Diabetes Macroangiopathy
Glucagon-like peptide-1 ameliorates ferroptosis induced by advanced glycosylation end products in human aortic endothelial cells
Hu Kexin, Tang Longjia, Zhang Hui, Ye Xiao, Wo Enkang, Xiang Tingting, He Yongyu, Ma Shumin, Wu Xiaohong
Published 2023-05-20
Cite as Chin J Diabetes Mellitus, 2023, 15(5): 409-415. DOI: 10.3760/cma.j.cn115791-20221130-00687
Abstract
ObjectiveTo investigate the effect of glucagon-like peptide-1 (GLP-1) on advanced glycation end products (AGE)-induced ferroptosis in human aortic vascular endothelial cells (HAEC) and its possible mechanism.
MethodsHAEC was divided into 6 groups: control group, AGE group, ferrostatin-1 (Fer-1) group, AGE+GLP-1 (7-37) group, AGE+GLP-1 (9-36) group and AGE+GLP-1 (28-36) group. Each group has three parallel groups. After 48 h of culture, cell viability and malondialdehyde (MDA) content were detected by the kit, mRNA expression levels of ferroptosis related genes [prostaglandin-endoperoxide synthase 2 (PTGS2), acyl-CoA synthetase long-chain family member 4 (ACSL4), glutathione peroxidase 4 (GPX4)] were detected by real-time fluorescence quantitative polymerase linked reaction, and the protein levels of GPX4, ACSL4, liver kinase B1 (LKB1), phosphorylated LKB1 (p-LKB1), adenosine monophosphate-activated protein kinase (AMPK) and phosphorylated AMPK (p‐AMPK) were detected by Western blotting. One-way analysis of variance (ANOVA) and t-test were used for comparison between groups.
ResultsCompared with the control group, mitochondrial morphology was abnormal in the AGE group, and cell vitality was significantly inhibited (P<0.05),the cell viability of the AGE+Fer-1 group was significantly increased (P<0.05). In the AGE group, the mRNA level of the positive regulator of ferroptosis gene PTGS2 was up-regulated (P<0.05), the mRNA level of ACSL4 was increased (P<0.05), and the mRNA level of the negative regulator gene GPX4 was down-regulated (P<0.05). The expression of above genes was reversed in AGE+Fer-1 group (P<0.05). Compared to the AGE group, the viability of HAEC cells in GLP-1 and its small molecule fragment group was significantly increased (P<0.05), the content of MDA in cells was decreased, and the degree of lipid peroxidation was decreased (P<0.05). The expression of GPX4 was up-regulated (P<0.05), and the expression of ACSL4 was down-regulated (P<0.05). The expression of p-LKB1 and p-AMPK were significantly increased (P<0.05).
ConclusionGLP-1 can reduce ferroptosis induced by AGE in HAEC, and the mechanism may be related to the activation of LKB1/AMPK signaling pathway.
Key words:
Glycosylation end products, advanced; Glucagon-like peptide-1; Human aortic endothelial cells; Ferroptosis
Contributor Information
Hu Kexin
Department of Endocrinology, Zhejiang Provincial People′s Hospital, Affiliated People′s Hospital, Hangzhou Medical College, Hangzhou 310014, China
Tang Longjia
Department of Endocrinology, Zhejiang Provincial People′s Hospital, Affiliated People′s Hospital, Hangzhou Medical College, Hangzhou 310014, China
Zhang Hui
Lab of Public Platform, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Ye Xiao
Department of Endocrinology, Zhejiang Provincial People′s Hospital, Affiliated People′s Hospital, Hangzhou Medical College, Hangzhou 310014, China
Wo Enkang
School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou 310013, China
Xiang Tingting
School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou 310013, China
He Yongyu
School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou 310013, China
Ma Shumin
School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou 310013, China
Wu Xiaohong
Department of Endocrinology, Zhejiang Provincial People′s Hospital, Affiliated People′s Hospital, Hangzhou Medical College, Hangzhou 310014, China
