIBROSIS IS KNOWN AS THE ACCUMULATION OF CONNECTIVE TISSUE SECONDARY TO CHRONIC DAMAGE TO THE LIVER. EPITHELIAL-MESENCHYMAL TRANSITION (EMT) CORRESPONDING INCREASE IN LIVER FIBROGENESIS WAS SHOWN WITH IMMUNOHISTOCHEMISTRY AND PCR-BASED STUDIES. SUBEROYLANILIDE HYDROXAMIC ACID (SAHA), A SYNTHETIC COMPOUND APPROVED AS A HISTONE DEACETYLASE INHIBITOR (HDAC) BY THE FDA TO TREAT CUTANEOUS T-CELL LYMPHOMA IS UNDER INVESTIGATION FOR THE TREATMENT OF LUNG AND RENAL FIBROSIS. EXPERIMENTAL MODELING FOR HEPATIC FIBROSIS CAN BE CONSTRUCTED WITH AN LX2 CELL LINE ISOLATED FROM HUMAN HEPATIC STELLATE CELLS (HSCS). IN THIS STUDY, WE AIMED TO INVESTIGATE THE MODULATION OF SAHA IN THE PATHOGENESIS OF LIVER FIBROSIS BY DETECTING THE LEVELS OF PROTEINS; (E-CADHERIN (E-CAD), N-CADHERIN (N-CAD), VIMENTIN (VIM), AND GENES; E-CAD, N-CAD, VIM, TRANSFORMING GROWTH FACTOR-BETA (TGF-BETA), ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), TYPE 1 COLLAGEN (COL1A1), TYPE 3 COLLAGEN (COL3A1)) THAT PLAY A SIGNIFICANT ROLE IN EMT WITH THE LX2 CELL LINE. WE ALSO EVALUATED THE ACTION OF SAHA WITH CELL PROLIFERATION, CLONOGENIC, AND MIGRATION ASSAY. CELL PROLIFERATION WAS PERFORMED BY FLOW CYTOMETRY. ALL THE PROTEIN LEVELS WERE DETERMINED BY WESTERN BLOT ANALYSIS, AND GENE EXPRESSION LEVELS WERE MEASURED BY REAL-TIME PCR. OUR STUDY OBSERVED THAT SAHA TREATMENT DECREASED CELL VIABILITY, COLONY FORMATION AND MIGRATION IN LX2 CELLS. WE FOUND THAT SAHA INCREASED E-CAD EXPRESSION LEVEL, WHILE IT DECREASED N-CAD, VIM, COL1A1, COL3A1, ALPHA-SMA TGF-BETA GENES EXPRESSION LEVELS. SAHA DECREASED THE LEVEL OF E-CAD, N-CAD, AND VIM PROTEIN LEVELS. WE THOUGHT THAT SAHA POSSESSES POTENT ANTIFIBROTIC AND ANTI-EMT PROPERTIES IN LX2.	2021