Protective effects of adenoviral-mediated sphingosine kinase gene on ischemia reperfusion injury of heart
- VernacularTitle:腺病毒介导的鞘氨醇激酶1高表达对心脏缺血再灌注损伤的保护作用观察
- Author:
Jin ZHANG
;
Hong WANG
;
Ying LU
- Publication Type:Journal Article
- Keywords:
sphingosine kinases;
gene therapy;
myocardial reperfusion injury
- From:
Medical Journal of Chinese People's Liberation Army
1981;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective Sphingosine kinase 1 (SPK1) has been identified as a central mediator of ischemia preconditioning, and it has been shown to protect reactive oxygen species (ROS)-induced cardiomyocytes death. The present study aims at investigating the protective effects of adenovirus-mediated sphingosine kinase 1 gene (Ad-SPK1) transfer on ischemia-reperfusion induced cardiac injury. Methods Wistar rats were anesthetized and a left thoracotomy was performed. About 5?108 PFU of Ad-SPK1 in total volume of 100?l was injected intramyocardially into four separate sites of the left ventricular wall with a 30-gauge needle. The control rats received the same injection of adenovirus carrying green fluorescent protein gene (Ad-GFP). Three days later, hearts were isolated and subjected to ischemia/reperfusion (I/R) (30min/30min) ex vivo. Heart performance was evaluated by measuring the left ventricular systolic pressure (LVSP) and left ventricular end-diastolic pressure (LVEDP). The incidence of arrhythmia was recorded. Cardiomyocyte viability was detected by the detection of the release of creatine kinase (CK). The cardiac SPK1 activity was measured using an enzyme method. The forced and the total SKP1 expression was analyzed by immunoblotting with anti-FLAG and anti-SPK1 antibodies. Results The cardiac SPK1 activity was increased by about five-fold by injection of Ad-SPK1, compared to Ad-GFP control group. A more potent performance and a lower incidence of arrhythmia were observed in Ad-SPK1-injected hearts during the reperfusion period, compared with Ad-GFP-injected ones. Enzymatic activity assay showed that creatine kinase release was also less in Ad-SPK1-injected hearts. Conclusion Adenovirus-mediated SPK1 gene transfer efficiently protects heart from injury induced by ischemia-reperfusion.
