• The Korea Journal of Herbology
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• v.26 no.4
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• pp.83-88
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• 2011

Objectives : The aim of this study was to investigate the asthma-suppressive and immuno-regulatory effect of Notopterygii Rhizoma(NR) extract on OVA(ovalbumin)-induced asthma in mice. Methods : C57BL/6 mice out of all the experimental groups, except the Normal group and the NRI group, were sensitized and challenged with OVA. C57BL/6 mice were exposed to OVA three times a week for 12 weeks and analyzed by flow cytometer, ELISA, H&E stain. Results : The concentrations of IL-4, IL-5, IL-13, IgE in serum of the OVA-NRII group decreased significantly compared with those of the OVA-Control group. The number of $Gr-1^+/CD11b^+$, $CCR3^+$, $CD3^+/CD19^+$, $CD3e^+/CD69^+$cells in the OVA-NRI group decreased significantly compared with those of the OVA-Control group. The collagen accumulation in the lung sections of the OVA-NRII group decredased signi- ficantly compared with that of the OVA-Control group. Conclusions : These results suggest that Notopterygii Rhizoma(NR) would be a effective candidate for herbal-originated anti-asthmatic drug. However, this drug should be further studied for characterization of the accurate action and underlying mechanism using variant disease model in the future.

• International Journal of Oral Biology
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• v.44 no.3
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• pp.81-88
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• 2019

Trifolium pratense leaves (red clover) has been used in Oriental and European folk medicine for the treatment of whooping cough, asthma, and eczema, and is now being used to treat and alleviate the symptoms, such as hot flushes, cardiovascular health effects that occur in postmenopausal women. However, relatively little scientific data is available on the physiological activity of this plant. Therefore, in this study, we investigated the anti-cancer activity of T. pratense leaves using methanol extract of T. pratense leaves (MeTP) on human FaDu hypopharyngeal squamous carcinoma cells. MeTP inhibited the viability of FaDu cells by inducing apoptosis through the cleavage of procaspase-3, -7, and -9 and poly (adenosine diphosphate ribose-ribose) polymerase (PARP), downregulation of Bcl-2, and upregulation of Bax, as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Live & dead assay, 4'6-diamidino-2-phenylindole stain, fluorescence-activated cell sorting analysis, and Western blot analysis. In addition, colony formation was slightly inhibited when FaDu cells were treated with a non-cytotoxic concentration (0.125 mg/mL) of MeTP and almost completely inhibited when cells were treated with 0.25 mg/mL MeTP. Collectively, these results indicate that MeTP induced cell apoptosis via caspase- and mitochondrial-dependent apoptotic pathways, and inhibited colony formation of cancer cells in FaDu human hypopharyngeal squamous carcinoma cells. These findings suggest MeTP should be considered for clinical development as a chemotherapeutic option in oral cancer.

• The Korea Journal of Herbology
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• v.29 no.5
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• pp.75-81
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• 2014

Objectives : This study was performed to investigate the effect of root extract of Pueraria thunbergiana Bentham (Puerariae Radix, PR) in diabetic mice as similar as emaciation-thirst disease in Oriental medicine. Methods : C57BL/6 mice were fed high fat (HF) and high sucrose (HS) for 8 weeks, and then administrated with 90 mg/kg body weight (bw) of streptozotocin (STZ) for induction of diabetes which is similar to the middle emaciation stage. After 5 days, blood glucose levels were measured, and selected the mice with ranges above $250mg/d{\ell}$. PR water extract was administrated orally once a day for 4 weeks with high fat and high sucrose. The levels of glucose, insulin, total cholesterol, triglyceride, ${\gamma}glutamyl$ transpeptidase (${\gamma}GTP$), glutamic oxaloacetic transaminase (GOT) and glutamate pyruvate transaminase (GPT) were analysed in the serum. Also, observed their histological changes by hematoxylin and eosin (H&E) of different organs, lung, heart, pancreas, stomach, liver, and kidney. Results : PR extract significantly decreased the levels of serum glucose and insulin in diabetic mice. PR extract significantly increased the levels of triglyceride, total cholesterol, GOT and GPT in diabetic mice. In H&E stain, PR extract inhibited the histopathological changes of lung (as a channel of the upper emaciation stage in the channel-tropism theory), pancreas (as a channel of the middle emaciation stage) and kidney (as a channel of the lower emaciation stage) in diabetic damage. Conclusions : PR extract has an anti-diabetic effect in HF/HS and low-dose STZ-induced diabetic mice. This result suggests that PR follows the channel-tropism theory in the emaciation-thirst disease through the protection of lung, pancreas and kidney.

