Journal of the Korean Pediatric Society 2001;44(11):1262-1268.
Published online November 15, 2001.
The Effect of Tumor Necrosis Factor-α in Cultured Neonatal Rat Cardiomyocytes
Su Jin Kim1, Dong Seok Lee1, Ok Ko2, Il Soo Mun2, Bok Hyun Ko2, Yong Wook Jung2
1Department of Pediatrics, College of Medicine, Dongguk University, Kyungju, Korea
2Department of Anatomy, College of Medicine, Dongguk University, Kyungju, Korea
배양된 신생 백서 심근 세포에 대한Tumor Necrosis Factor-α의 효과
김수진1, 이동석1, 고옥2, 문일수2, 고복현2, 정용옥2
1동국대학교 의과대학 소아과학교실
2동국대학교 의과대학 해부학교실
Correspondence: 
Dong Seok Lee, Email: lds117@dumc.or.kr
Abstract
Purpose
: Tumor necrosis factor-α(TNF-α) is a pro-inflammatory cytokine that has been implicated in the pathogenesis of cardiovascular disease. Serum levels of TNF-α are elevated in many human cardiac related pathogenic conditions, including heart failure. It is well known that TNF-α inhibits myocardial contractility and induces apoptosis of adult rat cardiomyocytes via stimulation of TNF receptor 1. But pathophysiologically relevant low levels of TNF-α can not induce apoptosis of neonatal cardiomyocytes. So, we evaluated the effects of different concentrations of TNF-α 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-α 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-α did not induce apoptosis compared with controls(10.5±3.5% : 10.4±4.3%). And high dose TNF-α 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-α treatment. LDH levels after TNF-α treatment was not significant compared with control(control : low : high, 3.2±0.1% : 3.1±0.2% : 3.3±0.3%).
Conclusion
: Our results suggest that high concentration of TNF-α alone can not induce apoptosis and significant cytotoxicity in neonatal rat cardiomyocytes.
Key Words: TNF-α, Neonatal rat, Cardiomyocyte, Apoptosis


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