Anti-oxidative and anti-apoptotic effects and molecular mechanisms of catalpol against H_2O_2-induced oxidative damage in pancreatic β cells (INS-1 cells).
10.19540/j.cnki.cjcmm.20220119.402
- Author:
Xin XIAO
1
;
Wen-Hua XU
1
;
Xiao-Qing ZHANG
2
;
Jun-Feng DING
1
;
Yue JIANG
2
;
Jun TU
2
Author Information
1. Research Center for Differentiation and Development of Chinese Medicine Basic Theory & Jiangxi Province Key Laboratory of Chinese Medicine Etiopathogenisis, Jiangxi University of Chinese Medicine Nanchang 330004, China.
2. Research Center for Differentiation and Development of Chinese Medicine Basic Theory & Jiangxi Province Key Laboratory of Chinese Medicine Etiopathogenisis, Jiangxi University of Chinese Medicine Nanchang 330004, China Key Laboratory of Traditional Chinese Medicine Pharmacology of Jiangxi Province Nanchang 330004, China.
- Publication Type:Journal Article
- Keywords:
INS-1 cells;
apoptosis;
catalpol;
oxidative damage;
β cell function
- MeSH:
Apoptosis;
Glucose/metabolism*;
Insulin/metabolism*;
Insulin-Secreting Cells/metabolism*;
Iridoid Glucosides;
Kelch-Like ECH-Associated Protein 1/metabolism*;
NF-E2-Related Factor 2/metabolism*;
Oxidative Stress;
Reactive Oxygen Species/metabolism*;
Superoxide Dismutase/metabolism*
- From:
China Journal of Chinese Materia Medica
2022;47(16):4403-4410
- CountryChina
- Language:Chinese
-
Abstract:
The present study investigated the anti-oxidative and anti-apoptotic effects and molecular mechanisms of catalpol on the H_2O_2-induced pancreatic β-cells(INS-1 cells).The oxidative damage model of INS-1 cells was induced and optimized by the stimulation of H_2O_2 of different concentrations for different time.CCK-8 assay was used to detect cell viability after catalpol intervention(1, 5, 10, 20, 40, 80, and 160 μmol·L~(-1)) for 24 h.Intracellular reactive oxygen species(ROS), superoxide dismutase(SOD), and lipid peroxide malondialdehyde(MDA) were measured by DCFH-DA fluorescent probe, WST-1, and TBA respectively.Moreover, the apo-ptotic effect was detected by AO-EB and Annexin V-FITC/PI staining.In addition, the protein expression levels were detected by Wes-tern blot, and intracellular insulin concentration was measured by ELISA.The results showed that the oxidative damage model of INS-1 cells was stably induced by 50 μmol·L~(-1) H_2O_2 treatment for 2 h, and catalpol at 1-80 μmol·L~(-1) did not affect cell viability of INS-1 cells.Compared with the conditions in the model group, 1, 5, and 10 μmol·L~(-1) catalpol intervention for 2 h could protect INS-1 cells from oxidative damage(P<0.001), reduce ROS and MDA, increase SOD, and inhibit excessive cell apoptosis.Moreover, 1, 5, and 10 μmol·L~(-1) catalpol could also up-regulate the phosphorylation of nuclear transcription factor NF-E2 related factors, negatively regulate Kelch-like ECH-associated protein 1(Keap1), phosphorylation of extracellular signal-regulated kinase(ERK), and heme oxyge-nase 1(HO-1), and promote the protein expression of pancreatic-duodenal homeobox factor-1(PDX-1) and glucose transporter 2(GLUT2).In addition, 1, 5, and 10 μmol·L~(-1) catalpol increased insulin secretion of INS-1 cells under oxidative damage in the high-glucose culture medium, indicating function recovery of pancreatic β cells.PDX-1 is a key nuclear transcription factor of pancreatic β cell function that directly regulates GLUT2 and insulin synthesis, and affects glucose homeostasis.In conclusion, catalpol can reduce the oxidative damage and apoptosis of INS-1 cells, activate antioxidant pathway, protect the function of pancreatic β cells, and improve insulin synthesis and secretion.
