Bong Seon Kim

Profile of Fos-like immunoreactivity induction by light stimuli in the intergeniculate leaflet is different from that of the suprachiasmatic nucleus.

Light stimuli induce Fos-like immunoreactivity (FLI) in suprachiasmatic nucleus (SCN) and intergeniculate leaflet (IGL). Short pulses of light stimuli that synchronize the circadian rhythms induce FLI in SCN. The characteristics of light induction of FLI in the IGL were studied using immunohistochemistry. In the IGL, at least 2 h of sustained light stimuli were necessary to show an increase of FLI. This FLI persisted while the light was turned on. FLI induction in the IGL by light stimuli was not circadian time specific response. These findings imply that the functional significance of Fos activation on circadian rhythms and mechanism of FLI induction in IGL would be different from that in SCN.

Chloroform extract of aged black garlic attenuates TNF-α-induced ROS generation, VCAM-1 expression, NF-κB activation and adhesiveness for monocytes in human umbilical vein endothelial cells

Aged black garlic is a type of fermented garlic (Allium sativum) which has been used in Oriental countries for a long time because of various biological properties of garlic derivatives. The current study explored the potential of the chloroform extract of aged black garlic (CEABG) in attenuating the activities of adhesion molecules in tumor necrosis factor‐α (TNF‐α)‐stimulated human umbilical vein endothelial cells (HUVECs). The study was performed on HUVECs that were pretreated with 30 μg/mL of CEABG before TNF‐α treatment. Treatment of HUVECs with CEABG significantly inhibited TNF‐α‐induced reactive oxygen species (ROS) formation. HUVECs treated with CEABG showed markedly suppressed TNF‐α‐induced mRNA expression of VCAM‐1, but little alteration in ICAM‐1 and E‐selectin mRNA expression. CEABG treatment also significantly decreased the TNF‐α‐induced cell surface and total protein expression of VCAM‐1 without affecting ICAM‐1 and E‐selectin expression. In addition, treatment of HUVECs with CEABG markedly reduced THP‐1 monocyte adhesion to TNF‐α‐stimulated HUVECs. Furthermore, CEABG significantly inhibited NF‐κB transcription factor activation in TNF‐α‐stimulated HUVECs. The data provide new evidence of the antiinflammatory properties of CEABG that may have a potential therapeutic use for the prevention and treatment of vascular diseases such as atherosclerosis through mechanisms involving the inhibition of VCAM‐1 expression and NF‐κB activation in vascular endothelial cells. Copyright

MECHANISM OF REDUCED GFR IN RABBITS WITH ISCHEMIC ACUTE RENAL FAILURE

A reduction in glomerular filtration rate (GFR) is a primary characteristic of ischemic acute renal failure. The present study was undertaken to examine the roles of angiotensin II, tubuloglomerular-feedback (TGF) mechanism, and tubular obstruction for the GFR reduction in the postischemic kidney. Renal ischemia was induced by occlusion of the bilateral renal arteries for 60 min, and renal function was examined at 2 and 24 h after the onset of reflow. After the end of 2-h reflow, the GFR was not significantly changed, but the urine flow increased significantly. On the other hand, at the end of 24-h reflow, the GFR and urine flow decreased markedly along with increased filtration fraction. The renal blood flow significantly decreased at 24 h, but not 2 h, after reflow, which was accompanied by increased total renal vascular resistance. Furosemide infusion (1 mg/min/kg) after 24 h of reflow prevented the reduction in GFR and filtration fraction without no changes in renal blood flow and total renal vascular resistance. Pretreatment of enalapril and losartan did not prevent the reduction in GFR, indicating that angiotensin II was not involved. In morphological examinations, tubular obstruction was seen in the proximal and distal tubules of kidneys both at 2 and 24 h after the onset of reflow. In two rabbits subjected to 48 h of reflow, the tubular obstruction was not observed, despite GFR remained depressed. These results suggest that the late reduction in GFR in postischemic kidneys is not mediated by angiotensin II, but is mediated, at least in part, by the TGF mechanism. The tubular obstruction may be not prerequisite for the GFR reduction in rabbits.

Developmental expression of ‘RZRβ, a putative nuclear-melatonin receptor’ mRNA in the suprachiasmatic nucleus of the rat

RZR beta is a member of the retinoid Z receptor (RZR) family of orphan receptors, and its expression is brain-specific. Recently, it was reported that the distribution of RZR beta mRNA is partially coincident with melatonin binding sites and that RZR beta is a putative nuclear receptor of melatonin. Using in situ hybridization, we investigated the developmental expression of RZR beta mRNA in the suprachiasmatic nucleus (SCN) of the rat, which is considered as circadian pacemaker. The RZR beta mRNA was first found in the dorsomedial portion of the SCN at embryonic day 20, and RZR beta mRNA expression in the dorsomedial SCN continued until postnatal day 60. This result suggests that RZR beta plays specific roles as a transcription factor in the dorsomedial SCN but not in the ventrolateral SCN during development and throughout postnatal life.

