• Toxicological Research
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• 제13권1_2호
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• pp.61-69
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• 1997

Exposure of splenocytes to 2-chloroethylethyl sulfide (CEES) resulted in the cell death, and the cytotoxicity of CEES was prevented by inhibitors of lysosomal hydrolases. Therefore, it has been postulated that the cytotoxicity of CEES may be partially due to the lysosomal labilization. This study, based on this mechanism, was undertaken to determine whether aminoglycoside antibiotics as inhibitors of lysosomal phospholipases and their combinations with other lysosome stabilizers can be useful as a treatment to reduce the CEES toxicity in mice. 2-Chloroethylethyl sulfide (20 mg/kg body weight) was injected ip into female ICR mice, and candidate compounds were administered ip before or after the CEES challenge. Kanamycin (40 mg/kg body weight) as effective as deferoxamine (100 mg/kg body weight) enhanced the survival rate after 5 days of intoxication from 10% of control to 50 - 60%. The most effective was found to be the combination of kanamycin, cycloheximide, deferoxamine and dextrose showing an almost full protection against 2LD50 of CEES. Consistent with the protection of the CEES toxicity, the decrease of body weight in mice intoxicated with CEES was effectively prevented by kanamycin or its combinations. It is suggested that kanamycin or its combination (kanamycin, cycloheximide, deferoxamine and dextrose) would be one of effective antidotes against the CEES poisoning in mice.

• 대한화장품학회지
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• 제45권4호
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• pp.389-397
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• 2019

리소좀(lysosome)은 진핵세포에서 에너지 대사 및 세포 내 소화 작용에 관여하는 세포 소기관으로 protease, nuclease, glycosidase, lipase, phosphatase 들이 다수 존재한다. 우리는 선행 연구결과들을 통해 난백 리소좀의 멜라닌 색소 탈색능을 보고하였다[8]. 그러나 B16F10 melanocyte 세포주에서 난백 리소좀에 의한 멜라닌 함량 변화 및 피부장벽 조절 연구는 거의 보고되지 않았다. 따라서 우리는 계란 난백(egg white)으로부터 추출한 lysosome-related organelle extract (LOE)에 의한 세포 내 멜라닌 함량 변화 및 피부장벽 강화 효과를 규명하고자 하였다. 먼저 LOE의 미백 효능을 확인하기 위해 B16F10 세포주를 이용하여 세포독성 평가를 진행하였다. B16F10 세포주에서 LOE에 의한 세포독성은 0에서 20 mg/mL 농도에서 관찰되지 않았으나, 40 mg/mL 부터 세포독성이 관찰되어 이후 모든 실험에서 최대 농도값을 20 mg/mL로 설정하였다. 먼저 LOE를 이용한 melanin contents assay 결과, 음성 대조군인 α-MSH 처리군 대비 LOE 처리군 5, 10, 20 mg/mL 농도에서 61.5 ± 4.0%, 61.4 ± 7.3%, 58.3 ± 8.3%로 세포 내 멜라닌 함량이 감소되는 것을 확인하였고, 20 mg/mL 농도 조건에서 MITF 발현 억제도 관찰하였다. LOE의 피부 장벽에 미치는 영향을 관찰하기 위해 각질형성세포주(HaCaT)를 이용하여 TEER (trans-epithelial electrical resistance) assay를 수행한 결과, LOE에 의해 농도 의존적으로 TEER 저항값이 증가하여 LOE가 피부장벽 강화에도 효과가 있음을 알 수 있었다. 또한 피부 염증 유발을 위한 TNF-α 처리조건에서도 LOE는 TEER 저항값을 증가시켜 염증 유발 조건에서도 LOE에 의해 피부장벽이 정상적으로 회복되었음을 알 수 있었다. 마지막으로 cell migration assay를 통해 LOE에 의한 세포이동 촉진 효과를 관찰한 결과, LOE는 세포분열 및 세포이동을 촉진시켰다. 위 결과들을 통해 LOE는 미백 기능 뿐 아니라 피부재생 및 피부장벽 강화에도 효과를 나타내는 소재이며, 효소안정화 및 제형화 기술이 접목된다면 향후 새로운 미백 기능성 화장품 소재로도 개발될 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.718-722
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• 2015

Techniques for immobilizing effective enzymes on nanoparticles for stabilization of the activity of free enzymes have been developing as a pharmaceutical field. In this study, we examined the effect of three different pH conditions of phosphate buffer, as a dissolving solvent for lysosomal enzymes, on the direct immobilization of lysosomal enzymes extracted from Hen's egg white and Saccharomyces cerevisiae. Titanium(IV) oxide (TiO₂) nanoparticles, which are extensively used in many research fields, were used in this study. The lysosomal enzymes immobilized on TiO₂ under each pH condition were evaluated to maintain the specific activity of lysosomal enzymes, so that we can determine the degree of melanin treatment in lysosomal enzymes immobilized on TiO₂. We found that the immobilization efficiency and melanin treatment activity in both lysosomal enzymes extracted from Hen's egg white and S. cerevisiae were the highest in an acidic condition of phosphate buffer (pH 4). However, the immobilization efficiency and melanin treatment activity were inversely proportional to the increase in pH under alkaline conditions. In addition, enhanced immobilization efficiency was shown in TiO₂ pretreated with a divalent, positively charged ion, Ca²⁺, and the melanin treatment activity of immobilized lysosomal enzymes on TiO₂ pretreated with Ca²⁺ was also increased. Therefore, this result suggests that the immobilization efficiency and melanin treatment activity of lysosomal enzymes can be enhanced according to the pH conditions of the dissolving solvent.