Inhibition Of High-glucose-induced Inflammation By Pioglitazone In Neonatal Cardiac Myocytes Of Rat

ObjectivesThe rats myocardial cell stimulated by high glucose was used to induce themyocardial cell injury. Giving pioglitazone treatment to observe the influence ofinflammatory markers of myocardial cells, and p-p38MAPK and NF-κB proteinexpression changes.Methods1. Cultured cardiomyocytes were randomly divided into4groups:1) normal group:cardiomyocytes were cultured in DMEM containing5.5mmol/L glucose as a normalcontrol.2) hight glucose group: cardiomyocytes were cultured in high glucoseDMEM containing30mmol/L glucose.3) low dose group: cardiomyocytes werecultured in high glucose DMEM containing30mmol/L glucose and10μmol/Lpioglitazone.4) high dose group: cardiomyocytes were cultured in high glucoseDMEM containing30mmol/L glucose and20μmol/L pioglitazone. After additional48hours culture, cells and medium were collected for further assays.2. The content of LDH and Inflammation factors in culture medium were detected byLDH KIT and ELISA KIT respectively. The protein expressions of p-p38MAPK wasmeasured by Western Blot. Myocardial cell nucleoprotein was extracted by Nuclearand Cytoplasmic Protein Extraction Kit, then, the cell nucleoprotein expressions ofp65was measured by Western Blot.Results1. The content of LDH in culture mediumResults show that compared with normal control group, the content of LDH of the high glucose group was significantly increased (p<0.01), this suggests that highglucose can cause myocardial cell inflammatory damage. With pioglitazone treatment,the content of LDH was significantly lower (p<0.05) suggest that Pioglitazone hasprotective effect on myocardial cell.2. The content of inflammatory cytokines IL-6and TNF-αResults show that compared with normal control group, the content of highglucose group inflammation factor of IL-6, TNF-α were significantly increased(p<0.05; p<0.01). This suggests that high glucose can cause myocardial cellinflammatory damage. With pioglitazone treatment (low dose group and high dosegroup), inflammatory factors of IL-6, TNF-α levels were significantly lower (p<0.05;p<0.05) suggest that pioglitazone treatment can significantly reduce inflammatoryinjury of cells.3. The activity of transcription factors the NF-κBResults show that compared with normal group, high glucose group ofmyocardial cell nuclei p65protein expression was significantly increased (p<0.01),while giving pioglitazone (low dose and high dose) myocardial cell nucleus p65protein expression was significantly reduced (p<0.05). Results suggest that givingthe pioglitazone significantly revrese the increased of expression of nucleus p65caused by high glucose.4. p38MAPK activationResults show that compared with normal group, p-p38MAPK protein expressionof the high glucose group increased significantly (p<0.01), and treated withpioglitazone (low dose group and high dose group) p-p38MAPK elevated proteinexpression was improved obviously (p<0.05; p<0.01).Conclusions1. High glucose cultivation of myocardial cells can induced cell damage, along withthe production of inflammatory cytokines, the NF-κB activity increased andp-p38MAPK protein expression increased. 2. Pioglitazone’s treatment to high glucose culture of rat myocardial cells can relieveinflammatory cell damage, transcription factor NF-κB activity was alleviated andprotein expression of p-p38MAPK was decreased. Pioglitazone can inhibitp-p38MAPK protein expression and the NF-κB activity.