ABSTRACT
OBJECTIVE@#To investigate the effects of hydrogen sulfide (HS) on the negatively regulation of cardiomyocyte hypertrophy and the relationship between the effect of HS with miRNA-133a-mediated Ca/calcineurin/NFATc4 signal pathway.@*METHODS@#Cardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (Leica). The expression of brain natriuretic peptide(BNP), β-myosin heavy chain(β-MHC), cystathionase (CSE), miRNA-133a, calcineurin (CaN) were detected by qRT-PCR. The protein expressions of CaN、nuclear factors of activated T cells (NFATc4) were detected by Western blot. The concentration of HS in the cardiomyocyte was detected by Elisa. The concentration of intracellular calcium was measured by calcium imaging using confocal microscope. The nuclear translocation of NFATc4 was checked by immuno-fluorescence cell staining technique.@*RESULTS@#①The level of system of CSE/HS and expression of miRNA-133a were significantly reduced in cardiomyocyte hypertrophy. Pretreatment with NaHS increased the concentration of HS and the expression of miRNA-133a mRNA in cardiomyocytes, and suppressed cardiomyocyte hypertrophy. ②The concentration of intracellular calcium, the expression of CaN and nulear protein NFATc4 were significantly increased, and the nuclear translocation of NFATc4 were obviously enhanced in cardiomyocyte hypertrophy. NaHS pretreatment markedly inhibited these effects of ISO induced cardiomyocyte hypertrophy. ③Application of antagomir-133a reversed the inhibitory effects of NaHS on cardiomyocyte hypertrophy, and increased the influx of intracellular calcium, and elevated the expression of CaN and nuclear protein NFATc4, and enhanced the nuclear translocation of NFATc4.@*CONCLUSIONS@#HS can negatively regulate cardiomyocyte hypertrophy. The effects might be associated with HS increasing expression of miRNA-133a and inhibiting inactivation of Ca/calcineurin/NFATc4 signal pathway.
Subject(s)
Animals , Rats , Calcineurin , Metabolism , Cardiomegaly , Metabolism , Cells, Cultured , Cystathionine gamma-Lyase , Metabolism , Hydrogen Sulfide , Metabolism , MicroRNAs , Metabolism , Myocytes, Cardiac , Metabolism , Myosin Heavy Chains , Metabolism , NFATC Transcription Factors , Metabolism , Natriuretic Peptide, Brain , Metabolism , Nerve Tissue Proteins , Metabolism , Signal TransductionABSTRACT
AIM:To investigate the effects of ursolic acid ( UA) on the migration and invasion of human lung cancer cell line A549, and to explore its mechanism .METHODS:The cell viability was detected by MTT assay .The ex-pression of miRNA-133a was detected in the A549 cells treated with UA by real-time PCR.The miRNA-133a mimics and inhibitor were transfected into the A 549 cells, and the transfection efficiency was analyzed by real-time PCR.The cell mi-gratory and invasive abilities were determined by wound healing and Transwell methods , respectively .RESULTS:The via-bility of the human lung cancer A549 cells was significantly inhibited by UA in a dose-dependent manner (P<0.05).IC50 of UA (24 h) for lung cancer A549 cells was 31.04 μmol/L.UA treatment significantly inhibited the migratory and inva-sive abilities of A549 cells in a concentration-dependent manner , accompanied by significantly elevation of miRNA-133a expression.The mimics and inhibitor of miRNA-133a significantly upregulated and downregulated the expression of miRNA -133a in the transfected A549 cells, respectively.In addition, the viability of the A549 cells was decreased extremely after tansfected with the miRNA-133a mimics (P<0.01), so did the results of the cell migration and invasion test .The A549 cells tansfected with the miRNA-133a inhibitor showed an opposite changes of the cell viability , migration and invasion . CONCLUSION:UA inhibited the viability , migration and invasion of lung cancer A 549 cells by elevating the expression of miRNA-133a.
ABSTRACT
tonomous Prefecture,Enshi Hubei 445000,China;2.Graduate School,Guangdong Medical College,Zhanjiang Guangdong 524001,China) Objective To study the relationship of miRNA-133a-3p and hypoxia in rat cardiomyocytes,and to detect the expression of the probable target genes.Methods SD rats cardiomyocytes were cultivated,and treated in hypoxia condition(37 ℃,5%CO2 ,1%O2 )for 0 hour,4 hours,8 hours,12 hours,16 hours,20 hours and 24 hours.The miRNA-133a-3p level was detected by qRT-PCR,and its biological information and target genes were predicted by targetScan picTar and miRanda,etc.Then we carried out qRT-PCR and Western blot to detect the expression of the probable target genes.Results miRNA-133a-3p expression was much higher in 4 hours hypoxia (P =0.000),then its expression gradually declined,and stabilized after 12 hours hypoxia.Bioinformatics prediction found that TGF-β1 may be its target.The mR-NA expression of TGF-β1 was obviously higher in 4 hours hypoxia than those in 0 hour and 24 hours(P Objective To study the relationship of miRNA-133a-3p and hypoxia in rat cardiomyocytes,and to detect the expression of the probable target genes.Methods SD rats cardiomyocytes were cultivated,and treated in hypoxia condition(37 ℃,5%CO2 ,1%O2 )for 0 hour,4 hours,8 hours,12 hours,16 hours,20 hours and 24 hours.The miRNA-133a-3p level was detected by qRT-PCR,and its biological information and target genes were predicted by targetScan picTar and miRanda,etc.Then we carried out qRT-PCR and Western blot to detect the expression of the probable target genes.Results miRNA-133a-3p expression was much higher in 4 hours hypoxia (P =0.000),then its expression gradually declined,and stabilized after 12 hours hypoxia.Bioinformatics prediction found that TGF-β1 may be its target.The mR-NA expression of TGF-β1 was obviously higher in 4 hours hypoxia than those in 0 hour and 24 hours(P <0.05).And the protein expression of TGF-β1 was significantly higher in 4 hours hypoxia(P <0.05).Conclusion miRNA-133a-3p may participate in cardiomyocytes necrosis in rats through regulating TGF-β1.