Immunohistochemical study for the expression of leukocyte adhesion molecules, and FGF23 and ACE2 in P. gingivalis LPS-induced diabetic nephropathy

Objective The present study aims to examine the expression of leukocyte adhesion molecules and renal metabolic factors in diabetic mouse kidneys with periodontal pathogen Pg-LPS-induced nephropathy. Background We recently reported that the glomerular endothelium expresses toll-like receptor (TLR)2 and TLR4 in diabetic environments and TLR2/4 ligand Porphyromonas (P.) gingivalis lipopolysaccharides (Pg-LPS) induce nephropathy in diabetic mice. It is thought that Pg-LPS promotes the chronic inflammation with the overexpression of leukocyte adhesion molecules and renal-specific metabolic enzymes by the recognition of Pg-LPS via TLR in the diabetic kidneys. There have been no reports of the effects of periodontopathic bacteria on the expression of leukocyte adhesion molecules and the accumulation of physiologically active substances in the kidney. Methods The immunohistochemical investigation was performed on diabetic mouse kidney with Pg-LPS-induced nephropathy with glomerulosclerosis in glomeruli. Results There were no vessels which expressed vascular cell adhesion molecule-1 (VCAM-1), E-selectin, or fibroblast growth factor (FGF) 23 in streptozotocin (STZ)-induced diabetic ICR mice (STZ-ICR), or in healthy ICR mice administered Pg-LPS (LPS-ICR). However, in diabetic ICR mouse kidneys with Pg-LPS-induced nephropathy (LPS-STZ) the expression of VCAM-1 and the accumulation of FGF23 were observed in renal tubules and glomeruli, and the expression of E-selectin was observed in renal parenchyma and glomeruli. The angiotensin-converting enzyme 2 (ACE2) was detected in the proximal tubules but not in other regions of ICR, STZ-ICR, or LPS-ICR. In LPS-STZ ACE2 was detected both in renal tubules as well as in glomeruli. The Mac-1 and podoplanin-positive cells increased in the renal parenchyma with diabetic condition and there was the distribution of a large number of Mac-1-positive cells in LPS-STZ. Conclusions The Pg-LPS may induce diabetic renal inflammation such as glomerulosclerosis and tubulitis with infiltration of Mac-1/podoplanin positive macrophages via glomerular overexpression of VCAM-1 and E-selectin, resulting in accumulation of both ACE2 and FGF23 which were unmetabolized with the inflammation-induced kidney damage under the diabetic condition. Periodontitis may be a critical factor in the progress of nephropathy in diabetic patients.

Induction of leukocyte adhesion molecules and renal physiologically active molecules in Porphyromonas gingivalis lipopolysaccharide-induced diabetic nephropathy

