• Journal of Animal Science and Technology
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• v.62 no.5
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• pp.595-604
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• 2020

This study investigated the effects of dietary rumen-protected L-tryptophan (TRP) supplementation (43.4 mg of L-tryptophan kg^-1 body weigt [BW]) for 65 days in Hanwoo steers on muscle development related to gene expressions and adipose tissue catabolism and fatty acid transportation in longissimus dorsi muscles. Eight Hanwoo steers (initial BW = 424.6 kg [SD 42.3]; 477 days old [SD 4.8]) were randomly allocated to two groups (n = 4) of control and treatment and were supplied with total mixed ration (TMR). The treatment group was fed with 15 g of rumen-protected TRP (0.1% of TMR as-fed basis equal to 43.4 mg of TRP kg^-1 BW) once a day at 0800 h as top-dressed to TMR. Blood samples were collected 3 times, at 0, 5, and 10 weeks of the experiment, for assessment of hematological and biochemical parameters. For gene study, the longissimus dorsi muscle samples (12 to 13 ribs, approximately 2 g) were collected from each individual by biopsy at end of the study (10 weeks). Growth performance parameters including final BW, average daily gain, and gain to feed ratio, were not different (p > 0.05) between the two groups. Hematological parameters including granulocyte, lymphocyte, monocyte, platelet, red blood cell, hematocrit, and white blood cell showed no difference (p > 0.05) between the two groups except for hemoglobin (p = 0.025), which was higher in the treatment than in the control group. Serum biochemical parameters including total protein, albumin, globulin, blood urea nitrogen, creatinine phosphokinase, glucose, nonesterified fatty acids, and triglyceride also showed no differences between the two groups (p > 0.05). Gene expression related to muscle development (Myogenic factor 6 [MYF6], myogenine [MyoG]), adipose tissue catabolism (lipoprotein lipase [LPL]), and fatty acid transformation indicator (fatty acid binding protein 4 [FABP4]) were increased in the treatment group compared to the control group (p < 0.05). Collectively, supplementation of TRP (65 days in this study) promotes muscle development and increases the ability of the animals to catabolize and transport fat in muscles due to an increase in expressions of MYF6, MyoG, FABP4, and LPL gene.

• Journal of Haehwa Medicine
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• v.24 no.2
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• pp.35-57
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• 2016

Objectives : The purpose of this study is to prove the effect and mechanism of Gamikyejakjimogawusul-tang(GKHA) herbal acupuncture on induced rheumatoid arthritis model of DBA/1 mice. Methods : We check effect of GKHA extract on the AST, ALT, Creatinine, BUN of serum and cell viability of GK extract in RAW 264.7 cells to test the stability of this study. In vitro, we measure total phenol contents, total flavonoid contents, DPPH free radical scavenging activity, ABTS cation radical scavenging activity of Gamikyejakjimogawusul-tang, effect of GK extract on ROS(Reactive Ooxygen Species) production to estimate a anti-oxidant capacity, and we also measure effect of GK extract on NO (Nitric Oxid), IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF production in RAW 264.7 cells to estimate a anti-inflammatory efficacy. In vivo, we compare a rheumatoid arthritis manifestation between control and experimental group and estimate a AI. Then we check effect of GKHA on the level of WBC, neutrophil, lympocyte, monocyte in the blood to see the effect of immune cells in blood. In addition we measure effect of GKHA on the level of hs-CRP, IgM, IgG, IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF in serum. We observe effects of GKHA on imaging of cartilage degeneration using micro CT-arthrography in paw hind. And we calculate effects of GKHA that reduced BV ratio, BS/BV ratio using 3D Micro-CT. Lastly we observe effects of GKHA histopathologic examination analysis. Results : 1. The toxicity on liver and kidney was disregardable and the cytotoxicity against RAW 264.7 cells was also disregardable. 1. Total phenol contents and total flavonoid contents in GK extract were in high level. 2. DPPH free radical scavenging activity and ABTS cation radical scavenging activity were increased according to concentration of GK extract 3. ROS production was significantly decreased in GK extract (at 10, $100{\mu}g/ml$). 4. NO, IL-6, TNF-${\alpha}$, MCP-1 production were significantly decreased in GK extract(at 10, $100{\mu}g/ml$). IL-17, GM-CSF production were significantly decreased in GK extract(at 1, 10, $100{\mu}g/ml$). IL-$1{\beta}$, IL-21 production were also decreased but there was no statistical significance. 5. 25x observation after H&E and M-T staining, infiltration of immune cells and subsidence of the cartilage and damage to the synovial cells were decreased. Conclusions : This study showed that GKHA extract had anti-oxidant capacity, anti-inflammatory efficacy. GKHA extract also had inhibiting effect on the process of rheumatoid arthritis and can protect joint and cartilage. So we expect that GKHA extract can be a meaningful treatment to rheumatoid arthritis patients.

