Epigallocatechin-3-gallate Inhibits the Expression of Adhesion Molecules by Blocking Nuclear Factor Kappa B Signaling in Intestinal Epithelial Cells

BACKGROUND/AIMS: Epigallocatechin-3-gallate (EGCG) is the main polyphenol in green tea and has anti-inflammatory and anti-oxidative effects. The aim of this study was to determine the impact of EGCG on the expression of adhesion molecules and lipopolysaccharide (LPS)-induced nuclear factor-kappa B (NF-kappaB) signaling in rat intestinal epithelial (RIE) cells. METHODS: The effect of EGCG on LPS-induced NF-kappaB signaling and expression of intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 was examined by reverse transcription polymerase chain reaction, western blotting, immunofluorescence and electrophoretic mobility shift assay. RESULTS: LPS-induced expression of ICAM-1 and VCAM-1 mRNA was inhibited by EGCG treatment in RIE cells. LPS-induced inhibitor of kappa B alpha degradation and NF-kappaB nuclear translocation were blocked by EGCG in RIE cells. EGCG blocked LPS-induced NF-kappaB DNA-binding activity in RIE cells. The pharmacological NF-kappaB inhibitor Bay11-7082 suppressed the LPS-induced expression of ICAM-1 and VCAM-1 mRNA in RIE cells. CONCLUSIONS: These results indicate that EGCG inhibits LPS-induced ICAM-1 and VCAM-1 expression by blocking NF-kappaB signaling in intestinal epithelial cells.

Epigallocatechin-3-gallate Inhibits LPS-Induced NF-kappaB and MAPK Signaling Pathways in Bone Marrow-Derived Macrophages

BACKGROUND/AIMS: Epigallocatechin-3-gallate (EGCG), the primary catechin in green tea, has anti-inflammatory and anti-oxidative properties. The aim of the current study was to characterize the impact of EGCG on lipopolysaccharide (LPS)-induced innate signaling in bone marrow-derived macrophages (BMMs) isolated from ICR mice. METHODS: The effect of EGCG on LPS-induced pro-inflammatory gene expression and nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein kinase (MAPK) signaling was examined using reverse transcription-polymerase chain reaction, Western blotting, immunofluorescence, and the electrophoretic mobility shift assay. RESULTS: EGCG inhibited accumulation of LPS-induced IL-12p40, IL-6, MCP-1, ICAM-1, and VCAM-1 mRNA in BMMs. EGCG blocked LPS-induced IkappaBalpha degradation and RelA nuclear translocation. EGCG blocked the DNA-binding activity of NF-kappaB. LPS-induced phosphorylation of ERK1/2, JNK, and p38 was inhibited by EGCG. U0126 (an inhibitor of MEK-1/2) suppressed the LPS-induced IL-12p40, IL-6, MCP-1, ICAM-1, and VCAM-1 mRNA accumulation in BMMs. CONCLUSIONS: These results indicate that EGCG may prevent LPS-induced pro-inflammatory gene expression through blocking NF-kappaB and MAPK signaling pathways in BMMs.

Morphological Changes in Vasopressin-immunoreactive Neurons in the Hypothalamus of the Aged Rats / 체질인류학회지

