P-selectin antibody reduces hemorrhage and infarct volume resulting from MCA occlusion in the rat

doi:10.1016/S0022-510X(98)00262-7

Journal of the Neurological Sciences

Volume 161, Issue 1, 26 November 1998, Pages 16-22

https://doi.org/10.1016/S0022-510X(98)00262-7 Get rights and content

Abstract

We investigated the effect of an anti-P-selectin antibody (RMP-1) on ischemic cell damage and hemorrhage after transient middle cerebral artery occlusion (MCAo) in the rat. Animals were divided into four groups: (1) antibody (Ab) 1 group (n=14) RMP-1 (2 mg/kg) was administered to rats 1 h prior to induction of 2 h of MCA occlusion; (2) control-vehicle group Ab2 (n=12) rats were subjected to the same experimental protocol, except that an isotype-matched control antibody was administered; (3) Ab1 group (n=10) rats were subjected to 2 h of MCA occlusion and RMP-1 (2 mg/kg) was administered upon reperfusion; (4) control-vehicle group Ab2 (n=10) rats were subjected to the same experimental protocol, except that an isotype-matched control antibody was administered. Animals were sacrificed 48 h after onset of the MCAo for histological evaluation of infarction and hemorrhage, and to quantify number of neutrophils. The lesion volume was significantly smaller only in pretreated rats (RMP-1 group, 18.7±3.1%) compared to the vehicle-treated (31.6±2.6%) group (P<0.01). Total area of hemorrhage (5.94×10³±2.86×10³ μm²) in the pre MCAo RMP-1 treated group animals was significantly reduced (P<0.02) compared to the vehicle group (6.1×10⁴±3.42×10⁴ μm²), respectively. Our data demonstrate that administration of the anti-P-selectin antibody before transient focal cerebral ischemia in rat brain reduces ischemic cell damage and petechial hemorrhage.

Introduction

Cerebral reperfusion after focal cerebral ischemia brings leukocytes into the damaged tissue. These cells remove dead tissue by engulfing and lysing debris. Leukocytes also release oxygen free radicals and proteinases, which can damage normal and injured cells [3].

The selectins (E-, L-, P-selectin) are three related adhesion receptors that mediate the initial rolling and subsequent adhesion of leukocytes to the vessel wall by binding to carbohydrate ligands on apposing cells. The selectins have a calcium-dependent lectin domain and bind to carbohydrate ligands such as sialyl-Lewis^x (SLe^x). This selectin–carbohydrate interaction can be blocked in vitro by synthetic oligopeptides targeting the lectin domain of selectin [26]. Since their cloning in 1989, the selectins have been the subject of intense study because of their important role in both physiological and pathological inflammation.

The P-selectin is a glycoprotein, composed of a single polypeptide chain with molecular weight of 140 000 [12]. The P-selectin (CD62P) adhesion receptor is found in secretory granules (called Weibel-Palade bodies) of endothelial cells [20], alpha granules of resting platelets [12]and transfected neuroendocrine PC-12 cells [9]. The P-selectin translocates to the cell surface shortly after activation. In vitro stimulation of endothelial cells with thrombin or histamine causes rapid and transient expression of P-selectin on the cell surface 11, 20, where it mediates adhesion of neutrophils 8, 16. In vivo P-selectin plays a very important role in the process of neutrophil and monocyte rolling and adhesion on the vascular surface and aggregation of platelets. P-selectin is upregulated at 15 min and with a peak at 6 h after middle cerebral artery occlusion (MCAo) in rats [30]. Upregulation of P-selectin has also been detected in monkeys subjected to MCAo [22]. Upregulated P-selectin is mainly located on endothelial cells in the ischemic lesion 6, 22, 30. P-selectin may have a role in facilitating leukocyte binding to the endothelial cells thereby damaging the endothelial cell.

We have previously shown that administration of an oligosaccharide analog of SLe^x, CY 1503, reduced infarct volume as well as MPO immunoreactive cells in rats subjected to transient MCAo [31]. This study suggests that selectins play an important role in cerebral ischemic cell damage caused by leukocytes. Since SLe^x inhibits both P- and E-selectins, the effect of P- or E-selectin on ischemic cell damage could not be distinguished. In the present study we used a mouse monoclonal antibody (mAb) against rat P-selectin (RMP-1) to investigate the effect of P-selectin on parenchymal hemorrhage and infarct volume in rat brain after transient focal cerebral ischemia.

Section snippets

Materials and methods

All experimental procedures have been approved by the Care of Experimental Animals Committee of Henry Ford Hospital.

Results

Table 1 shows the physiological parameters (blood pressure, blood gases) before and after administration of antibody for all treated groups of animals. No significant differences were detected within each group in arterial blood parameters before and after injection of mAb and between groups.

The placement of the tip of the suture at the origin of the MCA was confirmed in all animals. The lesion volume of cerebral infarction was significantly reduced (P=0.0055) in the RMP-1 preocclusion group of

Discussion

In this study, we utilized a monoclonal antibody against P-selectin (RMP-1) to treat ischemic cell damage resulting from MCA occlusion. This antibody to P-selectin was administered either 1 h prior to ischemia or upon reperfusion at the 2 h time point of ischemia. Pretreatment with the anti-P-selectin antibody demonstrated a significant reduction in the volume of cerebral infarction and a reduction of incidence and total area of hemorrhage. In contrast, post treatment with the anti-P-selectin

Acknowledgements

The authors wish to thank Denice Janus for manuscript preparation. This work is supported in part by NINDS grants PO1 NS23393, RO1 NS33627 and RO1 NS34184.

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P-selectin antibody reduces hemorrhage and infarct volume resulting from MCA occlusion in the rat

Abstract

Introduction

Section snippets

Materials and methods

Results

Discussion

Acknowledgements

J Biol Chem

J Biol Chem

Thromb Res

Brain Res

Mol Brain Res

Anti-CD11b monoclonal reduces ischemic cell damage after transient focal cerebral ischemia in rat

Ann Neurol

The effect of hypothermia on the transient middle cerebral artery occlusion in the rat

J Cerebral Blood Flow Metab

Role of oxidants in ischemic brain damage

Stroke

Antibodies against adhesion molecules reduce apoptosis after transient middle cerebral artery occlusion in rat brain

J Cereb Blood Flow Metab

Postischemic administration of an anti-Mac-1 antibody reduces ischemic cell damage after transient middle cerebral artery occlusion in rats

Stroke

Exacerbation of cerebral injury in mice that express the P-selectin gene: identification of P-selectin blockade as a new target for the treatment of stroke

Circulation Res

The tree members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewisx oligosaccharide

J Cell Biol

Rapid neutrophil adhesion to activated endothelium mediated by GMP-140

Nature

The cytoplasmic domain of P-selectin contains a sorting determinant that mediates rapid degradation in lysosomes

J Cell Biol

Experimental anti-leukocyte interventions in cerebral ischemia

J Cereb Blood Flow Metab

The tree members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis^x oligosaccharide