사염화에틸렌에 의한 병아리배자의 심장기형 및 기능변화에 관한 연구

• 영문명: A Study for Cardiovascular Malformations and Functional Changes Induced by Tetrachloroethylene in Developing Chick Embryos
• 발행기관: 대한해부학회
• 저자명: 김 완(Wan Kim) 김원규(Won-Kyu Kim) 최태열(Tae-Yeal Choi) 조자연(Ja-Yeon Cho)
• 간행물 정보: 『대한해부학회지』제32권 제2호, 181~189쪽, 전체 9쪽
• 주제분류: 의약학 > 의학일반
• 파일형태: PDF
• 발행일자: 1999.04.30

국문 초록

본 연구는 사염화에틸렌 (tetrachloroethylene)을 병아리 배자에 투여하여 심혈관기형과의 유관성을 조사하였다. 또한 투여 후 초기 변화로써 세포자사와 연관된 세포손상 및 혈류량 감소와 같은 생리기능적 장애등을 조사하여, 심혈관기형의 형태발생학적 기전에의 접근을 시도하고자 하였다. Hamburger-Hamilton 발생 27기 및 28기의 수정된 계란을 90~96시간 배양하여 실험군에는 40 μM, 50 μM의 두가지 농도의 사염화에틸렌을 mineral oil 0.05 ml에 용해시켜 융모요막에 주입하였고, 대조군은 식염수, mineral oil군으로 나누어 각각 0.05 ml의 식염수와 mineral oil을 투여하였다. 사염화에틸렌 주입 후 6분에서 30분간 심전도 변화를 추적하였고, pulse doppler velocity meter로 배대동맥 (dorsal aorta)을 통해 혈류의 속도를 측정하였으며, 실험군과 대조군의 14일 배자들을 고정하고 조직표본을 제작한 후 광학현미경으로 관찰하였고, 전자현미경조직표본을 제작하여 투과전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 40~50 μM 농도에서 심혈관 기형이 가장 잘 유발되어, 여러 가지 형태의 심장기형이 실험군에서 통계적으로 유의하게 (p<0.01) 관찰되었고, 상당한 치사율을 동반하였다. 심장기형으로는 대형심실사이막결손, 대형심방사이막결손, 심장속막융기 결손, 양대혈관 오른심실기시 (double outlet right ventricle)와 동맥줄기개존을 동반한 단심실이었으며, 이중 대형심실사이막결손이 실험군 410예중 48예 (11.7%)로 가장 많았다. 심전도상에선 투여 6분에서 30분 후에 심실세동을 포함한 부정맥이 보였고, 도플러 (doppler) 측정상 심전도 이상과 일치하여 배대동맥의 혈류량 감소가 두드러졌다. 광학현미경 소견상 개별 세포사를 주 소견으로 하는 대량의 세포괴사를 보였으며, 전자현미경적소견상 배자심근 (embryonic myocardium)의 세포손상을 나타내는 소견이 관찰되었다.

영문 초록

Chlorinated solvents are recognized as well-known drinking water contaminants and potent teratogens for developing embryos. Among theses compounds, tetrachloroethylene was studied to evaluate the teratogenic effects in detail. This study was also undertaken to understand the teratogenetic mechanisms by assessing the relationship between the physiological impairments and cellular degeneration including apoptosis induced by tetrachloroethylene, and the cardiovascular malformations. Fertilized white leghorn eggs (n=530) were incubated for 90~96 hours up to Hamburger-Hamilton stage 27~28. To the experimental group, tetrachloroethylene was dissolved in mineral oil at concentrations of 40 μM and 50 μM and was injected into the chorioallantoic membrane. One half of the control was injected with saline and the other half with mineral oil. The experimental animals are studied for ECG changes, and light and electron microscopic observation. Two silver thread electrodes were used to record the ECG. ECG changes were recorded for 6~30 minutes after tetrachloroethylene injections. In parallel studies, blood velocity through the dorsal aorta was measured by a pulse Doppler velocity meter with 20 MHz probe. The specimens were stained with hematoxylin and eosin for light microscopic study and the Hitachi H-800 EM was used for transmission electron microscopic evaluation. The embryos underwent formalin fixation on day 14, and the presence of malformations was observed by a microscopy. The results obtained are as follows: Repeated injections of 40~50 μM tetrachloroethylene appeared to have a strong influence on the formation of cardiovascular malformations. Various types of cardiac malformations were observed in the experimental group compared to control group, that were statistically significant (p<0.01) and many embryos also died. The malformations observed were large ventricular septal defect, endocardial cushion defect, double outlet right ventricle and truncus arteriosus associated with single ventricle. Large ventricular septal defect was most common type of cardiovascular abnormalities. Arrythymias including ventricular fibrillations were evident on ECG 6~30 minutes after the injections. Blood flow through the dorsal aorta was markedly decreased in concordance with the ECG changes. A large amount of individual cell death suggesting apoptosis was seen on light microscopic evaluation and the electron microscope revealed cellular degeneration of the embryonic myocardium. We concluded that tetrachloroethylene, like other chloride compounds has potent teratogenic effects. The cardiac malformations were probably caused by decreased blood flow due to physiofunctional changes and cellular necrosis associated with apoptosis. By including birds and mammals in further experiments, we hope to elucidate an embryological clue into how early cardiovascular malformations are caused.

키워드

사염화에틸렌 병아리배자 선천성심장기형 심장기능변화 Tetrachloroethylene Developing Chick Embryos Cardiovascular Malformations and Functional Changes