The role of neuropeptides in the central nervous system (CNS) has received increasing attention. Numerous peptide molecules are found in the mammalian CNS and many of them are thought to act as either neurotransmitters or neuromodulators. The neuropeptides found in high concentration in the hypothalamus include vasopressin (VP), vasoactive intestinal polypeptide, somatostatin, and oxytocin. The main approches to assess the involvement of neuropeptides can be focused on functions affecting the aging of the brain. Morphological aging of the CNS has been characterized by degenerative changes of fiber connections and cell loss, although degeneration does not always occur to the same extent throughout various parts of the brain and, moreover, varies for different cell types. Despite of many studies in VP containing neurons , there exist discrepancies in results about the changes of aged rat brain. The aim of the present study is, therefore, to investigative possible changes in the number and morphology of VPimmunoreactive neurons with aging in each area of the hypothalmus of the aged rats. As a result, the number of VP-immunoreactive neurons was decreased in hypothalamus nucleus of aged group. Especially, in VP-immunoreactive neurons of hypothalamus, the size of neuronal cell body and nuclei in aged group is larger than in young group and the fiber density of immunoreactivity neurons of median eminance (ME) in aged group is stronger than in young group. But, the total number of VP-immunoreactive neurons in the suprachiasmatic nucleus (SCN) of the aged group is larger than in the young group. These studies indicate the involvement of VP-immunoreactive neurons in aging process of hypothalamus, and aging process may affect the synthesis of VP in the neurons of hypothalamic nuclei. Whereas, in VP expression, aging process induces an enlargement of the cell size of surviving neurons to compensate.

Effects of the Human-Mesenchymal Stem Cells Transplantation on Spinal Cord Injury of the Rats / 대한해부학회지

Two sources of adult stem cells that have aroused great interest are human bone marrow-derived mesenchymal stem cells (hMSCs) and human umbilical cord blood cells. hMSCs have been reported to maintain their ability to differentiate into neuronal lineage cells in the central nervous system. Therefore, transplantation of hMSCs represents an attractive new form of cellular therapy for clinical application in spinal cord injury (SCI). The aim of this study was to investigate how transplanted hMSCs from the venous circulation moved into a target zone of compression injury in the spinal cord of rats, and if they ameliorated the behavioral impairments associated with SCI. SCI in rats was induced by compressing the spinal cord for 30 s with an aneurysm clip. hMSCs labeled with cholera toxin subunit B conjugated to fluorescein isothiocyanate (CTX B-FITC) were injected intravenously through the tail vein or directly on the SCI site using a 27-g needle. Suspensions of hMSCs collected from adult humans were delivered at concentrations (1x10(6)cells/200 microliter) in 1 or 5 d after experimental SCI. After transplantation of hMSCs, the SCI regions displayed some endogenous background fluorescence, but CTX B-FITC-labeled hMSCs were clearly identifiable. They were observed in injured but not in intact areas; they were usually round or slightly elongated with a prominent nucleus. Only a few hMSCs were found in the spinal cord in each case but there were more cells in the rats injected at day one than at day five. This study confirmed that these were indeed transplanted hMSCs using antisera recognizing human-specific nuclei or human-specific mitochondria. Double immunofluorescence analysis showed the production of some neuronal and glial cell markers in the SCI lesions. Behavioral test scores of SCI rats treated with hMSCs at day one were significantly better than those for rats treated at day five and for the untreated SCI group. Thus, hMSCs appear to be beneficial in reversing the behavioral effects of SCI in this rat model, even when infused one day after injury. They might be a viable source of stem cells for the treatment of human neurological disorders.

Effect of Maternal Thyroxine Treatment on the Postnatal Development of Brain-derived Neurotrophic Factor-containing Neuron in the Brain of Pups of Alcohol Abused Mother / 대한해부학회지

