Heparin-Protamine does not Aggravate Local LPS-Provoked Leukocytic Inflammation in Vivo

Abstract

Objective: Secondary complications involving inflammation limit postoperative results in cardiac surgery. Because heparin-protamine can elicit inflammatory reactions, this study evaluates in vivo whether treatment with heparin-protamine aggravates local endotoxin-induced injury. Methods: Mice received intravenous injections of either heparin-protamine, protamine alone or PBS for controls, before local air pouch challenge with LPS. Leukocytes recruited within the air pouches were collected and analyzed by flow cytometry. Results: LPS provoked a local leukocytic infiltration in a dose- and time-dependent manner with significantly elevated numbers of 1.75 ± 0.29 × 10⁶ cells after four hours compared to non-LPS-stimulated controls (0.55 ± 0.08 × 10⁶ cells). Recruited cells comprised of 74 ± 4 % PMNLs and 26 ± 4 % MNLs. The largest fraction of MNLs was positive for the T cell-specific marker CD90.2 (59 ± 6 %). B cells were only rarely observed (4 ± 1 %). In non-LPS-challenged air pouches, heparin-protamine provoked a leukocytic infiltration, which was comparable to that observed after LPS (1.51 ± 0.22 × 10⁶ cells). However, neither heparin-protamine nor protamine alone aggravated the LPS-mediated leukocyte recruitment (2.25 ± 0.25 × 10⁶ and 1.77 ± 0.23 × 10⁶ cells). Neither treatment influenced the distribution of leukocyte subpopulations compared to PBS-treated controls. Furthermore, surface expression of CD11a and CD11b on blood leukocytes did not differ between the groups, indicating that protamine does not increase the activation of circulating leukocytes during LPS-induced local inflammation. Conclusions: Our data indicate that heparin-protamine, although pro-inflammatory in nature, does not aggravate local inflammation provoked by LPS. Thus, enhanced inflammation during the perioperative course of cardiac surgery patients seems not to be attributable to the intraoperative use of heparin-protamine.

Key words

Cardiovascular surgery - myocardial protection - coronary bypass surgery

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