• Journal of Life Science
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• v.30 no.3
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• pp.267-277
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• 2020

This study was conducted to assess the effect of highly concentrated onion intake on rodents. The experimental animals were divided two groups as follows; water administered group (CON) and Allium cepa administered group (ACE). The ACE group showed a slightly increases in the number of erythrocytes (RBC), hemoglobin (HGB), hematocrit (Hct) levels compared to the control group (p<0.05). Hemoglobin, monocyte, lymphocyte and neutrophil had no significant change (p>0.05) in ACE group and control group. The analysis of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) levels showed a significant decrease in ACE group compared to the control group (p<0.05). The blood glucose, total protein, HDL-cholesterol were slightly high in ACE group, while triglyceride, total cholesterol levels were lower in ACE group compared to the control group (p<0.05). The levels of cytokines (interluekin-1α (IL-1α), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), interferon-γ (INF-γ), interleukin-18 (IL-18), interleukin-2 (IL-2), granulocyte-macrophage colony-stimulating factor (GM-CSF)) involved in immunity and inflammation in liver tissue and blood have all been confirmed to be within normal range. These findings could be used as basic data to show that highly concentrated dietary onion extract is not toxic to hematological indicators and immune functions.

• Journal of Oriental Neuropsychiatry
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• v.28 no.3
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• pp.231-248
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• 2017

Objectives: This study evaluates anti-stress and sleep-inductive effects of Dongshingihyeolyangsubang (DSGYSB) on rats induced by chronic mild stress (CMS). Methods: Twenty-five healthy rats were randomly divided into five groups: normal, CMS (Control), DSGYSB50, DSGYSB100, and DSGYSBS200. All rats except the normal group were exposed to unpredictable stress conditions such as water deprivation, empty bottles, and forced treadmill. A week after starting the experiment, rats in DSGYSB50, DSGYSB100, and DSGYSB200 groups were fed orally with water once a day for two weeks. Then blood samples were taken from the rats for analysis of complete blood count, AST, ALT, and glucose. Noradrenaline, corticosterone, serotonin, GABA and melatonin were measured by ELISA kit. BDNF, CREB, TrkB and $TNF-{\alpha}$ were measured by RT-PCT. Results: In Noradrenaline contents, the DSGYSB200 group revealed significant decrease compared to the control group. In corticosterone contents, the DSGYSB200 group revealed significant decrease compared to the control group. In serotonin contents, the DSGYSB200 group revealed significant increase compared to the control group. In GABA contents, the DSGYSB50, DSGYSB100, and DSGYSB200 groups revealed significant increase compared to the control group. In the activity of BDNF, the DSGYSB50, DSGYSB100 and DSGYSB200 groups revealed significant increase compared to the control group. In the activity of CREB, the DSGYSB100 and DSGYSB200 groups revealed a significant decrease compared to the control group. In the activity of TrkB, the DSGYSB100 and DSGYSB200 groups revealed significant decrease compared to the control group. In the activity of $TNF-{\alpha}$, DSGYSB100 and DSGYSB200 groups revealed significant decrease compared to the control group. In glucose contents, the DSGYSB100 and DSGYSB200 groups revealed significant decrease compared to the control group. In the leukocyte changes, white blood cells, neutrophil, lymphocytes, and monocyte significantly increased in the DSGYSB100 and DSGYSB200 groups than the control group. In the erythrocyte changes, hemoglobin significantly increased in the DSGYSB200 group than the control group. Conclusions: Results suggest that Dongshingihyeolyangsubang has anti-stress and sleep-inductive effects on rats induced by CMS.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.4
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• pp.377-384
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• 2017