Maternal alcohol abuse is thought to be the common cause of mental retardation. Especially, continuous alcohol consumption during critical period of brain development induce fetal alcohol effects. In this study, the authors investigated the effects of maternal alcohol drinking on the postnatal changes of BDNF contents and patterns of BDNF-containing neuron in neonatal rat brain, and, the influence of maternal thyroxine treatment on the brain of pups of alcohol abused mother. Pregnant rats were divided into three groups. Alcohol-fed group (n=4) received 35 calories of liquid alcohol diet daily from gestation day 6; control pair-fed group (n=4) was fed a liquid diet in dextrin replaced alcohol isocalorically; alcohol+T4 group (n=4) received 35 calories liquid alcohol diet and exogenous thyroxine (5 microgram/kg/day) subcutaneously. The amount of BDNF was significantly higher in the alcohol+T4 group as compared to the alcohol group at P7, P14 and P21, especially, alcohol+T4-exposed pups showed a significant increase of BDNF at P7. The decrease in BDNF was found in alcohol group compared to control pair-fed group at all ages. In alcohol+T4 group, BDNF-containing Purkinje cells exhibited mature pattern and monolayer arrangement at P14. Alcohol+T4 group showed mature pattern and numerical increase of BDNF-containing cells in cerebral cortex, hypothalamus and hippocampus at P7. The BDNF immunoreactivity of hippocampus continued to show prominent configuration in alcohol+T4 group at P28. These results indicate that the increase of the BDNF-containing neurons and BDNF amount in pups of thyroxinesupplemented alcohol-exposed dams as compared to control pair-fed and alcohol-exposed pups at P7, presumably suggest the early postnatal growth stimulatory effect of the exogenously supplemented thyroxine. Therefore, the increase of BDNF synthesis caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects, one of the ill effects as a result of the dysthyroid state following maternal alcohol abuse.

Effect of Chronic Alcohol Intake on Vasopressin and Oxytocin-containing Neurons in the Paraventricular and Supraoptic Nucleus of the Rat Hypothalamus / 체질인류학회지

Chronic alcohol intake can profoundly modify the neuronal activity and the morphologic structure of hypothalamic nucleus in the rat brain. The aim of the present study is to observe the effects of chronic alcohol intake on expression of vasopressin and oxytocin in the paraventricular and supraoptic nucleus in the rat hypothalamus. Experimental rats (n=14) were divided into control group and chronic alcohol group. Chronic alcohol group was induced via daily liquid alcohol intake for 6 months beginning at 8 weeks of age. As a result, the number of vasopressin and oxytocin-containing neurons was decreased in the paraventricular and supraoptic nucleus in chronic alcohol group. Especially, the number of vasopressin-containing neurons of chronic alcohol group was significantly decreased in the paraventricular nucleus. Chronic alcohol intake produced significant changes in the volume of the cell bodies and their nucleus in neurons of the paraventricular and supraoptic nucleus. Particularly, the size of nucleus of vasopressin-containing neurons in chronic alcohol group was larger than in control group. These results show that chronic alcohol intake may affect the synthesis of vasopressin and oxytocin in the neurons of hypothalamic nuclei. Whereas, chronic alcohol intake induces an enlargement of the cell size of surviving neuron to compensate.

Effect of Maternal Thyroxine Treatment on the Postnatal Development of Brain-derived Neurotrophic Factor-containing Purkinje Cell in Fetal Alcohol Effects in the Rat Cerebellum / 체질인류학회지

Maternal alcohol abuse is thought to be the common cause of mental retardation. Even moderate maternal alcohol consumption may produce fetal alcohol effects with behavioral and learning difficulties, if the drinking is associated with malnutrition. Especially, continuous alcohol consumption during critical period of brain development is very likely to produce fetal alcohol effects. The aims of this study are to investigate whether exogenous thyroxine treatment to alcohol -fed dams may ameliorate the detrimental effects of alcohol on the postnatal development of BDNF -containing Purkinje cell of the cerebellar cortex of the offspring. The morphological features of the growth and maturation were observed at 0, 7, 14, 21, 28 postnatal days via immunohistochemistry. In addition, electron microscopic finding of BDNF -containing Purkinje cell at P14 was also examined. Time -pregnant rats were divided into three groups. Alcohol -fed group received 35 calories of liquid alcohol diet daily from gestation day 6; control pair -fed group was fed a liquid diet in which dextrin replaced alcohol isocalorically; alcohol +/-T4 group received 35 calories liquid alcohol diet and exogenous thyroxine subcutaneously. As a result, a similar developmental pattern of BDNF -immunoreactive Purkinje cells was observed in control pair - fed and alcohol+/-T4 group on and after P14. These cells of alcohol -fed group showed immature features. Single -layer arrangement of these cells in alcohol -fed group was not completely achieved throughout postnatal life. Electron microscopic observations of BDNF -immunoreactive Purkinje cells at P14 revealed large nucleus, small cytoplasm, small amount of ribosomal collection and rudimentary cytoplasmic organelles in alcohol -fed group. The morphology of BDNF -immunoreactive Purkinje cell in alcohol +/-T4 group was similar to that in control pair -fed group. It was characterized by numerous short segments of rough endoplasmic reticulum, many of which showed a tendency of parallel alignment that suggested an attempt at Nissl body configuration. The cytology of Golgi complexes was also found within the cytoplasm in perinuclear location. Those observed differences of postnatal maturation patterns between alcohol -fed and alcohol +/-T4 group may indicate the beneficial effects on the postnatal development of BDNF -containing Purkinje cells in cerebellar cortex in the pups of thyroxine -treated alcohol -exposed dams. These results suggest that the increase of BDNF synthesis during early postnatal life caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects as a result of the dysthyroid state following maternal alcohol abuse.

