[Glucagon like peptide-1 inhibits high glucose-induced injury of oxidative stress in cardiomyocytes of neonatal rats.] [Article in Chinese]
BIAN Yun-Fei, WANG Dong-Xue, YANG Hui-Yu, XIAO Chuan-Shi*
Department of Cardiology, the Second Affiliated Hospital of Shanxi Medical University, Taiyuan 030001, China
Abstract
The present study was to investigate the effect of glucagon like peptide-1 (GLP-1) on high glucose-induced oxidative stress of cardiomyocytes and the possible role of the PI3K-Akt signal path in this process in the neonatal SD rats. With enzymatic digestion and immunofluorescence identification, cardiomyocytes after 72–96 h of primary culture were used in experiment. The cells were divided into 5 groups: normal control group, high glucose group, high glucose + GLP-1 group, high glucose + GLP-1 + LY294002 group and high osmolarity control group. The content of MDA was detected by TBA colouration method. The content of SOD was detected by xanthine oxidase method. The change of NADPH P47phox subunit mRNA quantity was detected by PCR gel electrophoresis. The level of ROS was detected by flow cytometry, and was also observed by fluorescence microscope. The DNA ladder was examined by agarose gel electrophoresis, and the cell apoptosis was determined by Annexin-V-FITC/PI flow cytometry, and the phosphorylation of Akt was determined by Western blotting. Compared with those in the normal control group, in the high glucose group, the cells grew poorly, and the beating rate was significantly lower (P < 0.05); The apoptotic rate was significantly increased (P < 0.05); The MDA content was increased (P < 0.05); It showed the typical DNA ladder, which is the characteristic of apoptosis; The SOD activity was decreased (P < 0.05); The level of intracellular ROS increased (P < 0.05); And the expression of NADPH P47phox subunit mRNA was increased; However the phosphorylation level of Akt was decreased. Pretreatment with GLP-1 improved the above-mentioned parameters and decreased the expression of NADPH P47phox subunit mRNA (P < 0.05). However, compared with the high glucose + GLP-1 group, LY294002, an inhibitor of PI3K-Akt signal path, attenuated the protective effect of GLP-1 in the high glucose + GLP-1 + LY294002 group. It is suggested that GLP-1 plays a protective role in the high glucose-induced injury and apoptosis of cardiomyocytes, and the PI3K-Akt signal path is involved in this process.
Key words: neonatal rat; cardiomyocyte; high glucose; glucagon-like peptide-1; PI3K-Akt signal path; LY294002
Received: 2011-02-18 Accepted: 2011-05-30
Corresponding author: 肖传实 E-mail: ganxibaozhongxin@sina.com
Citing This Article:
BIAN Yun-Fei, WANG Dong-Xue, YANG Hui-Yu, XIAO Chuan-Shi. [Glucagon like peptide-1 inhibits high glucose-induced injury of oxidative stress in cardiomyocytes of neonatal rats.] [Article in Chinese]. Acta Physiol Sin 2011; 63 (4): 387-395 (in Chinese with English abstract).