Activation of protein kinase C (PKC) is closely linked with endothelial dysfunction. However, the effect of $PKC{\beta}II$ on endothelial dysfunction has not been characterized in cultured endothelial cells. Here, using adenoviral $PKC{\beta}II$ gene transfer and pharmacological inhibitors, the role of $PKC{\beta}II$ on endothelial dysfucntion was investigated in cultured endothelial cells. Phorbol 12-myristate 13-acetate (PMA) increased reactive oxygen species (ROS), p66shc phosphorylation, intracellular adhesion molecule-1, and monocyte adhesion, which were inhibited by $PKC{\beta}i$ (10 nM), a selective inhibitor of $PKC{\beta}II$. PMA increased the phosphorylation of CREB and manganese superoxide dismutase (MnSOD), which were also inhibited by $PKC{\beta}i$. Gene silencing of CREB inhibited PMA-induced MnSOD expression, suggesting that CREB plays a key role in MnSOD expression. Gene silencing of $PKC{\beta}II$ inhibited PMA-induced mitochondrial ROS, MnSOD, and ICAM-1 expression. In contrast, overexpression of $PKC{\beta}II$ using adenoviral $PKC{\beta}II$ increased mitochondrial ROS, MnSOD, ICAM-1, and p66shc phosphorylation in cultured endothelial cells. Finally, $PKC{\beta}II$-induced ICAM-1 expression was inhibited by Mito-TEMPO, a mitochondrial ROS scavenger, suggesting the involvement of mitochondrial ROS in PKC-induced vascular inflammation. Taken together, the results suggest that $PKC{\beta}II$ plays an important role in PMA-induced endothelial dysfunction, and that the inhibition of $PKC{\beta}II$-dependent p66shc signaling acts as a therapeutic target for vascular inflammatory diseases.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.2
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• pp.128-134
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• 2017

Der p 1 and Der p 2 are essential allergens of house dust mite associated with the development of asthma. In the present study, we examined whether Der p 1 and Der p 2 induce a release of S100A8 and S100A9 in monocytes, which are involved in the regulatory mechanism of neutrophil apoptosis. We found that Der p 1 and Der p 2 significantly increased the secretion of S100A8 and S100A9 in normal monocytes. Moreover, S100A8 and S100A9 strongly suppressed the spontaneous apoptosis of normal and asthmatic neutrophils. The inhibitory effect of S100A9 was stronger than that of S100A8, and asthmatic neutrophils showed a higher inhibitory effect than normal neutrophils. S100A8 and S100A9 induced activation of Lyn, Akt, and ERK in a time-dependent manner. These findings elucidate the roles of Der p 1 and Der p 2 in the interaction between monocytes and neutrophils, as well as contributing to our knowledge of the pathogenesis of allergic diseases.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.33 no.2
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• pp.83-99
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• 2020

Objectives : Atopic dermatitis is a chronic inflammatory skin disease with frequent relapses. This study was to investigate the effects of Gammakdaejo-tang(GMD) in DNCB induced atopic dermatitis mice. Methods : The study was divided into five comparion groups. 2,4-dinitrochlorobenzene(DNCB) solution was applied to Nc/Nga mice to induce atopic dermatitis, followed by normal group, negative control group with distilled water, positive control group with Dexamethasione and GMD 200mg/kg or 400mg/kg. The control group was orally administered 200㎕ once daily for 4 weeks. Visual skin condition, Immunoglobulin E, Histamine, Cytokine, Immune cells, Tissue biomarkers were observed. Results : As a result of the dermatitis score evaluation, it was confirmed that the GMD-administered group improved symptoms compared to the negative control group. As a result of measuring IgE, the GMD-administered group significantly decreased compared to the negative control group. As a result of measuring Histamine, GMD group except 200mg/kg of GMD significantly decreased compared to negative control group. As a result of measuring cytokine, GMD 200mg/kg significantly reduced IL-1β, IL-6 and TNF-α compared to the negative control. 400mg/kg significantly reduced IL-1β, IL-4, IL-5, IL-6, IL-10, TNF-α and significantly increased IL-2, IFNγ. As a result of confirming the immune cells, all experimental groups showed no difference in basophil, GMD group significantly reduced monocyte and eosinophil compared to negative control group, and GMD 400mg/kg group significantly reduced white blood cell and neutrophil. And significantly increased lymphocytes. As a result of measuring the gene expression level, all GMD group significantly increased TGF-β1 compared with the negative control group, and filaggrin, VEGF and EGF were significantly increased in GMD 400mg/kg group. Epidermis, dermis thickness, and eosinophil infiltration were found to be decreased in all GMD groups compared with the negative control group. Conclusions : GMD is effective in atopic dermatitis by reducing imbalance of immune response of T cells (Th1 / Th2) and reducing skin tissue damage and inflammatory response.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.6
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• pp.1566-1571
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• 2008