• The Korea Journal of Herbology
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• v.29 no.3
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• pp.71-78
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• 2014

Objectives : This study evaluates the neuroprotective effects of Sopung-tang, a mixture of Notopterygii Rhizoma, Saposhnikoviae Radix, Angelicae Gigantis Radix, Cnidii Rhizoma, Hoelen, Aurantii Nobilis Pericarpium, Pinelliae Tuber, Linderae Radix, Angelicae Dahuricae Radix, Cyperi Rhizoma, Cinnamomi Ramulus, Asari Radix, Glycyrrhizae Radix on the cerebral infarct combined with hyperlipidemia. Method : The hyperlipidemia was induced by the beef tallow 30% diet for 14 days on Sprague-Dawley rats. The cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) for 2 hours with intraluminal thread method. Then the water extract of Sopung-tang was administered a day for 5 days at 3 hours after the cerebral infarct by MCAO. Effect of Sopung-tang was evaluated with the infarct volume and edema percentage by a TTC-stained brain section, and the expressions of Bax and Bcl-2 in the brain tissue by a immunohistochemical stain method. Results : Sopung-tang reduced the infarct size partly in a TTC-stained brain section of the hyperlipidemic MCAO rats. Sopung-tang reduced the infarct volume of the hyperlipidemic MCAO rats significantly. Sopung-tang reduced the edema percentage of the hyperlipidemic MCAO rats, but not significant statistically. Sopung-tang suppressed the Bax expressions in the cerebral penumbra and caudate putamen of the hyperlipidemic MCAO rats significantly. Sopung-tang upregulated the Bcl-2 expression in the caudate putamen of the hyperlipidemic MCAO rats. Conclusion : These results suggest that Sopung-tang plays an anti-apoptotic neuroprotective effect through the suppression of Bax and up-regulation of Bacl-2 expressions in the brain tissues.

• Tuberculosis and Respiratory Diseases
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• v.43 no.3
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• pp.420-433
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• 1996

Background : There have been many debates about the effects of nitric oxide on the neurogenic inflammation. The role of nitric oxide in the neurogenic inflammation of airways will be required a better understanding of the localization and types of nitirc oxide synthase(NOS) activity in the neurogenic inflammation of airways. Method : To investigate the role of nitric oxide in airway neurogenic inflammation, 1) the effects of neurokinin receptor antagonist (FK224) and nitric oxide synthase inhibitor, $N^{\omega}$-nitro-L-arginine (L-NNA) on plasma extravastion were evaluated in four groups of Sprague-Dawley rats ; sham operation group(sham NANC group), electrical vagal stimulation group(NANC2 group), intravenous pretreatment groups with FK224 (1mg/kg ; FK224 group), and L-NNA(1mg/kg ; L-NNA group) 15 minutes before vagal NANC stimulation. 2) NOS activity in trachea with neurogenic inflammation was localized by immunohistochemical stain. Immunohistochemical stain was performed by antibodies specific for inflammatory cells(iNOS), brain(bNOS), and endothelium (eNOS) on trachea obtained from sham NANC, NANC2, and FK224 groups. Results : The results are that plasma extravsation in neurogenic inflammation of rat airways was inhibited by FK224, but enhanced by L-NNA pretreatment(P<0.05). There was significantly increased infiltration of inflammatory cells in subepithelium of neurogenic inflammatory trachea, but the reduction of subepithelial infiltration of inflammatory cells was observed after pretreatment with FK224(P<0.05). Immunostaining with anti-iNOS antibody showed strong reactivity only in infiltrated inflammatory cells in neurogenic rat trachea, and these iNOS reactivity was reduced by pretreatment with FK224. bNOS immunoreactivity was significantly increased only in the nerves both of neurogenic inflammatory and FK224 pretreated trachea compared with sham NANC trachea(p<0.05). eNOS immunoreactivity was not significant change in endothelium in neurogenic inflammation of rat trachea. Conclusion : These results suggest that nitric oxide released from iNOS in infiltrated inflammatory cells has main role in neurogenic inflammation of rat trachea. The presence of bNOS immunoreactivity in the nerves indicates that nitric oxide may be released from the nerves in rat trachea with neurogenic inflammation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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• pp.1055-1060
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• 2004

