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The effect and underlying mechanisms of pioglitazone on apoptosis of H9C2 cardiomyocytes
induced by high glucose/palmitic acid
CHEN Xu, SHI Kaijia, WANG Jiao, YAO Shan, ZHOU Tian, JIANG Chenglong, CHEN Xiaopan
2022, 50 (4):
337-342.
doi: 10.11958/20211872
Objective To investigate the effect and underlying mechanisms of pioglitazone (PGZ) on apoptosis of rat
H9C2 cardiomyocytes induced by high glucose (HG) /palmitic acid (PA). Methods H9C2 cells were cultured and
stimulated with different concentrations of PA and HG for 12, 24 and 48 h to determine the best concentration by CCK-8
assays. The cell model of hyperglycemia and hyperlipid injury was established by co-cultivation of 0.1 mmol/L PA and
50 mmol/L HG. The injury cells stimulated with different concentrations of PGZ for 12, 24 and 48 h were determined the
best intervention time and concentrations. H9C2 cells were divided into the five groups: the control group (25 mmol/L
glucose), the solvent group (PA solvent+DMSO), the hyperglycemia and hyperlipid group (HGPA), the H-PGZ group
(10 μmol/L PGZ+HGPA) and the L-PGZ group (5 μmol/L PGZ+HGPA). The apoptosis rate was detected by fluorescent
probe of AnnexinV-FITC/PI. The level of reactive oxidative species (ROS) was detected by DCFH-DA with flow cytometry.
The MDA content and SOD level were assayed by specific kits. The protein expression levels of AKT, P-AKT, Caspase3, C-Caspase3, BCL-2 and BAX were detected by Western blot assay. After 24 h of NAC (1 mmol/L N-acetylcysteine) +HGPA
and HGPA treatment, the above AKT pathway and apoptosis-related protein expression levels were detected again. Results
CCK-8 showed that H9C2 cell viability decreased successively in the PA group after treatment with 0, 0.05, 0.1, 0.2 and
0.4 mmol/L PA for 48 h（P＜0.05). In 12 h, compared with the 25 mmol/L glucose group, the cell proliferation increased in
the 50, 75 and 100 mmol/L HG groups, and the cell proliferation increased successively in 24 h and 48 h in the 25, 35, 50,
75 and 100 mmol/L HG groups. Compared with the control group, the cell viability decreased within 48 hours in the HGPA
group (P＜0.05). Compared with the HGPA group, the cell viability decreased at 12 h in the 80 μmol/L PGZ treatment group
(P＜0.05). The cell viability increased at 24 h in the 10 μmol/L PGZ group, while cell viability decreased in the 40 and
80 μmol/L groups (P＜0.05). At 48 h, the cell viability increased in the 5, 10 and 20 μmol/L groups, while the cell viability
decreased in the 40 and 80 μmol/L groups (P＜0.05). Compared with the control group, the apoptosis rate, ROS level, MDA
content, C-Caspase3 expression and BAX expression increased in the HGPA group, while SOD level, P-AKT expression
and BCL-2 expression decreased (P＜0.05). The apoptosis rate, ROS, MDA, C-caspase3 and Bax decreased in turn in the
HGPA group, the L-PGZ group and the H-PGZ group, while the protein expression levels of SOD, P-AKT, Caspase3 and
BAX increased in turn (P＜0.05). Compared with the HGPA group, C-Caspase3 expression decreased in the NAC+HGPA
group, while P-AKT increased (P＜0.05). Conclusion PGZ has an inhibitory effect on ROS, which can promote activation
of the AKT pathway, and reduce the apoptosis of H9C2 cardiomyocytes induced by HG/PA.
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