Postnatal Development of Brain Natriuretic Peptide-immunoreactive Neuron in the Hypothalamus of the Rat / 체질인류학회지

Brain natriuretic peptide (BNP) is a neuropeptide, isolated from porcine brain that is homologous with atriopeptin. Magnocellular neurosecretory cells located in the paraventricular nucleus and supraoptic nucleus synthesize and secrete neurohormones. The purpose of this study was to investigate distribution of BNP immunoreactivity throughout the rat hypothalamus from the day of birth to 30 days and adult using immunoperoxidase and immunofluorescent staining. The first BNP immunoreactive neurons appeared in the paraventricular and supraoptic nucleus at P10. In adult, BNP immunoreactivity was widely distributed throughout regions of the hypothalamus including dorsomedial hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus and internal layer of median eminence. The intensity of BNP immunoreactivity was weak in almost all hypothalamic nuclei except the paraventricular and supraoptic nuclei. BNP immunoreactivity was first observed in the lateral hypothalamic area at P15. In retrochiasmatic supraoptic nucleus, BNP immunoreactivity was first observed at P20 and remarkably distributed in adult. In the present study, distinct localization of BNP immunoreactivity was in the hypothalamic cell bodies and fibers. Although the role of BNP in the brain is yet to be determined, these results indicate that BNP in the neurons of hypothalamus play important role in the regulation of a variety of neurosecretory functions as a neuromodulator during postnatal development of the hypothalamus.

Postnatal Development of Brain Natriuretic Peptide-immunoreactive Neuron in the Hypothalamus of the Rat / 체질인류학회지

Brain natriuretic peptide (BNP) is a neuropeptide, isolated from porcine brain that is homologous with atriopeptin. Magnocellular neurosecretory cells located in the paraventricular nucleus and supraoptic nucleus synthesize and secrete neurohormones. The purpose of this study was to investigate distribution of BNP immunoreactivity throughout the rat hypothalamus from the day of birth to 30 days and adult using immunoperoxidase and immunofluorescent staining. The first BNP immunoreactive neurons appeared in the paraventricular and supraoptic nucleus at P10. In adult, BNP immunoreactivity was widely distributed throughout regions of the hypothalamus including dorsomedial hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus and internal layer of median eminence. The intensity of BNP immunoreactivity was weak in almost all hypothalamic nuclei except the paraventricular and supraoptic nuclei. BNP immunoreactivity was first observed in the lateral hypothalamic area at P15. In retrochiasmatic supraoptic nucleus, BNP immunoreactivity was first observed at P20 and remarkably distributed in adult. In the present study, distinct localization of BNP immunoreactivity was in the hypothalamic cell bodies and fibers. Although the role of BNP in the brain is yet to be determined, these results indicate that BNP in the neurons of hypothalamus play important role in the regulation of a variety of neurosecretory functions as a neuromodulator during postnatal development of the hypothalamus.

