Effects of MD2 gene silencing on high glucose-induced proliferation inhibition, apoptosis and inflammation in rat cardiomyocytes.
10.12047/j.cjap.5834.2019.057
- Author:
Zhong-Min LIN
1
;
Guo-Rong CHEN
1
;
Quan-Bo ZHANG
1
;
Fang WANG
1
;
Lan-Ting XIANG
1
;
Qiong-Jie CAO
2
Author Information
1. Department of Pathology, the First Affiliated Hospital, Wenzhou Medical University, Wenzhou 325000, China.
2. State Key Laboratory of Ophthalmology, Optometry and Vision Science, Eye Hospital of Wenzhou Medical University, Wenzhou 325000, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Apoptosis;
Cell Proliferation;
Cells, Cultured;
Cytokines;
metabolism;
Gene Silencing;
Glucose;
Inflammation;
JNK Mitogen-Activated Protein Kinases;
metabolism;
Lymphocyte Antigen 96;
genetics;
Myocytes, Cardiac;
cytology;
Rats;
p38 Mitogen-Activated Protein Kinases;
metabolism
- From:
Chinese Journal of Applied Physiology
2019;35(3):273-278
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effects of myeloid differentiation-2 (MD2) gene silencing on high glucose-induced proliferation inhibition, apoptosis and inflammation in rat cardiomyocytes.
METHODS:The immortalized rat cardiomyocyte cell line H9C2 were transfected with MD2 small interfering RNA (si-MD2) and negative control for 24 h, then stimulated with high glucose (HG) for 48 h. RT-qPCR was performed to detect the mRNA levels of MD2 and inflammatory factors TNF-α, IL-1β and IL-6. MTS and flow cytometry were used to evaluate cell proliferation, cell cycle and apoptosis rate. Western blot was used to detect protein expression levels and phosphorylation levels.
RESULTS:The mRNA and protein levels of MD2 in H9C2 cells were dramatically decreased after transfected with si-MD2 (P＜0.01). After stimulation of high glucose, the mRNA levels of inflammatory factors, the cells in G0/G1 phase , the cell apoptosis rate and the protein level of cleaved Caspase-3 were significantly increased, while the cell proliferation ability was decreased (P＜0.01). MD2 gene silencing antagonized the effects of high glucose on cell proliferation, cell cycle, cell apoptosis and the mRNA levels of TNF-α, IL-1β , IL-6(P＜0.05). Western blot analysis showed that the phosphorylation levels of extracellular signal-regulated kinase(ERK1/2), P38 mitogen-activated protein kinase(P38 MAPK) and C-Jun N-terminal kinase(JNK) protein were increased significantly in H9C2 cells treated with high glucose, which could be reversed by silencing of MD2 (P＜0.01).
CONCLUSION:This study demonstrates that MD2 gene silencing reverses high glucose-induced myocardial inflammation, apoptosis and proliferation inhibition via the mechanisms involving suppression of ERK, P38 MAPK, JNK signaling pathway.
