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International Angiology 2001 March;20(1):78-89


lingua: Inglese

Circulating neutrophil priming and systemic inflammation in limb ischaemia-reperfusion injury

Harkin D. W., Barros D’sa A. A. B., McCallion K., Hoper M., Halliday M. I., Campbell F. C.

From the Department of Surgery, The Queen’s University of Belfast * Vascular Surgical Unit, The Royal Victoria Hospital, Belfast, UK


Background. Recruitment and acti­va­tion of neu­troph­ils is a key step in the devel­op­ment of local and system­ic inju­ry in lower limb ischae­mia-reper­fu­sion. We hypo­the­sise that ­increased cir­cu­lat­ing neu­troph­il prim­ing is respon­sible for system­ic inflam­ma­tion.
Methods. Anaesthetised ven­ti­lat­ed swine (n=6 per group) under­went mid-line lap­a­rot­o­my and were ran­dom­ised to con­trol group or bilat­er­al exter­nal iliac ­artery occlu­sion for two hours fol­lowed by two and a half hours reper­fu­sion (I/R group). Using lumi­nol, res­pir­a­to­ry burst activ­ity was ­assayed with a BioOrbit Luminometer to ­detect whole blood chem­ilu­mi­nes­cence (CL) by stim­u­la­tion with phor­bol 1,2-myr­is­tate 1,3-ace­tate (PMA) in the ­absence or pres­ence of ­tumour necro­sis fac­tor (TNF) respec­tive­ly. PMN prim­ing is ­expressed as the ratio of whole blood CL in the pres­ence of TNF to that with­out. We meas­ured plas­ma inter­leu­kin(IL)-6 and ­tumour necro­sis fac­tor alpha by bio­as­say as a meas­ure of system­ic inflam­ma­tion. The alveo­lar-arte­ri­al (A-a) gra­di­ent was meas­ured using the for­mu­la [(A-a)gra­di­ent=frac­tion ­inspired O2×710-(arte­ri­al pCO2/0.8)-arte­ri­al pO2], it is a meas­ure of lung func­tion, a large gra­di­ent being indic­a­tive of ­impaired oxy­gen trans­port and hence lung inju­ry.
Results. Lower limb I/R ­caused sig­nif­i­cant­ly great­er PMN prim­ing, 0.83±0.14, com­pared to con­trol group, 0.22±0.04, (p<0.001). Plasma IL-6, a reli­able indi­ca­tor of system­ic inflam­ma­tion, was sig­nif­i­cant­ly ­increased in I/R group after two and a half hours of reper­fu­sion, 1295.0 (833.9-2073.0) pg/L, com­pared to con­trol, 382.9 (367.4-568.3) pg/L, (p<0.005). Plasma ­tumour necro­sis fac­tor alpha was sig­nif­i­cant­ly ele­vat­ed after one hour of reper­fu­sion in the I/R group, 86.8 (48.7-106.6) pg/ml, com­pared to the con­trol group, 32.7 (0.9-42.8) pg/ml, (p<0.01). (A-a) gra­di­ent was sig­nif­i­cant­ly ­increased after IRI, 407.97±53.13, com­pared to the con­trol, 183.19±45.75, (p<0.005). Mean pul­mo­nary ­artery pres­sure was sig­nif­i­cant­ly great­er after IRI, 38.80±4.87 mmHg, com­pared to con­trol, 27.86±1.92 mmHg, (p<0.005). Data rep­re­sents mean±stan­dard error mean or ­median (inter­quar­tile range), sta­tis­ti­cal com­par­i­sons using one-way Anova with Student’s “t”-test and Kruskall-Wallis Anova with the Mann-Whitney U test.
Conclusions. Priming of neu­troph­ils increas­es their cir­cu­lat­ing res­pir­a­to­ry burst activ­ity and abil­ity to ­induce tis­sue inju­ry. Systemic PMN prim­ing dur­ing hind limb ischae­mia-reper­fu­sion inju­ry is asso­ciat­ed with the system­ic inflam­ma­to­ry ­response syn­drome.

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