The Effect of Tumor Necrosis Factor-alpha in Cultured Neonatal Rat Cardiomyocytes.
- Author:
Su Jin KIM
1
;
Dong Seok LEE
;
Ok KO
;
Il Soo MUN
;
Bok Hyun KO
;
Yong Wook JUNG
Author Information
1. Department of Pediatrics, College of Medicine, Dongguk University, Kyungju, Korea. 1ds117@dumc.or.kr
- Publication Type:Original Article
- Keywords:
TNF-alpha;
Neonatal rat;
Cardiomyocyte;
Apoptosis
- MeSH:
Adult;
Animals;
Apoptosis;
Cardiovascular Diseases;
Cell Culture Techniques;
Cell Survival;
Collagenases;
Heart Failure;
Humans;
Lactic Acid;
Muscle Cells;
Myocytes, Cardiac*;
Necrosis;
Rats*;
Receptors, Tumor Necrosis Factor;
Tumor Necrosis Factor-alpha*
- From:Journal of the Korean Pediatric Society
2001;44(11):1262-1268
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: Tumor necrosis factor-alpha(TNF-alpha) is a pro-inflammatory cytokine that has been implicated in the pathogenesis of cardiovascular disease. Serum levels of TNF-alpha are elevated in many human cardiac related pathogenic conditions, including heart failure. It is well known that TNF-alpha inhibits myocardial contractility and induces apoptosis of adult rat cardiomyocytes via stimulation of TNF receptor 1. But pathophysiologically relevant low levels of TNF-alpha can not induce apoptosis of neonatal cardiomyocytes. So, we evaluated the effects of different concentrations of TNF-alpha in cultured rat neonatal cardiomyocytes : apoptosis or necrosis. METHODS: Neonatal ventricular myocytes were isolated from 3-day-old rats by stepwise collagenase dissociation, and the cells were cultured for 3 days. After that, cardiomyocytes were treated with low(25 ng/mL) and high(250 ng/mL) concentration of TNF-alpha for 48 hours. Apoptosis was determined by terminal deoxynucleotidyl transfer-mediated end labelling(TUNEL) staining, and cell viability was evaluated by lactate dehydrogenase(LDH) measurements using cell culture supernatants. RESULTS: Low dose TNF-alpha did not induce apoptosis compared with controls(10.5 +/- 3.5% : 10.4 +/- 4.3%). And high dose TNF-alpha also did not induce significant apoptosis(10.2 +/- 3.6% : 10.4 +/- 4.3%). There was no detectable morphological changes of cardiomyocytes after low and high concentration of TNF-alpha treatment. LDH levels after TNF-alpha treatment was not significant compared with control(control : low : high, 3.2 +/- 0.1% : 3.1 +/- 0.2% : 3.3 +/- 0.3%). CONCLUSIONS: Our results suggest that high concentration of TNF-alpha alone can not induce apoptosis and significant cytotoxicity in neonatal rat cardiomyocytes.
