Effect of ryanodine receptor 2 gene silencing on ischemia-reperfusion injury of rat myocardial cells.
- Author:
Zhu-ying GUO
1
;
Qiang JIAO
;
Shi-ting WANG
;
Mang-hua XU
;
Feng-hou GAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Apoptosis; drug effects; genetics; Cells, Cultured; Gene Silencing; immunology; physiology; Membrane Potential, Mitochondrial; drug effects; immunology; Myocardial Reperfusion Injury; immunology; pathology; Myocytes, Cardiac; drug effects; pathology; Oxygen; metabolism; RNA Interference; RNA, Small Interfering; pharmacology; Rats; Rats, Sprague-Dawley; Reperfusion Injury; immunology; pathology; Ryanodine Receptor Calcium Release Channel; drug effects; genetics
- From: Chinese Journal of Pathology 2008;37(11):760-764
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVESTo block the synthesis of ryanodine receptor 2 (RyR2) in myocardial cells by RNA interference and to investigate its biological impact on ischemia-reperfusion (I/R) in rat myocardial cells.
METHODSRat myocardial cells were isolated and cultured for an I/R model in vitro. RNA interference technique was used to block the synthesis of RyR2 in myocardial cells. Changes of LDH level, apoptosis, RyR2 mRNA expression and cytosolic Ca(2+) concentration were analyzed accordingly.
RESULTSMyocardial cells after I/R manipolation were severely injuried (LDH leakage, 125 IU/L vs 12 IU/L, P < 0.05), apoptosis (60.1% vs 5.5%, P < 0.05), significant cytosolic Ca(2+) overload (21.2 vs 7.6, P < 0.05) and remarkable mitochondrial membrane potential loss (37.2 vs 85.1, P < 0.05). However, no visible change of RyR2 was observed (20.1 vs 22.7, P > 0.05). Pre-treatment with RyR2 specified siRNA demonstrated suppressed expression of RyR2 (6.8 vs 20.1, P < 0.05), increased mitochondrial membrane potential (55.8 vs 37.2, P < 0.05), attenuated cytosolic Ca(2+) overload (8.6 vs 21.2) and cellular apoptosis (31.2% vs 60.1%, P < 0.05).
CONCLUSIONRyR2 gene silencing enables to protect myocardial cells from I/R injury in vitro.
