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왼온엉덩동맥 허혈 후 재관류 시간경과에 따른 생쥐의 혈청 내친염증성 cytokine의 변화

The Changes of Pro-inflammatory Cytokines in Serum according to the Reperfusion Time after Ischemia of Left Common Iliac Artery in Mice

김성재|서윤경|박경신
대한체질인류학회|2016.03|해부·생물인류학 (Anatomy & Biological Anthropology)|제29권 제1호|pp. 19-26|DOI 10.11637/aba.2016.29.1.19

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참고문헌

키워드

IschemiaMouse생쥐허혈왼온엉덩동맥재관류 손상친염증성 cytokineLeft common iliac arteryReperfusion injuryPro-inflammtory cytokine
주제분류해부학

초록

허혈 재관류 손상은 허혈이 이루어진 후 재관류되면서 일어난다. 허혈 재관류 손상의 주요 원인은 활성산소기와 활성화된 면역세포가 분비하는 cytokine이다. Cytokine은 감염이나 면역반응, 염증 등의 반응을 조절하는 세포 사이의 신호전달물질로, 염증반응을 증가시키는 것을 친염증성 cytokine이라 한다. 본 실험에서 저자들은 허혈후 재관류 경과에 따른 친염증성 cytokine인 IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, eotaxin, MCP-1, MDC, MIP-1α, RANTES, TARC, IFNγ의 변화를 알아보았다. 12주령의 수컷 생쥐를 이용하여 정상대조군과 2시간 허혈군, 4시간 허혈군, 6시간 허혈군으로 구분하였고, 총 96마리의 생쥐를 사용하였다. 허혈처치는 왼온엉덩동맥을 혈관집게를 이용하여 차단하였고, 각 허혈군은 재관류 경과에따라 0시간, 2시간, 4시간, 8시간, 16시간으로 재분류하였다. 재관류 시간에 따라 생쥐를 마취시킨 후 심장에서 혈액을 채취하였다. 혈청을 분리한 후 ELISA 분석방법을 이용하여 IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, eotaxin, MCP-1, MDC, MIP-1α, RANTES, TARC, IFNγ의 농도를 측정하여 다음과 같은 결과를 얻었다. 13개의 cytokine 농도는 허혈시간에 따라 그리고 재관류 시간에 따라 유의한 차이가 있었다. 허혈 2시간군에서IL-1α와 IL-3는 정상대조군보다 유의성 있게 증가되었고, 재관류 후 MIP-1α를 제외한 모든 cytokine들은 증가하였으며, MCP-1과 TARC은 재관류 16시간에 가장 높은 농도로 관찰되었다. 허혈 4시간군에서 TARC만이 정상대조군보다 유의성 있게 증가되었으며, 재관류 후 모든 cytokine들은 재관류 4시간 이후에 감소하는 경향으로 관찰되었다. 허혈 6시간군에서 IL-2, IL-3, MCP-1, TARC은 정상대조군보다 유의성 있게 증가되었고, 재관류 16시간에 IL-3와MCP-1이 유의성 있게 증가되었다. 이상의 결과를 종합하면, 허혈은 친염증성 cytokine을 정상대조군보다 증가시키는 것을 알 수 있었고, 특히 2시간과 6시간 허혈군에서 관찰된 IL-1α, IL-3, MCP-1, TARC은 재관류 후기까지 비교적 높은 농도로 유지되는 것을 알수 있었다.

Abstract

Ischemia-reperfusion injury arises from the restoration of blood supply after ischemia. Both reactive oxygen species and various cytokines produced by activated immune cells are the primary causal risk factors for ischemic injury. Cytokines are intercellular signaling substances for regulating any infection, immune reactions and inflammation, and pro-inflammatory cytokines adversely affect any diseases through an increase in inflammatory reaction. This study was conducted to investigate whether the periods of reperfusion after ischemia result in any changes of pro-inflammatory cytokines in the serum, including IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, Eotaxin, MCP-1, MDC, MIP-1α, RANTES, TARC, IFNδ. A total of 96 male mice aged at 12 weeks was used in this study, and the groups of ischemia were divided into the following three different groups: 2-hour, 4-hour, and 6-hour ischemia groups. For the object of ischemic injury, the left common iliac artery was clamped by vascular clamp, each ischemia group was subdivided into 5 different groups according to the periods of reperfusion: 0-, 2-, 4-, 8-, and 16-hour reperfusion time. Blood samples after general anesthesia were collected from the mice hearts, and the serum was separated from them. The concentration of pro-inflammatory cytokines (IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, Eotaxin, MCP-1, MDC, MIP-1α, RANTES, TARC, IFNδ) in the serum was measured by ELISA, and the following results were acquired. The concentrations of the 13 pro-inflammatory cytokines were significantly different in accordance with the periods of ischemia and the reperfusion time. In 2-hour ischemia group, IL-1α and IL-3 were increaed compared to normal control group, and 12 cytokines were increased followed by reperfusion except for MIP-1α. MCP- 1 and TARC were expressed as the highest concentration in the 16-hour reperfusion time. In 4-hour ischemia group, TARC was significant differences with normal control group, and the concentration of 13 cytokines were decreased after 4-hour reperfusion time. In 6-hour ischemia group, IL-2, IL-3, MCP-1 and TARC were increased, compared to normal control group, and IL-3 and MCP-1 were increased in 16-hour reperfusion time. To sum up, ischemia increased the pro-inflammatory cytokines compared to normal control group and in the 2-hour and 6-hour ischemia groups, IL-1α, IL-3, MCP-1 and TARC were increased until the late reperfusion time.

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