Apoptosis is the ability of cells to self-destruct by the activation of an intrinsic cellular suicide program when the cells are no longer needed or when they are seriously damaged. Morphologically, apoptosis is characterized by the appearance of membrane blebbing, cell shrinkage, chromatin condensation, DNA cleavage, and the fragmentation of the cell membrane-bound apoptotic bodies. Siegesbeckia glabrescens Makino (Siegesbeckiae Herba, SG) has been widely used as treatments for arthritis, and fever, as well as detoxification properties. The present studies were undertaken to evaluate if SG has an anti-apoptotic property. Cell viability was measured by XTT and tryphan blue stain. Morphological characteristic of human aortic smooth muscle cells(HASMC) were visualized with a phase-contrast microscope. SG significantly reduced HASMC, but not human umbilical vein endothelial cell(HUVEC), viability in a dose-dependent manner. Confluent untreated cells at 24hrs showed normal morphology, flat with a uniform polygonal shape. SG-treated cells (0.5㎎/㎖) at 24hrs showed apoptotic morphology. Cells became irregular with elongated lamellipodia, and exhibited condensed chromatin in nuclei with occasional endoucleation. There was an increase in the number of apoptotic cells rounding-up and being detached from the substrate. TUNEL staining of SG-treated cells showed dark-brown stains in nuclei and cytosol. Caspases are central components of the machinery responsible for apoptosis and are generally divided into two categories; the initiator caspases, which include caspases-2,-8,-9, and -10, and the effector caspases, which include caspases-3,-6, and -7. SG decreased anti-caspase-3 protein expression, which means activation of caspases-3 activity. It has been reported that there is a link between NO formation and apoptosis. NO production was accelerated by SG treatment in HASMC. L-NNA, NOS inhibitor, inhibited SG-induced apoptosis. These results, therefore, indicated that both caspases-3 and NO production are involved in apoptosis in smooth muscle cells. According to these results, SG may have a potential effect in the treatment of hypertensive atherosclerosis.

• Journal of Korean Medicine Rehabilitation
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• v.28 no.1
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• pp.19-31
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• 2018

Objectives The object of this study was to investigate the antioxidative and antiinflammatory effects of Jinmu-tang extract (JMT) on the Monosodium iodoacetate (MIA)-induced rat osteoarthritis. Methods To investigate the antioxidant capacities of JMT, we measured the total polyphenol and flavonoid, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-Azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity. To evaluate the antioxidative and antiinflammatory effects of JMT, the rats were divided into 5 groups (n=8). Normal group was not induced by MIA and treated at all (N), control group was induced by MIA and not treated at all (Con), positive control group was induced by MIA and orally administered indomethacin 5 mg/kg (Indo) and experimental groups were induced by MIA and orally administered JMT 100 mg/kg (JMT100) and JMT 200 mg/kg (JMT200) for 4 weeks. The changes of anti-type II collagen antibody in serum, heme oxygenase-1 (HO-1), phosphorylated inhibitor of ${\kappa}B{\alpha}$ ($p-I{\kappa}B{\alpha}$), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and tumor necrosis factor alpha ($TNF-{\alpha}$) in knee joint tissue and histopathological observation (Hematoxylin & Eosin and Safranin-O stain) were measured. Results Total polyphenol and flavonoid levels of JMT were $26.90{\pm}0.33mg/g$ and $6.02{\pm}0.34mg/g$. $IC_{50}$ of L-ascorbic acid and JMT of DPPH radical scavenging activity were $1.35{\pm}0.07{\mu}g/ml$ and $52.95{\pm}0.97{\mu}g/ml$. $IC_{50}$ of L-ascorbic acid and JMT of ABTS radical scavenging activity were $3.18{\pm}0.02{\mu}g/ml$ and $91.49{\pm}1.74{\mu}g/ml$. In serum, the anti-type II collagen antibody levels of JMT100 and JMT200 groups were decreased significantly. In knee joint tissue, the HO-1 level of JMT200 was increased significantly. The $p-I{\kappa}B{\alpha}$ and $TNF-{\alpha}$ levels of JMT200 were decreased significantly. The COX-2 and iNOS levels of JMT groups were decreased significantly. In histopathological observation, in comparison with Con, synovial tissue, cartilage and proteoglycan of JMT100 and JMT200 were well preserved. Conclusions According to the results, It is considered that JMT has antioxidant and antiinflammatory effects for MIA-induced rat osteoarthritis, so it could be applied to osteoarthritis treatment.

