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Effect of Sinapis alba L. on expression of interferon-gamma and interleukin-4 production in anti-CD3/anti-CD28-stimulated CD4(+) T cells (CD4+ T cells에서 백개자가 IFN-$\gamma$와 IL-4 생성에 미치는 영향)

  • Park, Dae-Jung;Lee, Jang-Cheon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.25 no.2
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    • pp.129-136
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    • 2010
  • Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal macrophage immune responses have not yet been determined. In this study, we examined the effect of SA on the production of Th1/Th2 cytokines. Methods : Splenocytes were isolated from naive C57BL/6 mice. Cells were enriched for CD4+ cell populations by first staining the cells with anti-CD4 (BD PharMingen, Calif, USA). CD4+ T cells were selected on a (CS) column, and the flow-through was collected as CD4+ T cells. Isolated cells were activated by overnight incubation on 24-well plates coated with $1{\mu}g/mL$ anti-CD3, $1{\mu}g/mL$ anti-CD28 and with SA ($100{\mu}g/mL$). Primary macrophages were collected from the peritoneal cavities of mice (8-week-old female C57BL/6). The peritoneal macrophages were washed and plated with RPMI-1640 overnight for the experiments. After 48-hours cultures, samples were centrifuged at 2000 rpm for 10 minutes, and the supernatants were stored at $-80^{\circ}C$. Mouse IL-4, IFN-$\gamma$ and TNF-$\alpha$ were quantified using ELISA kits (BioSource International, Camarillo, Calif, USA) according to the manufacturer's protocols. Results : SA at 100ug/ml decreased the generation of Th1 cytokine (IFN-$\gamma$) by 0.5-fold. However, SA has no effect on Th2 (IL-4) production. Conclusions : These results suggest that SA may play an important role in the control of T-cell-mediated autoimmunity by down-regulation of Th1 cytokine (especially IFN-$\gamma$, TNF-$\alpha$). These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.

Effect of Sodium Butyrate on Blood Glucose, Serum Lipid Profile and Inflammation in Streptozotocin-induced Diabetic Mice (스트렙토조토신으로 유도한 당뇨마우스에서 Sodium Butyrate의 혈당, 혈청 지질 성상 및 염증 억제에 미치는 영향)

  • Yun, Jung-Mi
    • The Korean Journal of Food And Nutrition
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    • v.28 no.2
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    • pp.171-177
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    • 2015
  • Sodium butyrate is a short-chain fatty acid derivative found in foods, such as Parmesan cheese and butter and is produced by anaerobic bacteria fermentation of dietary fibers in the large intestine. There have been reports that butyrate prevented obesity, protected insulin sensitivity, and ameliorated dyslipidemia in dietary obese mice. This study investigated the effects of sodium butyrate on fasting blood glucose level and serum lipid profile in streptozotocin(STZ)-induced diabetic mice. Male C57BL/6 mice were fed AIN-93G for four weeks prior to intraperitoneal injections with STZ (100 mg/kg body weight). Diabetic mice had supplements of 5% sodium butyrate for four weeks. The 5% sodium butyrate diet significantly improved fasting blood glucose level and lipid profile in STZ-induced diabetic mice. Inflammation has been recognized to decrease beta cell insulin secretion and increase insulin resistance. Circulating cytokines can directly affect beta cell function, leading to secretory dysfunction and increased apoptosis. Thus, anti-inflammatory therapies represented a potential approach for the therapy of diabetes and its complications. In this animal study, the 5% sodium butyrate supplementation also inhibited inflammatory cytokine production in STZ-induced diabetic mice. These results suggested that sodium butyrate can be a potential candidate for the prevention of diabetes and its complications.

