Journal of fish pathology (한국어병학회지)

The Korean Society of Fish Pathology (한국어병학회)
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Natural killer cell activity of olive flounder Paralichthys olivaceus following intramuscular injection of toltrazuril derivative N-(4-(4-Fluorophenoxy)-3-methylphenyl) acetamide

톨트라주릴 합성유도체, N-(4-(4-Fluorophenoxy)-3-methylphenyl) acetamide 근육 주사에 따른 넙치의 자연살해세포(Natural killer cell) 활성 검사

  • Sang Hyup Park (Department of Aquatic Life Medicine, Gangneung-Wonju National University) ;
  • Jung Eui Kim (Department of Aquatic Life Medicine, Gangneung-Wonju National University) ;
  • Jeong-wan Do (Pathology Division, National Institute of Fisheries Science) ;
  • Ah Ran Kim (Pathology Division, National Institute of Fisheries Science) ;
  • Yi Kyung Kim (Department of Aquatic Life Medicine, Gangneung-Wonju National University)
  • 박상협 (강릉원주대학교 수산생명의학과) ;
  • 김정의 (강릉원주대학교 수산생명의학과) ;
  • 도정완 (국립수산과학원 병리연구과) ;
  • 김아란 (국립수산과학원 병리연구과) ;
  • 김이경 (강릉원주대학교 수산생명의학과)
  • Received : 2024.05.23
  • Accepted : 2024.06.12
  • Published : 2024.06.30
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Abstract

This study assessed the impact of the toltrazuril derivative N-(4-(4-Fluorophenoxy)-3-methylphenyl) acetamide on natural cytotoxic cell (NCC) activity of olive flounder, Paralichthys olivaceus spleen. Five groups of fifteen olive flounder, comprising non-treatment and vehicle control groups, were randomly assigned. N-(4-(4-Fluorophenoxy)-3-methylphenyl) acetamide was injected intramuscularly at doses of 120, 150 and 200 mg/kg body weight; a total of ten injections were given over the course of 30 days. The NK activity of flounder splenic cells was evaluated against YAC-1, mouse lymphoma cells or HINAE cells with a choice of co-cultivation times of 4 or 18 hrs. In case of YAC-1 co-culture we observed a significant increase in cytotoxicity at a dose of 200 mg/kg, up to 3.06 times more than that of the control group. Only the trial with the 4 hrs co-culture produced a significant difference in the HINAE cell experiment; the experimental group at the 200 mg/kg dose exhibited the maximum cytotoxicity, demonstrating 2.3 times more cytotoxicity than the control group. Furthermore, the expression level of IL-12b was markedly induced in the group with 200 mg/kg, which was 6.62 times greater than that of the control group. In terms of the altered NK cell activity, the repeated high doses of N-(4-(4-Fluorophenoxy)-3-methylphenyl) acetamide can cause changes in the normal performance of immune function.

본 연구에서는 넙치(Paralichthys olivaceus)에서 N-(4-(4-Fluorophenoxy)-3-methylphenyl) acetamide 주사 투여에 따른 넙치 비장의 NK cell 활성을 평가하기 위하여 120, 150, 200 mg/kg 용량으로 설정하고 3일에 1회, 30일 동안 총 10회의 주사를 투여하였다. 표적세포(Target cell)로는 쥐의 임파종 세포인 YAC-1 cell과 넙치의 HINAE cell을 사용하였고, 넙치 비장의 NK cell과의 공배양 시간은 4시간과 18시간을 선정하여 실험을 실시하였다. YAC-1 cell을 사용하여 실험의 경우 4시간과 18시간 공배양 실험 모두 200 mg/kg 용량 구간의 실험군에서 가장 높은 세포독성을 대조군 대비 최대 3.06배 높은 세포독성을 보였다. HINAE cell을 사용한 실험의 경우 4시간 공배양한 실험에서만 유의적인 차이를 보였으며, YAC-1 cell과 마찬가지로 200 mg/kg 용량 구간의 실험군에서 가장 높은 세포 독성을 보여 대조군 대비 2.3배 높은 세포독성을 보였다. 추가적으로 넙치의 두신 조직에서 IL-12b의 발현량을 확인하였고, 세포독성 실험과 일치하는 결과를 보였고, 200 mg/kg 용량 구간의 실험군에서 가장 높은 발현량을 보여 대조군과 비교하여 6.62배 높은 수치를 보였다. 이러한 결과는 N-(4-(4-Fluorophenoxy)-3-methylphenyl) acetamide가 넙치의 NK cell 활성에 영향을 줄 수 있다는 것을 보여준다.

Keywords

Acknowledgement

이 논문은 국립수산과학원(P2023202, 넙치쿠도아충 저감화 연구)의 지원에 의해 진행되었음.

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