Bone is a dynamic tissue that is constantly resorbed by osteoclasts and then replaced by osteoblasts. Osteoclasts, multinucleated cells of monocyte/macrophage lineage, are responsible for bone disorders, including osteoporosis and rheumatoid arthritis. In this study, we examined the effect of the curcumin on osteoclast survival and bone resorption. We found that curcumin significantly inhibited RANKL-mediated osteoclast survival. DAPI stainingrevealed that curcumin induced the apoptotic features of osteoclasts. Although curcumin did not suppress the phosphorylation of Akt and ERK in osteoclasts treated with RANKL, curcumin induced the cleavage of pro-caspase-9 and -3 its active forms. Also, curcumin inhibited the formation of actin rings of osteoclasts. RANKL-mediated bone resorption was inhibited by the addition of curcumin. Together with the results of this study, these findings suggest that the curcumin inhibited the survival of osteoclasts by activating caspase-9 and -3 and suppressed the bone resorptive activity. Thus, curcumin may be developed as antiresorptive drugs for the treatment of bone-related disorders.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.77-77
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• 2003

Coculture of HSC with bone marrow-derived mesenchymal stem cells (BM-MSCs) is one of used methods to increase cell numbers before transplant to the patients. However, because of difficulties to purify HSCs after coculture with BM-MSCs, it needs to develop a method to overcome the problem. In the present study, we have examined whether a culture insert placed over a feeder layer might support the expansion of HSCs within the insert. $CD34^+/ $ cells isolated from the umbilical cord blood by using midiMACS were divided into three groups. A group of 1 $\times$ $10^5$ cells were grown on a culture insert without feeder layer (Direct). The same number of HSCs was directly cocultured with BM-MSCs (Contact). The third group was placed onto an insert below which BM-MSCs were grown (Insert). To distinguish feeder cells from HSCs, BM-MSCs was pre-labeled fluorescently with PKH26 and 1 $\times$ $10^5$ cells were seeded in the culture dishes. After culture for 13 days, the expansion factor (x) of HSCs that were grown without feeder layer (Direct) was $26.6 \pm 8.4.$ In contrast, the number of HSCs directly cocultured with feeder layer was 59.6 $\pm$ 0.5 and that of HSCs cultured onto an insert was $46.9 \pm 8.4.$ The percentage of BM-MSCs cells remained being fluorescent was $97.9 \pm 0.3%$ after culture. Immune-phenotypically large proportion of cultured cells were founded to be differentiated into myeloid/monocyte progenitor cells. The ability of BM-MSCs, fetal lung, cartilage and brain tissue cells to support ex vivo expansion of HSCs was also examined using the insert. After 11 days of coculture with each of these cells, the expansion factor of HSCs was 15.0, 39.0, 32.0 and 24.0, respectively. Based upon these observations, it is concluded that the coculture method using insert is very effective to support ex vivo expansion of HSCs and to eliminate the contamination of other cells used to coculture wth HSCs.

• KSBB Journal
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• v.26 no.3
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• pp.248-254
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• 2011

Various biometerials have been researched and have been developed for treatment of some disease through transplantation to body. They have been evaluated by in vitro cytotoxicity test using some skin-derived cell lines for prediction of their biocompatibility in vivo. However, the results of experiments using mesenchymal or epithelial cells could not be considered in vivo immune reaction. In this study, we evaluated the biomaterial-elution (elute from high density polyethylene film) using some cell lines (L929, Jurkat, U937) in vitro, and then that results were compared with in vivo results from guinea pig sensitization test. In sensitization test, saline and elution of syringe could not induce erythema, but only DNCB (hypersensitive chemical) induce erythema at guinea pig sensitization test. In cell experiment, the cytotoxicity results of inflammatory cells (Jurkat; T lymphocyte, U937; monocyte) was no difference with L929 (fibroblast) in the overall trend. However, inflammatory cell lines were only secreted inflammatory cytokine (TNF-${\alpha}$, INF-${\gamma}$) in some materials (biomateriallution, FAC, DNCB). And the biomaterial-elution did not have toxicity to the cells, but it induced the inflammatory cytokines in inflammatory cell lines only. So, we were predicted inflammatory reaction through the cytokine resultes of inflammatory cell lines, and it was more correlated with in vivo results than cytotoxicity test. Therefore, we suggested that the inflammatory cytokine assay using inflammatory cell lines are more effective method in vitro for evaluation of biocompatibility of biomaterials or chemicals.