Aucklandiae Radix Ameliorates Scopolamine-induced Memory Impairment in Mice (Scopolamine 유발 기억력 손상 마우스 모델에서 목향(木香)의 기억력 개선 및 항산화 효과)

  • Park, Na-eun;Han, Da-young;Kim, Sang-ho;Chung, Dae-kyoo
    • Journal of Oriental Neuropsychiatry
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    • v.28 no.2
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    • pp.123-136
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    • 2017
  • Objectives: The objective of this study is to investigate the anti-amnesic effects of AR, Aucklandiae Radix, ground powder on scopolamine (Sco)-induced memory impairment in mice (C57BL/6) through its favorable acetylcholine (ACh) and acetylcholinesterase (AChE) activity, Choline acetyltransferase (ChAT) mRNA expression, and antioxidant effect. Methods: Six groups, a total of 20 intact or 100 Sco treated mice, were selected based on their body weights and were used in this study. Half of the mice in each group were used for the passive avoidance task test and the measurements of hippocampus ACh content, AChE activity and ChAT mRNA expression. The remaining half of the mice in each group were used for the Morris water maze test and cerebral antioxidant defense system measurement. Results: Marked decreases in step-through latency times in the passive avoidance task test and increases in escape latency times in the Morris water maze test were observed with decreases in the hippocampus ACh content and ChAT mRNA expression, and increases in the hippocampal AChE activities, as a result of Sco intraperitoneal treatment, in the present study. In addition, destruction of the cerebral cortex antioxidant defense systems was observed in Sco control mice as compared with intact vehicle control mice. However, 28 days of continuous oral pre-treatment with AR ground powder at doses of 400, 200 and 100 mg/kg markedly and dose-dependently inhibited the Sco treatment-related amnesia. Conclusions: The results prove that oral administration of AR ground powder reduces Sco-induced memory impairment. This is because it can preserve ACh, related to ChAT mRNA expression, cause AChE inhibition, and activate the cerebral antioxidant defense system.

Anti-parkinsonian effect of Cyperi Rhizoma via inhibition of neuroinflammatory action (향부자(香附子)의 염증 억제 작용을 통한 항파킨슨 효과)

  • Kim, Hyo Geun;Sim, Yeomoon;Oh, Myung Sook
    • The Korea Journal of Herbology
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    • v.28 no.5
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    • pp.21-28
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    • 2013
  • Objectives : The aim of this study was to investigate the neuroprotective effects and mechanisms of Cyperi Rhizoma extracts (CRE) using in vitro and in vivo models of Parkinson's disease (PD). Methods : We evaluated the neuroprotective effect of CRE against 1-methyl-4-phenylpyridinium (MPP+) toxicity using tyrosine hydroxylase immunohistochemistry (IHC) in primary rat mesencephalic dopaminergic neurons. In addition, the effect of CRE was evaluated in mice PD model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). For evaluations, C57bl/6 mice were orally treated with CRE 50 mg/kg for 5 days and were injected intraperitoneally with MPTP (20 mg/kg) at 2 h intervals on the last day. To identify the CRE affects on MPTP-induced neuronal loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) and striatum of mice, the behavioral tests and IHC analysis were carried out. Also, we conducted nitric oxide (NO) and tumor necrosis factor-alpha (TNF-${\alpha}$) assay in dopaminergic neurons and IHC using glial markers in SNpc of mice to assess the anti-inflammation effects. Results : In primary mesencephalic culture system, CRE protected dopaminergic cells against $10{\mu}M$ MPP+-induced toxicity at 0.2 and $1.0{\mu}g/mL$. In the behavior tests, CRE treated group showed improved motor deteriorations than those in the MPTP only treated group. CRE significantly protected striatal dopaminergic damage from MPTP-induced neurotoxicity in mice. Moreover, CRE inhibited productions of NO and TNF-${\alpha}$ in dopaminergic culture system and activation of astrocyte and microglia in SNpc of the mice. Conclusion : We concluded that CRE shows anti-parkinsonian effect by protecting dopaminergic neurons against MPP+/MPTP toxicities through anti-inflammatory actions.

