Korean J Phys Anthropol.  2014 Jun;27(2):101-111. 10.11637/kjpa.2014.27.2.101.

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

Affiliations
  • 1Department of Anatomy and Cell Biology, Hanyang University, Korea. 72shin02@hanmail.net

Abstract

This study aimed to investigate the inflammatory changes and their main indicators according to the time-period of postischemic reperfusion injury confirmed by analyzing changes of both pro-inflammatory and anti-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 of reperfusion groups (0, 0.5, 1, 2, 4, 8, 16, 24 hours). Left common iliac artery of each mice in the reperfusion group was 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 were sacrificed by cervical dislocation. By using these serum and tissue fluids, enzyme-linked immunosorbent assay (ELISA) was used to analyze both pro-inflammatory cytokines (Eotaxin, IFNgamma, IL-1alpha, IL-1beta, IL-2, IL-3, IL-5, IL-6, MCP-1, MDC, MIP-1alpha, 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 other groups (p<.05). By the comparison with the control group, furthermore, pro-inflammatory cytokines in the serum of 2-, 4-, 16-hour reperfusion group and anti-inflammatory cytokines in the serum of 4-hour reperfusion group were considerably different (p<.05). To sum up, changes of cytokine levels according to the time-period of reperfusion were considerably different in the serum rather than the tissue fluids from the skeletal muscle. In particular, IL-6 and MCP-1 in the serum showed higher density in 4- and 16-hour reperfusion groups so that they could be considered as the main indicator of pro-inflammatory cytokines.

Keyword

Ischemia; Reperfusion; Pro-inflammatory cytokine; Anti-inflammatory cytokine

MeSH Terms

Anesthesia
Animals
Chemokine CCL3
Chemokine CCL5
Cytokines*
Dislocations
Enzyme-Linked Immunosorbent Assay
Heart Atria
Humans
Iliac Artery
Interleukin-2
Interleukin-3
Interleukin-5
Interleukin-6
Ischemia
Male
Mice*
Muscle, Skeletal*
Muscles
Reperfusion
Reperfusion Injury*
Serum
Chemokine CCL3
Chemokine CCL5
Cytokines
Interleukin-2
Interleukin-3
Interleukin-5
Interleukin-6
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