Background We recently reported that the glomerular endothelium expresses toll-like receptor (TLR)2 and TLR4 in diabetic environments and established that the TLR2 ligand Porphyromonas (P.) gingivalis lipopolysaccharides (LPS) induces nephropathy in diabetic mice. It is thought that P. gingivalis LPS promotes the chronic inflammation with the overexpression of leukocyte adhesion molecules and renal-specific metabolic enzymes by the recognition of P. gingivalis LPS via TLR in the diabetic kidneys. The present study aims to examine the expression of leukocyte adhesion molecules and renal metabolic factors in mouse kidneys with periodontal pathogen P. gingivalis LPS-induced diabetic nephropathy that was recently established. Methods The immunohistochemical investigation was performed on mouse kidney with P. gingivalis LPS-induced diabetic nephropathy model with glomerulosclerosis in glomeruli. Results There were no vessels which expressed vascular cell adhesion molecule-1 (VCAM-1), E-selectin, or fibroblast growth factor (FGF) 23 in diabetic mice, or in healthy mice administered P. gingivalis LPS. However, in diabetic mouse kidneys with P. gingivalis LPS-induced nephropathy the expression of VCAM-1 and the accumulation of FGF23 were established in renal tubules and glomeruli, and the expression of E-selectin was established in renal parenchyma and glomeruli. The angiotensin-converting enzyme 2 (ACE2) was detected in the proximal tubules but not in other regions including not in distal tubules of diabetic mice without LPS, and not in healthy mice administered P. gingivalis LPS. In diabetic mouse kidneys with P. gingivalis LPS-induced nephropathy ACE2 was detected both in renal tubules as well as in glomeruli. The macrophage-1 (Mac-1) and podoplanin-positive cells increased in the renal parenchyma with diabetic condition and there was accumulation in P. gingivalis LPS-induced diabetic nephropathy. As the expression of VCAM-1 and E-selectin is upregulated in glomeruli, tubules, and intertubular capillaries, it is thought that the inflammatory infiltration of the monocyte-macrophage lineage promoted in kidneys with P. gingivalis LPS-induced the diabetic nephropathy. Conclusions P. gingivalis LPS may progressively accelerate the development of the renal inflammatory environment in LPS-accumulated glomeruli with the macrophage infiltration via the renal expression of VCAM-1 and E-selectin, and with ACE2 overexpression and FGF23 accumulation. Periodontitis may be a critical factor in the progress of nephropathy in diabetic patients.

Potential role of Lu/BCAM in HIV-related atherosclerosis

Atheromatous lesions are formed by macrophages and low-density lipoprotein cholesterol invading the vascular intima. Here we show that increasing cholesterol levels are associated with peripheral monocyte depletion and this imbalance is aggravated by carriage of Lu/BCAM leukocyte adhesion molecules. This is true only in HIV infection and probably explains the risk of atherosclerosis observed in HIV-positive patients.

Direct peritoneal resuscitation reduces inflammation in the kidney after acute brain death

Brain death is associated with significant inflammation within the kidneys, which may contribute to reduced graft survival. Direct peritoneal resuscitation (DPR) has been shown to reduce systemic inflammation after brain death. To determine its effects, brain dead rats were resuscitated with normal saline (targeted intravenous fluid) to maintain a mean arterial pressure of 80 mmHg; DPR animals also received 30 cc of intraperitoneal peritoneal dialysis solution. Rats were euthanized at 0, 2, 4, and 6 h after brain death. Pro-inflammatory cytokines were measured using ELISA. Levels of IL-1β, TNF-α, and IL-6 in the kidney were significantly increased as early as 2 h after brain death and significantly decreased with DPR. Levels of leukocyte adhesion molecules ICAM and VCAM increased after brain death and were decreased with DPR (ICAM 2.33 ± 0.14 vs. 0.42 ± 0.04, P = 0.002; VCAM 82.6 ± 5.8 vs. 37.3 ± 1.9, P = 0.002 at 4 h) as were E-selectin and P-selectin (E-selectin 25,605 vs. 16,144, P = 0.005; P-selectin 82.5 ± 3.3 vs. 71.0 ± 2.3, P = 0.009 at 4 h). Use of DPR reduces inflammation and adhesion molecule expression in the kidneys, and is associated with reduced macrophages and neutrophils on immunohistochemistry. Using DPR in brain dead donors has the potential to reduce the immunologic activity of transplanted kidneys and could improve graft survival.

Endothelial Dysfunction and Inflammation: Immunity in Rheumatoid Arthritis

Inflammation, as a feature of rheumatoid arthritis (RA), leads to the activation of endothelial cells (ECs). Activated ECs induce atherosclerosis through an increased expression of leukocyte adhesion molecules. Endothelial dysfunction (ED) is recognized as a failure of endothelial repair mechanisms. It is also an early preclinical marker of atherosclerosis and is commonly found in RA patients. RA is now established as an independent cardiovascular risk factor, while mechanistic determinants of ED in RA are still poorly understood. An expanding body of study has shown that EC at a site of RA is both active participant and regulator of inflammatory process. Over the last decade, a role for endothelial dysfunction in RA associated with cardiovascular disease (CVD) has been hypothesized. At the same time, several maintenance drugs targeting this phenomenon have been tested, which has promising results. Assessment of endothelial function may be a useful tool to identify and monitor RA patients.

