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

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.

The Effect of Ischemic Preconditioning on the Expression of Serum Cytokines after 2 Hours Ischemia and Timely Reperfusion in the Hindlimb of Mice / 체질인류학회지

The large volume of reactive oxygen species are generated during reperfusion after transient or post-procedural ischemia, which leads to cell injury. This ischemia-reperfusion (IR) injury may cause local and even systemic injuries. Thus, the need to reduce the IR injury has been highlighted and in this regard studies have demonstrated the ischemic preconditioning (IP) in which short ischemia and reperfusion are repeated before ischemia. Such IP is known to protect the tissues from IR injury by reducing inflammation response during ischemia. Thus, this study was based on IP known to protect the tissue with developing the mechanism of resistance to ischemia and reperfusion injury in cellular tissue. As the substance that plays an important role in the inflammatory response during IR injury is cytokines, this study was intended to review and discuss the methodologies of IP as well as to analyze the correlation of its effects on the expression of cytokines. Left common iliac artery in male mice of which weight was from 40 g to 45 g, was treated for ischemia. The animal groups consisted of ischemia (IC) group receiving 2-hour ischemic treatment alone; IP group receiving short 5-minute ischemia and reperfusion treatments repeated three times; and, ischemic preconditioning-ischemia (IP-IC) group receiving IP treatment followed by 2-hour ischemic treatment. Following these treatments in each group, reperfusion for intergroup comparisons was carried out at 30 minutes, and 1, 2 and 4 hours. The results of this study were as follows: First, the expression level of pro-inflammatory cytokines, IL-1β was the highest in IC group receiving 2-hour ischemic treatment alone (p<.001). Second, the expression level of anti-inflammatory cytokine, IL-4 was the highest in the IP group (p<.001). Third, the expression level of anti-inflammatory cytokine, IL-10 was the highest in the IP group (p<.001). In conclusion, even though the results had the degree of difference, the expression level of pro-inflammation cytokine, IL-1β in IC group was significantly lower than that in IP group, and the expression levels of anti-inflammatory cytokines, IL-4 and IL-10 in IP were significantly higher than those in IC group and IP-IC group.