• Applied Microscopy
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• v.31 no.1
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• pp.1-8
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• 2001

The morphological features of neuronal dendritic spines are changed their shapes, sizes and density in response to physiological or pathological conditions . Therefore, exact analysis of spines warrants understanding of neuronal function. The size of the spine is at the borderline of resolution with light microscopy. High voltage electron microscopy Provide excellent resolution of the spines with proper stain techniques thanks to its higher resolution and penetration power. We evaluated more effective staining method for observing dendritic spines after labeling Purkinje cells with anti-calbindin 28 kD immunohistochemistry or Golgi staining methods. 4 fm thickness sections were observed with high voltage electron microscopy and some morphometric analyses were performed. Both Golgi staining and immunohistochemistry revealed the detail structures of the Purkinje cell such as soma, dendrites, and dendritic spines. High voltage electron micrographs with Golgi staining provide more precise morphology and are easy to measure. Average density of spine is $24.5{\pm}3.6/10{\mu}m$ and its length is $1.12{\pm}0.22{\mu}m$. For quantitative analysis of the spines, high voltage electron, micrographs with Golgi staining are more effective. This preliminary result is expected to be useful for further study of spine plasticity in various conditions.

• Herbal Formula Science
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• v.24 no.3
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• pp.163-174
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• 2016

Objectives : Rheum undulatum Linne and Glycyrriza uralensis Fischer are widely used herbal medicine. In this study, anti-oxidant and liver protective effects of R. undunlatum extract (RUE) and G. uralensis extract (GUE) were investigated in HepG2 cells, respectively. Oxidative stress and liver fibrosis were induced by arachidonic acid (AA) and iron, and CCl₄.Methods : MTT assay was assessed for cell viability, and immunoblotting analysis was performed to detect expression of apoptosis related proteins. In addition, reactive oxygen species (ROS) and mitochondrial dysfunction were measured. In vivo, BALB/c mouse were orally administrated with the aqueous extract of 10 mg/kg RUE and 100 mg/kg GUE for 3 days and then, injected with CCl₄ 0.5 ml/kg body weight to induce acute liver damage. Serum ALT level was measured, and histological change was observed in Harris's hematoxylin and eosin stainResults : RUE and GUE pre-treatment increased relative cell viability in concentration dependent manner and altered the expression levels of apoptosis-related proteins such as procaspase 3, PARP and Bcl-xL. RUE and GUE also inhibited the mitochondrial dysfunction and excessive reactive oxygen species (ROS) production induced by AA and iron. In addition, RUE and GUE activated liver kinase B1 (LKB1), by increasing phosphorylation. Moreover, RUE and GUE treatment decreased liver injuries induced by CCl₄, as evidenced by decreases in histological liver damage as well as serum alanine amino transferase (ALT) level.Conclusions : These data suggest that RUE and GUE has anti-oxidant and liver protective effects against AA and iron-induced oxidative stress and CCl₄-induced liver injury.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.24 no.4
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• pp.377-383
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• 2021

Purpose: We investigated the association of effector memory (EM) CD8⁺ T cell and CD4⁺ T cell immunity with metabolic syndrome (MS). Methods: Surface and intracellular staining of peripheral blood mononuclear cells was performed. Anti-interleukin-7 receptor-alpha (IL-7Rα) and CX3CR1 antibodies were used to stain the subsets of EM CD8⁺ T cells, while anti-interferon-gamma (IFN-γ), interleukin-17 (IL-17), and forkhead box P3 (FOXP3) antibodies were used for CD4⁺ T cell subsets. Results: Of the 47 obese children, 11 were female. Children with MS had significantly higher levels of serum insulin (34.8±13.8 vs. 16.4±6.3 µU/mL, p<0.001) and homeostasis model assessment of insulin resistance (8.9±4.1 vs. 3.9±1.5, p<0.001) than children without MS. Children with MS revealed significantly higher frequencies of IL-7Rα^low CD8⁺ T cells (60.1±19.1% vs. 48.4±11.5%, p=0.047) and IL-7Rα^lowCX3CR1⁺ CD8⁺ T cells (53.8±20.1% vs. 41.5±11.9%, p=0.036) than children without MS. As the serum triglyceride levels increased, the frequency of IL-7Rα^lowCX3CR1⁺ and IL-7Rα^highCX3CR1^- CD8⁺ T cells increased and decreased, respectively (r=0.335, p=0.014 and r=-0.350, p=0.010, respectively), in 47 children. However, no CD4⁺ T cell subset parameters were significantly different between children with and without MS. Conclusion: In obese children with MS, the changes in immunity due to changes in EM CD8⁺ T cells might be related to the morbidity of obesity.