title: Rac1 regulates platelet shedding of CD40L in abdominal sepsis.
abstract: Matrix metalloproteinase-9 (MMP-9) regulates platelet shedding of CD40L in abdominal sepsis. However, the signaling mechanisms controlling sepsis-induced shedding of CD40L from activated platelets remain elusive. Rac1 has been reported to regulate diverse functions in platelets; we hypothesized herein that Rac1 might regulate platelet shedding of CD40L in sepsis. The specific Rac1 inhibitor NSC23766 (N6-[2-[[4-(diethylamino)-1-methylbutyl] amino]-6-methyl-4-pyrimidinyl]-2 methyl-4, 6-quinolinediamine trihydrochloride) was administered to mice undergoing cecal ligation and puncture (CLP). Levels of CD40L and MMP-9 in plasma, platelets, and neutrophils were determined by use of ELISA, western blot, and confocal microscopy. Platelet depletion abolished the CLP-induced increase in plasma levels of CD40L. Rac1 activity was significantly increased in platelets from septic animals. Administration of NSC23766 abolished the CLP-induced enhancement of soluble CD40L levels in the plasma. Moreover, Rac1 inhibition completely inhibited proteinase-activated receptor-4-induced surface mobilization and secretion of CD40L in isolated platelets. CLP significantly increased plasma levels of MMP-9 and Rac1 activity in neutrophils. Treatment with NSC23766 markedly attenuated MMP-9 levels in the plasma from septic mice. In addition, Rac1 inhibition abolished chemokine-induced secretion of MMP-9 from isolated neutrophils. Finally, platelet shedding of CD40L was significantly reduced in response to stimulation with supernatants from activated MMP-9-deficient neutrophils compared with supernatants from wild-type neutrophils, indicating a direct role of neutrophil-derived MMP-9 in regulating platelet shedding of CD40L. Our novel data suggest that sepsis-induced platelet shedding of CD40L is dependent on Rac1 signaling. Rac1 controls surface mobilization of CD40L on activated platelets and MMP-9 secretion from neutrophils. Thus, our findings indicate that targeting Rac1 signaling might be a useful way to control pathologic elevations of CD40L in the systemic circulation in abdominal sepsis.
authors.affiliation: Department of Clinical Sciences, Malmö, Section for Surgery, Lund University, Skåne University Hospital, Malmö, Sweden.
authors.forename: Rundk
authors.initials: R
authors.lastname: Hwaiz
authors.affiliation: Department of Clinical Sciences, Malmö, Section for Surgery, Lund University, Skåne University Hospital, Malmö, Sweden.
authors.forename: Milladur
authors.initials: M
authors.lastname: Rahman
authors.affiliation: Islet Pathophysiology, Lund University, Malmö, Sweden.
authors.forename: Enming
authors.initials: E
authors.lastname: Zhang
authors.affiliation: Department of Clinical Sciences, Malmö, Section for Surgery, Lund University, Skåne University Hospital, Malmö, Sweden.
authors.forename: Henrik
authors.initials: H
authors.lastname: Thorlacius
mesh_terms: D000005:Abdomen; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D023201:CD40 Ligand / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D008297:Male; D020784:Matrix Metalloproteinase 8 / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009504:Neutrophils / Q000201:enzymology; D018805:Sepsis / Q000097:blood*; D020830:rac1 GTP-Binding Protein / Q000502:physiology*
chemical_list: D023201:CD40 Ligand; D020784:Matrix Metalloproteinase 8; D020830:rac1 GTP-Binding Protein