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2시간 허혈 후 재관류 시점에 따른 흰쥐의 혈청 및 장딴지근의 친염증성 사이토카인 및 항염증성 사이토카인의 변화

The Changes of Pro- and Anti-inflammatory Cytokines in Both Serum and Gastrocnemius Muscle of Mice after 2-hour Postischemic Reperfusion Injury

김신영
대한체질인류학회|2014.06|해부·생물인류학 (Anatomy & Biological Anthropology)|제27권 제2호|pp. 101-111|DOI 10.11637/aba.2014.27.2.101

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참고문헌

키워드

IschemiaPro-inflammatory cytokineReperfusion허혈재관류친염증성 사이토카인항염증성 사이토카인Anti-inflammatory cytokine
주제분류해부학

초록

본 연구는 허혈 후 재관류 시점에 따라 혈청과 뼈대근육에서 나타나는 친염증성 및 항염증성 사이토카인(pro- and anti-inflammatory cytokine)의 변화 양상을 비교 분석하여 뼈대근육의 허혈 후 재관류 시점에 따라 나타나는 염증성 반응의 변화와 이를 대표할 수 있는 표지인자를 확인하는 데 목적이 있다. 실험동물은 12주령 ICR계 수컷 생쥐를 사용하여 대조군과 허혈 후 재관류 시간에 따라 재관류 0, 0.5, 1, 2, 4,8, 16, 24시간군으로 분류하였고, 허혈 처치는 왼온엉덩동맥(left common iliac artery)을 2시간 동안 혈관집게로차단하였다. 재관류 시간경과에 맞추어 실험동물을 마취시킨 후 심장의 오른 심방에서 채취한 혈액의 혈청과장딴지근 조직액에서 친염증성 사이토카인 (Eotaxin, IFNγ, IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, MCP-1, MDC,MIP-1α, RANTES, TARC, TCA-3)과 항염증성 사이토카인(IL-4, IL-10)의 변화를 확인하기 위하여 ELISA 분석을 시행하였다. 연구결과 허혈 후 재관류 시점에 따른 친염증성 사이토카인의 발현은 뼈대근육 내 재관류 0시간군에서, 혈청에서는 대조군과 재관류 0, 1, 2, 4, 8, 16시간군에서 사이토카인들 사이의 발현양의 차이가 유의한 것으로 관찰되었고(p⁄.05), 항염증성 사이토카인의 발현은 혈청 내 재관류 4시간군에서 사이토카인들 사이의 발현양의 차이가 유의한 것으로 관찰되었다(p⁄.05). 또한 대조군과 비교하여 유의한 차이를 보이는 재관류 시간은 친염증성 사이토카인에서는 혈청 내 재관류 2, 4, 16시간군이었고, 항염증성 사이토카인에서는 혈청 내 재관류 4시간군이었다(p⁄.05). 본 연구를 토대로 허혈 후 사이토카인들의 변화 양상은 재관류 시점에 따라 뼈대근육보다는 혈청에서 발현양의 차이가 유의하게 나타났으며, 특히 혈청에서 재관류 4시간군에 관찰되는 IL-6와 재관류 16시간군에 관찰되는 MCP-1은 다른 사이토카인보다 유의하게 높은 농도를 보임에 따라 친염증성 사이토카인을 대표하는 표지인자가 될 수 있을 것으로 사료된다.

Abstract

This study aimed to investigate the inflammatory changes and their main indicators according to thetime-period of postischemic reperfusion injury confirmed by analyzing changes of both pro-inflammatory andanti-inflammatory cytokines in the skeletal muscle and serum. By using 12-week-old male ICR strain mice were grouped into sham control and 8 different time-periods ofreperfusion groups (0, 0.5, 1, 2, 4, 8, 16, 24 hours). Left common iliac artery of each mice in the reperfusion groupwas devascularized by a vascular clamp for 2 hours. Once anesthesia was applied to the experimental animals,blood serum was obtained from right heart atrium on the difference time-period of reperfusion (0-, 0.5-, 1-, 2-, 4-,8-hour, respectively). Then, tissue fluid was collected in calf muscles (gastrocnemius muscle) after the mice weresacrificed by cervical dislocation. By using these serum and tissue fluids, enzyme-linked immunosorbent assay(ELISA) was used to analyze both pro-inflammatory cytokines (Eotaxin, IFNγ, IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, MCP-1, MDC, MIP-1α, RANTES, TARC, TCA-3) and anti-inflammatory cytokines (IL-4, IL-10). Consequently,there were significant differences of pro-inflammatory cytokines levels in the skeletal muscle of 0-hour reperfusion group (p⁄.05) and those in the serum of 0-, 1-, 2-, 4-, 8-, 16-hour reperfusion groups (p⁄.05). In the serum of 4-hour reperfusion group, the presence of anti-iflammatory cytokines was significant from othergroups (p⁄.05). By the comparison with the control group, furthermore, pro-inflammatory cytokines in theserum of 2-, 4-, 16-hour reperfusion group and anti-inflammatory cytokines in the serum of 4-hour reperfusiongroup were considerably different (p⁄.05). To sum up, changes of cytokine levels according to the time-period of reperfusion were considerably differentin the serum rather than the tissue fluids from the skeletal muscle. In particular, IL-6 and MCP-1 in the serumshowed higher density in 4- and 16-hour reperfusion groups so that they could be considered as the mainindicator of pro-inflammatory cytokines.

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