Abstract 12935: Therapeutic Silencing of Leukocyte Adhesion Molecules in Atherosclerosis and Acute Myocardial Infarction

INTRODUCTION/HYPOTHESIS: Leukocytes are a major component of atherosclerotic plaques, drive plaque progression and are continuously recruited from the blood. The leukocyte adhesion molecules VCAM-1, ICAM-1/2 and E-/P-Selectin are expressed on plaque endothelial cells and are the key players in this recruitment process. We here present a novel therapeutic approach that targets leukocyte recruitment to plaques. METHODS: We tested a nanoparticle system that effectively delivered short interfering RNAs targeting VCAM-1, ICAM-1/2 and E-/P-Selectin simultaneously (siCAM5) to endothelial cells of atherosclerosis prone ApoE-/- mice and C57BL/6J mice after myocardial infarction (permanent coronary ligation) and assessed leukocyte recruitment/content and plaque phenotype by flow cytometry, histology, qPCR and fluorescence molecular tomography/computed tomography imaging. RESULTS: Efficient degradation of VCAM-1, ICAM-1/2 and E-/P-Selectin on endothelial cells attenuated neutrophil and monocyte numbers in infarcts and atherosclerotic plaques (neutrophil numbers, siControl 10,974±1,250 vs. siCAM5 6,963±1,077, p<0.05; Ly6Chigh monocyte numbers, siControl 6,044±1,212 vs. siCAM5 3,770±293, p<0.05), reduced levels of pro-inflammatory cytokines in plaques, decreased protease activity in plaques (protease activity in pmol, siControl 49.4±2.8 vs. siCAM5 35.9±3.0, p<0.01), and resulted in a less inflammatory plaque phenotype (necrotic core mm2/aortic root, siControl 0.09±0.01 vs. siCAM5 0.05±0.01, p<0.05; fibrous cap μm/aortic root, siControl 36.7±9.5 vs. siCAM5 63.1±5.4, p<0.05) and lower numbers of plaque macrophages (siControl 30,313±4,414 vs. siCAM5 17,198±2,070, p<0.01). CONCLUSIONS: Taken together, the described approach can successfully suppress leukocyte recruitment to atherosclerotic plaques and infarcted hearts. It may allow aggressive medical interventions in patients with inflammatory atherosclerosis.

Rosiglitazone Influences the Expression of Leukocyte Adhesion Molecules and CD14 Receptor in Type 2 Diabetes Mellitus Patients

Diabetes mellitus is associated with increased inflammatory response, which may contribute to atherosclerosis progression. Experimental results demonstrated anti-inflammatory activity of glitazones; their effect on leukocyte adhesion molecules has not been studied to date. We therefore studied the effect of rosiglitazone treatment on leukocyte surface expression of adhesion molecules in patients with type 2 diabetes mellitus and compared our results with findings in healthy subjects. 33 subjects with type 2 diabetes and 32 healthy controls were included; patients were examined at baseline and after 5 months of rosiglitazone treatment (4 mg/d). Leukocyte expression of adhesion molecules LFA-1, CD18 and ICAM-1 was quantified using flow cytometry; in addition, CD14 (lipopolysaccharide receptor) expression was analyzed as a marker of nonspecific immunity. The expression of examined molecules at baseline was higher in patients compared to controls. Despite only mild decrease in blood glucose, rosiglitazone treatment induced substantial decrease of CD18 and CD14 expression and borderline decrease of LFA-1 and ICAM-1 expression (on monocytes only). We thus observed improvement in the expression of leukocyte inflammatory markers after rosiglitazone treatment. This effect is supposed to be mediated by direct effect of rosiglitazone on PPAR-γ receptors on leukocytes.