Korean J Phys Anthropol.  2016 Mar;29(1):19-26. 10.11637/kjpa.2016.29.1.19.

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

Affiliations
  • 1Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, Korea.
  • 2College of Nursing, Hanyang University, Korea. seoyk75@hanyang.ac.kr

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.

Keyword

Mouse; Left common iliac artery; Ischemia; Reperfusion injury; Pro-inflammtory cytokine

MeSH Terms

Anesthesia, General
Animals
Chemokine CCL5
Cytokines*
Enzyme-Linked Immunosorbent Assay
Heart
Humans
Iliac Artery*
Inflammation
Interleukin-2
Interleukin-3
Interleukin-5
Interleukin-6
Ischemia*
Male
Mice*
Reactive Oxygen Species
Reperfusion Injury
Reperfusion*
Risk Factors
Chemokine CCL5
Cytokines
Interleukin-2
Interleukin-3
Interleukin-5
Interleukin-6
Reactive Oxygen Species

Figure

  • Fig. 1. The changes in the concentration of cytokine in the serum after of ischemia. Data are presented as mean (Pg/mL). Kruskal-Wallis test was performed to determine any differences between the reperfusion groups including the 0-hour reperfusion group as a control group, and the levels of all cytokines were significantly different between the reperfusion groups. Mann-Whitney test was performed for post-hoc comparisons with Bonferroni correction method, and one asterisk indicates an adjusted p-value of .005.


Reference

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