Upregulated Expression of Toll-like Receptor 4 in Peripheral Blood of Ischaemic Stroke Patients Correlates with Cyclooxygenase 2 Expression

AbstractObjectivesAn inflammatory process following stroke in human brains and systemic inflammatory responses after stroke in humans have been reported by numerous investigators. The aim of the study was to investigate if genes involved in the cyclooxygenase 2 (COX-2) pathway are upregulated at peripheral level in patients after transient ischaemic attack (TIA) and stroke.Design of StudyBlood samples were obtained from two groups of patients undergoing carotid endarterectomy. The first group included 25 patients who presented TIA or ischaemic stroke. The second group included 35 patients who had an asymptomatic internal carotid artery stenosis. Total RNA was isolated and the expression of Toll-like Receptor 4 (TLR4), COX-2, membrane-associated Prostaglandin E synthase (mPGES-1), Prostaglandin E2 receptors (EP3 and EP4) was analysed by real time RT-PCR.ResultsExpression of COX-2 and TLR4 were significantly increased in symptomatic patients (p<0.001). Correlation analysis showed that TLR4 expression significantly correlated with COX-2 expression (R=0.65; p<0.01) in ischaemic stroke patients. This correlation was not observed in TIA and asymptomatic patients.ConclusionsOur results suggest that the peripheral mechanism of inflammatory injury after stroke may be mediated by TLR4 through a COX-2-dependent pathway

Cyclooxygenase 2, toll-like receptor 4 and interleukin 1beta mRNA expression in atherosclerotic plaques of type 2 diabetic patients.

Objectives and design: Inflammation has a prominent role in the development of atherosclerosis. Type 2 diabetes could contribute to atherosclerosis development by promoting inflammation. This status might accelerate changes in intrinsic vascular wall cells and favor plaque formation. Cyclooxygenase 2 (COX-2) is highly expressed in atherosclerotic plaques. COX-2 gene expression is promoted through activation of toll-like receptor 4 (TLR4) and pro-inflammatory cytokine interleukin 1\u3b2 (IL1-\u3b2). Aim of this study is to investigate whether expression profiles of pro-inflammatory genes such as COX-2, TLR4 and IL1-\u3b2 in atherosclerotic plaques are altered in type 2 diabetes (T2D).Methods: Total RNA was isolated from plaques of atherosclerotic patients and expression of COX-2, TLR4, IL1-\u3b2 analyzed using real-time PCR. Histological analysis was performed on sections of the plaque to establish the degree of instability.Results: Statistically significant differences in mRNA expression of COX-2 and IL1-\u3b2 were found in plaques of T2D compared with non-T2D patients. A multi-variable linear regression model suggests that COX-2 mRNA expression is affected by T2D pathology and IL1-\u3b2 mRNA expression in atherosclerotic plaques.Conclusions: Our results support the hypothesis that T2D pathology contributes in vivo to increase the inflammatory process associated with the atherosclerotic plaque formation, as shown by an increment of COX-2 and IL1-\u3b2 mRNA expression

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