EFFECT OF PENTOXIFYLLINE ON ISCHEMIC ACUTE RENAL FAILURE IN RABBITS

Previous studies have demonstrated that levels of tumor necrosis factor-α (TNF-α) or its mRNA expression are increased in acute renal failure of various types including ischemia/reperfusion injury. This study was undertaken to determine whether pentoxifylline (PTX), an inhibitor of TNF-α production, provides a protective effect against ischemic acute renal failure in rabbits. Renal ischemia was induced by clamping bilateral renal arteries for 60 min. Animals were pretreated with PTX (30 mg/kg, i.v.) 10 min before release of clamp. At 24 h of reperfusion of blood after ischemia, changes in renal function, renal blood flow, and the expression of TNF-α mRNA were evaluated. Ischemia/reperfusion caused a marked reduction in GFR, which was accompanied by an increase of serum creatinine levels. Such changes were significantly attenuated by PTX pretreatment. PTX ameliorated the impairment of renal tubular function, but it had no effect on the reduction of renal blood flow induced by ischemia/reperfusion. The protective effect of PTX on functional changes was supported by morphological studies. The impairment of glucose and phosphate reabsorption in postischemic kidneys was associated with a depression in the expression of Na+-glucose and Na+-Pi transporters. The expression of TNF-α mRNA was increased after reperfusion, which was inhibited by PTX pretreatment. The PTX pretreatment in vitro prevented the release of lactate dehydrogenase induced by an oxidant t-butylhydroperoxide in rabbit renal cortical slices, but it did not produce any effect on the oxidant-induced lipid peroxidation, suggesting that PTX protection is not resulted from its antioxidant action. These results suggest that PTX may exert a protective effect against ischemic acute renal failure by inhibiting the production of TNF-α in rabbits.

Calcitonin gene-related peptide-like immunoreactive (CGRPI) elements in the circadian system of the mouse: an immunohistochemistry combined with retrograde transport study.

This study was based on immunohistochemical detection of calcitonin gene-related peptide-like immunoreactive (CGRPI) neurons and fibers in the suprachiasmatic nucleus (SCN) and intergeniculate leaflet (IGL) of the mouse. CGRPI neurons and fibers were found within the ventrolateral part of the SCN, in the whole extent of IGL and sparsely distributed in ventral lateral geniculate body. The presence of CGRPI structures in the SCN and IGL of the mouse further supports the hypothesis of differences in the content of neuroactive substances in the circadian clock between mammalian species. Fluorogold retrograde transport combined with CGRP immunofluorescence demonstrates that CGRPI neurons in the IGL constitute a part of IGL reciprocal connections.

Immunohistochemical Characterization of Macrophage and Dendritic Cell Subpopulations of the Spleen, Thymus, Tongue and Heart in Cyclophosphamide‐induced Immunosuppressed Rat

This study was undertaken to investigate the immunohistochemical characterization of different subpopulations of macrophages and dendritic cells (DCs) of the spleen, thymus, tongue and heart in cyclophosphamide (CY)‐induced immunosuppressed rat. After CY treatment, remarkably, ED1+, ED2+ and ED3+ macrophage subpopulations, in general, exhibited signs of cellular activation such as an increase in number and size of cell, and an upregulation of the ED1, ED2 and ED3 reactive surface molecule expression in all the organs studied, except for some macrophage subpopulations including ED1+ macrophages in the non‐lymphoid tissues. Subpopulations of DCs showed a differential sensitivity to CY. Lymphoid DCs were more sensitive to CY than non‐lymphoid interstitial DCs. CY induced a conspicuous upregulation of intercellular adhesion molecule‐1 (ICAM‐1) expression in the vascular endothelial cells, splenic marginal zone and thymic cortex. In this study, we demonstrated the in vivo effects of CY treatment on subpopulations of macrophages and DCs as well as on ICAM‐1 expression in the rat spleen, thymus, tongue and heart. Moreover, our results shed more light on the activation effects of CY on certain subpopulations of macrophages, on the differential sensitivity of DCs to CY between the immature and mature ones, on the functional role of different subpopulations of macrophages, and on the significance of upregulated ICAM‐1 expression in the splenic marginal zone and thymic cortex after CY treatment.

RANKL stimulates proliferation, adhesion and IL-7 expression of thymic epithelial cells

In many clinical situations which cause thymic involution and thereby result in immune deficiency, T cells are the most often affected, leading to a prolonged deficiency of T cells. Since only the thymic-dependent T cell production pathway secures stable regeneration of fully mature T cells, seeking strategies to enhance thymic regeneration should be a key step in developing therapeutic methods for the treatment of these significant clinical problems. This study clearly shows that receptor activator of NF-κB ligand (RANKL) stimulates mouse thymic epithelial cell activities including cell proliferation, thymocyte adhesion to thymic epithelial cells, and the expression of cell death regulatory genes favoring cell survival, cell adhesion molecules such as ICAM-1 and VCAM-1, and thymopoietic factors including IL-7. Importantly, RANKL exhibited a significant capability to facilitate thymic regeneration in mice. In addition, this study demonstrates that RANKL acts directly on the thymus to activate thymus regeneration regardless of its potential influences on thymic regeneration through an indirect or systemic effect. In light of this, the present study provides a greater insight into the development of novel therapeutic strategies for effective thymus repopulation using RANKL in the design of therapies for many clinical conditions in which immune reconstitution is required.

Wnt7b is Upregulated in Macrophages during Thymic Regeneration and Negatively Regulated by RANKL

Thymus can regenerate to its normal mass within 14 days after acute involution induced by cyclophosphamide (CY) in adult rat. Despite the established role of Wnt pathways in the process of thymus development, they have not yet been associated with the regeneration of adult thymus. The purpose of this study was to investigate whether Wnt7b, which is expressed in developing thymic epithelial cells rather than in thymocytes, is modulated during thymic regeneration in adult rat. Here, we show that Wnt7b expression was up-regulated in the regenerating thymus. Cells immunolabeled for the Wnt7b were identified as macrophages. Furthermore, Wnt7b gene expression was decreased by the treatment of receptor activator of NF-kappaB ligand (RANKL). Taken together, our results demonstrate that Wnt7b gene expression was increased in macrophages during thymic regeneration and negatively regulated by RANKL.