High glucose induces H9 c2 cardiomyocyte hypertrophy through Ca2+-CaN-NFAT3 signaling pathway / 中国病理生理杂志

ABSTRACT

AIM:To study the morphological changes of cardiac H9c2 cells during the developmental process of fetal rat.METHODS:Embryonic rat heart-derived H9c2 cells were maintained in DMEM supplemented with 10%fetal bovine serum.The H9c2 cells were plated at a density of 6 000 cells/cm and divided into 5 groupsH9c2 cells were trea-ted with 5 mmol/L glucose, 25 mmol/L glucose, 50 mmol/L glucose, Norvasc (25 nmol/L) +25 mmol/L glucose, or Norvasc (25 nmol/L)+50 mmol/L glucose for 48 h.The morphology of H9c2 cells was observed.The cell surface area was measured by Image-Pro Plus 6.1 software.Fluorescence spectrophotometry was used to detect the concentration of in-tracellular calcium ion ( [ Ca2+] i ) in the cardiomyocytes.The concentration of CaN in the cell was measured by ELISA. The mRNA expression of CaNAβ, NFAT3 and β-MHC in the cells was detected by real-time PCR.The protein levels of CaNAβ, NFAT3 and β-MHC in cultural H9c2 cells were detected by Western blot.RESULTS: The mean area of the cells, the mean fluorescence value of [ Ca2+] i and the concentration of CaN in 25 mmol/L glucose group were higher than those in 5 mmol/L glucose group, and those were lower than those in 50 mmol/L glucose group.After treated with Nor-vasc, those results decreased significantly.The expression of CaNAβ, NFAT3 andβ-MHC at mRNA and protein levels in 25 mmol/L glucose group was higher than those in 5 mmol/L glucose group, but was lower than those in 50 mmol/L glu-cose group .The expression of CaNAβ, NFAT3 andβ-MHC at mRNA and protein levels decreased significantly in Norvasc treatment group.CONCLUSION:Ca2+-CaN-NFAT3 signaling pathway is perhaps involved in high glucose-induced H9c2 cardiomyocyte hypertrophy.