김지욱
(Department of Chemistry and Biology, Korea Science Academy of KAIST)
,
공희정
(Biotechnology Research Division, National Institute of Fisheries Research)
,
김영환
(Applied Acoustics Laboratory, Korea Science Academy of KAIST)
,
강광일
(Department of Chemistry and Biology, Korea Science Academy of KAIST)
초음파는 세포사멸을 포함하여 의학 및 생물학분야에 널리 응용되고 있으나 그 정확한 기작에 대해선 논쟁의 여지가 있다. 본 연구에서는 40 kH 초음파 조사시스템을 단세포 효모에 적합하게 개발하고 세포사멸 유도시 40 kH 초음파의 생물학적 현상을 살펴보았다. 아이오딘화 칼륨 선량 측정법을 이용하여 1.5 ml 실험튜브에 40 kH 초음파 조사 시스템의 최적 조건을 맞추어 세포사멸을 시간 의존적 방식으로 연구하였고 초음파 조사과정동안 온열효과와는 별개로 세포 사멸이 관찰되었다. 40 kH 초음파와 과산화수소의 동시 처리는 세포사멸에 상조적인 효과가 관찰되어 활성산소가 40 kH 초음파사멸에 관련이 있었다. 그러나 활성산소 저해제, NAC(N-acetyl-Lcysteine)는 초음파에 의한 세포사멸에 약한 영향만을 미쳤고 다른 세포사멸, 괴사억제제[글리실리진(glycyrrhizin) 또는 zVAD-fmk] 역시도 세포사멸을 완전히 억제하진 못하였다. 본 연구를 통하여 40 kH 초음파에 의한 세포사멸에는 온열효과나 활성산소만으로 사멸이 유도되지는 않는 것으로 보인다.
초음파는 세포사멸을 포함하여 의학 및 생물학분야에 널리 응용되고 있으나 그 정확한 기작에 대해선 논쟁의 여지가 있다. 본 연구에서는 40 kH 초음파 조사시스템을 단세포 효모에 적합하게 개발하고 세포사멸 유도시 40 kH 초음파의 생물학적 현상을 살펴보았다. 아이오딘화 칼륨 선량 측정법을 이용하여 1.5 ml 실험튜브에 40 kH 초음파 조사 시스템의 최적 조건을 맞추어 세포사멸을 시간 의존적 방식으로 연구하였고 초음파 조사과정동안 온열효과와는 별개로 세포 사멸이 관찰되었다. 40 kH 초음파와 과산화수소의 동시 처리는 세포사멸에 상조적인 효과가 관찰되어 활성산소가 40 kH 초음파사멸에 관련이 있었다. 그러나 활성산소 저해제, NAC(N-acetyl-Lcysteine)는 초음파에 의한 세포사멸에 약한 영향만을 미쳤고 다른 세포사멸, 괴사억제제[글리실리진(glycyrrhizin) 또는 zVAD-fmk] 역시도 세포사멸을 완전히 억제하진 못하였다. 본 연구를 통하여 40 kH 초음파에 의한 세포사멸에는 온열효과나 활성산소만으로 사멸이 유도되지는 않는 것으로 보인다.
Ultrasound has been widely used for biological and medical applications including induction of cell death, but a precise mechanism of induced cell death by ultrasound is controversial. In this study, an irradiation system with 40 kHz ultrasound was developed for a suitable cell death test of a repre...
Ultrasound has been widely used for biological and medical applications including induction of cell death, but a precise mechanism of induced cell death by ultrasound is controversial. In this study, an irradiation system with 40 kHz ultrasound was developed for a suitable cell death test of a representative unicellular organism, yeast, and used to study the biological effect of ultrasound on inducing cell death. Potassium Iodide (KI) dosimetry was used to devise an optimal system that successfully delivers 40 kHz ultrasound and produces reactive oxygen species in a 1.5 ml Eppendorf tube. Cell death was observed in an ultrasound transmission time-dependent fashion in this system. Thermal effect during irradiation was not observable in ultrasound induced cell death. Co-treatment of 40 kHz ultrasound and hydrogen peroxide showed a synergistic effect in inducing cell death. This finding suggests that 40 kHz ultrasound is related to reactive oxygen species formation. However, NAC (N-acetyl-L-cysteine) oxygen scavenger slightly inhibited the cell death by 40 kHz ultrasound. It was also found that 40 kHz ultrasound induced cell death was slightly inhibited by inhibitors of necrosis or apoptosis (glycyrrhizin or zVAD-fmk). This study suggests that cell death induced by 40 kHz ultrasound may not be exclusively related to reactive oxygen species formation and thermal effects in irradiated yeast cells.
Ultrasound has been widely used for biological and medical applications including induction of cell death, but a precise mechanism of induced cell death by ultrasound is controversial. In this study, an irradiation system with 40 kHz ultrasound was developed for a suitable cell death test of a representative unicellular organism, yeast, and used to study the biological effect of ultrasound on inducing cell death. Potassium Iodide (KI) dosimetry was used to devise an optimal system that successfully delivers 40 kHz ultrasound and produces reactive oxygen species in a 1.5 ml Eppendorf tube. Cell death was observed in an ultrasound transmission time-dependent fashion in this system. Thermal effect during irradiation was not observable in ultrasound induced cell death. Co-treatment of 40 kHz ultrasound and hydrogen peroxide showed a synergistic effect in inducing cell death. This finding suggests that 40 kHz ultrasound is related to reactive oxygen species formation. However, NAC (N-acetyl-L-cysteine) oxygen scavenger slightly inhibited the cell death by 40 kHz ultrasound. It was also found that 40 kHz ultrasound induced cell death was slightly inhibited by inhibitors of necrosis or apoptosis (glycyrrhizin or zVAD-fmk). This study suggests that cell death induced by 40 kHz ultrasound may not be exclusively related to reactive oxygen species formation and thermal effects in irradiated yeast cells.
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제안 방법
An irradiation system with ultrasound at 40 kHz frequency was developed (Fig. 1) and used to test the KI ionization for optimal position. Fig.
In this study, a system that transmits 40 kHz ultrasound was developed and optimized to study yeast as a representative unicellular organism. Using an ultrasound transmission system, yeast cell death was examined.
The yeast cells are easy to manipulate and have many advantages including applicability to human cancer cells. Then, we examined the effect of 40 kHz ultrasound on the cell death and analyzed the thermal effect, radicals effects and other possible effects. We used the 50 W output of ultrasound generator and had worked in condition of standing wave.
대상 데이터
The yeast Saccharomyces cerevisiae BY4743 was purchased from EUROSCARF (Germany). The obtained results are applicable to human cells.
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