Postnatal Development of Transforming Growth factor-alpha-immunoreactive Neurons in the Diencephalon of the Rat / 체질인류학회지

Transforming growth factor-alpha (TGF-alpha) immunoreactivity during postnatal development was examined in the rat diencephalon using immunohistochemistry. The time of appearance and localization of TGF-alpha immunoreactivity was slightly different in many areas of diencephalon during postnatal development. At birth, TGF-alpha immunoreactivity was mainly evident in thalamic medial, median and parafascicular thalamic nucleus of intralaminar nuclei. In addition, TGF-alpha immunoreactivity was clearly evident at the first postnatal week in most hypothalamic nuclei. Therefore, TGF-alpha immunoreactivity was found at postnatal days 7 in most diencephalic nuclei excepting the vental posterior thalamic nuclei, metathalamus and epithalamus. The quantitative increase of number was first apparent in the midline structures of thalamus in the first postnatal week. And then TGF-alpha-immunoreactive cells progressively increased throughout diendephalon by postnatal days 15. Adult patterns were reached at postnatal days 20. These results indicate that TGF-alpha-immunoreactive cells were first appeared in thalamic midline structures, increased progressively in the first two postnatal weeks, and followed mediolateral gradient. In addition to maturation of TGF-alpha-immunoreactive cells requires a relatively prolonged period of time to achieve an adult configuration. Also, the early appearance of TGF-alpha immunoreactivity in most diencephalic nuclei may be related to the early appearance of EGFR immunorecativity in many other brain regions. Taken together, these findings suggest that TGF-alpha immunoreactivity correlated with the appearance of the related functional activity in the different regions of diencephalon.

Postnatal Development of Transforming-growth Factor-alpha and Epidermal Growth Factor Receptor-immunoreactive Neurons in the Cerebral Cortex of the Rat / 대한해부학회지

Transforming growth factor-alpha (TGF-alpha ) and epidermal growth factor receptor (EGFR) immunoreactivities were examined in the cerebral cortex of the rat during postnatal development. TGF-alpha -immunoreactive cells were found at birth in all cortical layers except the molecular layer. TGF-alpha -immunoreactive cells were most abundant in the parietal cortex at P20. The intensity and number of the TGF-alpha -immunoreactive cells increased at postnatal days 15 or 20 (P15 - P20). Mature patterns of TGF-alpha -immunoreactive cells were achieved at P20. EGFR -immunoreactive cells appeared only in dorsal endopiriform cortex at P0. The first EGFR -immunoreactive cells were observed in the neocortex at P3. These cells were most abundant in the parietal cortex at P90. In adult, the most prominent EGFR immunoreactivity occured in layer IV, V and VI. These cells were numerous in the frontal and parietal cortex, diminishing laterally towards the insular cortex. Adult patterns were reached on and after P10. The time of appearance and localization of EGFR immunoreactivity correlated with functional activity in the different cortical areas. No clear labelling of glial cells with TGF-alpha and EGFR antibodies was found. TGF -alpha and EGFR immunoreactivity was observed in the majority of neurons in the postnatal developing and adult cerebral cortex of the rat. Also double -immunohistochemistry with antibodies to TGF-alpha and EGFR showed co-localization of TGF -alpha and EGFR in neurons of the cerebral cortex. Co-localization of TGF-alpha and EGFR was first detectable in most cortices at P3. By P10, these neurons showed immature neuronal features. The present results showing TGF -alpha and EGFR immunoreactivity is widely distributed in the postnatal developing (except P0) and adult cerebral cortex, mainly localized in neurons. And TGF-alpha and EGFR co-localize in most neurons, thus indicating that most EGFR -containing cells are TGF-alpha -synthesizing cells. In addition to difference of time of appearance and mature neuronal pattern suggest that TGF-alpha has the capacity of activating the EGFR in the normal postnatal developing cerebral cortex, therefore, TGF-alpha and EGFR may interact within cortical neurons through many different mechanism according to postnatal age.