| ObjectiveThis study was to establish the model of ischemia-reperfusion on hind limb in rabbit.We assessed the concentrations of serum IL-1, IL-6and TNF-α at different time pointsduring the occurrence and development of ischemia reperfusion on hind limb in rabbit.The aim is to study the effect of serum IL-1, IL-6and TNF-α on ischemia reperfusionof hind limb in rabbit. We hope find a better kind of approach and experimental proof toprevent and treat ischemia reperfusion injury.MethodsTwenty New Zealand white rabbits, the average weight of3kg. Male or female, wererandomly divided into2groups to compare study,sham group and ischemia-reperfusiongroup(IR).The sham group as for sham-operated. The IR group for the rabbits submittedto0h and4h before the hind limb ischemia and3,6,9,12,15h after hind limbreperfusion. Collected blood at different time points and stood for1hour,thencentrifuged at3000r/min for10min at4℃;the serum was separated and stored at-80℃for ELISA to determine the concentrations of IL-1, IL-6and TNF-α. After the rabbitswere sacrificed,3cm sections of the right hind limb muscle tissue were excised, driedon filter paper, fixed in paraformaldehyde(1:10),embedded in paraffin and cut intosections10μm in thickness. Then the sections were flattened, mounted, and heated onblank glass slides. Histological evaluations were performed by hematoxylin and eosinstaining and pathological examination.Result 1.The serum levels of IL-1, IL-6and TNF-α in IR groups were significantly higherthan those in the sham groups(P＜0.05).All the concentrations of inflammatory factorsin rabbits serum were increased with time. But there was no significant difference insham groups(P＞0.05).2.IL-1, TNF-a reached the peak at9h(P<0.05) after reperfusion, which in the shamgroups did not observe this phenomenon. And those inflammatory factors among shamgroups had no significant difference(P＞0.05).3.IL-6reached the peak at12h(P<0.05) after reperfusion,but the concentration of IL-6had no significant difference in sham groups(P＞0.05).4.Skeletal muscle morphology showed no change in the sham groups. In contrast, theskeletal muscle structure showed inflammation and muscle interstitial edema inischemia-reperfusion groups. But there was no conspicuous necrosis..Conclusion1.We successfully established the model of ischemia-reperfusion injury on rabbit hindlimb;2.Our experiment suggest that the serum inflammatory cytokines, IL-1, IL-6and TNF-αparticipate in hind limb ischemia-reperfusion injury;3Serum concentration of IL-1, IL-6, TNF-α undulate at different time points ofischemia-reperfusion, which may conduct monitoring of the occurrence anddevelopment of ischemia-reperfusion injury;4The level of serum of IL-6can reveal the severity of the tissue damage. And TNF-αcan induce the production of IL-6,the latter can stimulate chain reaction ofinflammation;5. Inflammatory cytokines, which results from ischemia-reperfusion, will damageorganized system. |
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