Effects of high glucose induced primary cardiomyocytes injury on necroptosis and the related mechanism.
10.12047/j.cjap.5710.2019.035
- Author:
Ting Ting FANG
1
;
Rui Ping CAO
1
;
Hong Wei YE
1
;
Shan Feng MA
1
;
Qin GAO
1
Author Information
1. Department of Physiology, Bengbu Medical College, Bengbu 233030, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cells, Cultured;
Cytokines;
metabolism;
Glucose;
adverse effects;
Humans;
Myocytes, Cardiac;
cytology;
pathology;
Necrosis;
Oxidative Stress;
Reactive Oxygen Species;
metabolism
- From:
Chinese Journal of Applied Physiology
2019;35(2):160-164
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To observe whether necroptosis was happened in high glucose (HG) - induced primary cardiomyocytes injury and to investigate the likely mechanism.
METHODS:The primary cultured cardiomyocytes were divided into 4 groups (n=9): control group (the cardiomyocytes were incubated with 5.5 mmol/L glucose for 48 h), HG group (the cardiomyocytes were incubated with 30 mmol/L glucose for 48 h), HG + necrostatin-1 (Nec-1) group (the cardiomyocytes was co-incubated with necroptosis inhibitor Nec-1 at 100 μmol/L and HG for 48 h) and hypertonic pressure group (HPG, the cardiomyocytes was co-incubated with 5.5 mmol/L glucose and 24.5 mmol/L mannitol for 48 h). Cell viability was measured by MTT method, reactive oxygen species (ROS) generation was measured by DHE staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) were tested by ELISA method. The mRNA and protein expressions of necroptosis related genes receptor interacting serine/threonine protein kinase 1 (RIP1), RIP3, mixed lineage kinase domain-like protein (MLKL) were tested by quantitative real-time PCR and Western blot.
RESULTS:The results showed HG intervention decreased cardiomyocytes viability, increased ROS generation, up-regulated the levels of TNF-α, IL-6 and IL-1β, increased RIP1, RIP3, MLKL expressions at mRNA and protein levels. Nec-1 treatment attenuated HG-induced increased cardiomyocytes viability, reduced ROS generation, down-regulated the levels of TNF-α, IL-6 and IL-1β, decreased RIP1, RIP3, MLKL expressions at mRNA and protein levels.
CONCLUSION:Necroptosis was happened in high glucose-induced primary cardiomyocytes injury. Inhibition of necroptosis can reduce high glucose-induced cardiomyocytes damage, may be related to inhibition of oxidative stress and depression of inflammative factors releasing.