Anti-cancer Effects of Cultivated Orostachys japonicus on Human Colon Cancer Cell Line SW480 (인체대장암세포주 SW480에 대한 재배 와송의 항암효과 연구)

  • Park, Sookyoung;Won, Jinyoung;Park, Kanghui;Hong, Yonggeun
    • Journal of Life Science
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    • v.28 no.7
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    • pp.819-826
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    • 2018
  • Orostachys japonicus (OJ) is a medicinal herb with immunoregulatory, anti-aging, anti-oxidative, and many other therapeutic properties. The purpose of this study was to elucidate the anti-cancer property of cultivated OJ. SW480 cell viability was significantly reduced by cumulative exposure to OJ extract. We also observed inhibitory effects of OJ after 72 hr through the growth and migration of SW480 cells using scratch assay. SW480 cells in OJ-free medium began to move into the scratch site at 24 hr; however, cells in medium containing OJ did not migrate into the scratch site until 48 hr. Male C57BL/6 mice (4 weeks old) were orally administered OJ extract for 31 days before injection of SW480 cells. At 7, 14, and 28 days after subcutaneous injection of SW480 cells, tumor weight and volume were analyzed. The body weight of the OJ-treated group was continuously increased during administration of the OJ extract relative to the control group. Injection of SW480 cells caused a reduction in body weight in all groups; however, the OJ-treated group exhibited a significant increase in body weight after 14 days. Tumor weight and volume were lower in the OJ-treated group than in the control group after 28 days. Although these results suggest that OJ suppresses the proliferation and migration of human colon cancer cells, additional studies are required to provide preclinical evidence before launching clinical trials evaluating OJ as an anti-cancer biohealth product.

In Vivo Effects of CETP Inhibitory Peptides in Hypercholesterolemic Rabbit and Cholesteryl Ester Transfer Protein-Transgenic Mice

  • Cho, Kyung-Hyun;Shin, Yong-Won;Choi, Myung-Sook;Bok, Song-Hae;Jang, Sang-Hee;Park, Yong-Bok
    • BMB Reports
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    • v.35 no.2
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    • pp.172-177
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    • 2002
  • We previously reported that cholesteryl ester transfer protein (CETP) inhibitory peptides (designated $P_{28}$ and $P_{10})$ have anti-atherogenic effects in hypercholesterolemic rabbits (Biochim. Biophys. Acta (1998) 1391, 133-144). To further investigate those effects, we studied rabbit plasma that was collected after 30 h of a $P_{28}$ or $P_{10}$ injection. We found that there is a strong correlation between the in vivo CETP inhibition effects and alterations of lipoprotein particle size distribution in rabbit plasma, as determined on an agarose gel electrophoresis and gel filtration column chromatography. In vivo effects of the peptide were observed again in C57BL/6 mice that expressed simian CETP. The $P_{28}$ or $P_{10}$ peptide ($7\;{\mu}g/g$ of body weight) that was dissolved in saline was injected subcutaneously into the mice. The $P_{28}$ injection caused the partial inhibition of plasma CETP activity up to 50%, decreasing the total plasma cholesterol concentration by 30%, and increasing the ratio of HD/total-cholesterol concentration by 150% in the CETP-transgenic (tg) mice. The CETP inhibition by the $P_{28}$ or $P_{10}$ made alterations that modulated the size re-distribution of the lipoproteins in the blood stream. Particle size of the very low (VLDL) and low density lipoproteins (LDL) from the peptide-injected group was highly decreased compared to the saline-injected group (determined on the gel filtration column chromatography). In contrast, The HDL particle size of the $P_{28}$-injected group increased compared to the control group (saline-injected). The expression level of the CETP mRNA of the $P_{28}$-injected CETP-tg mouse appeared lower than the saline-injected CETP-tg mouse. These results suggest that the injection of the CETP inhibitory peptide could affect the CETP expression level in the liver by influencing lipoprotein metabolism.

Anti-Obesity Effects of Salted and Unsalted Doenjang Supplementation in C57BL/6J Mice Fed with High Fat Diet (식이유도 비만 Mice에서 된장 및 무염된장의 항비만 효과)

  • Bae, Cho-Rong;Kwon, Dae Young;Cha, Youn-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1036-1042
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    • 2013
  • The aim of the present study is to elucidate the anti-obesity effects of Doenjang with and without salt in C57BL/6J mice. For the analysis, a total of forty mice were randomly divided into four groups: normal diet group (ND), high-fat diet group (HD), high-fat diet supplemented with 20% Doenjang group (DJ), high-fat diet supplemented with 20% unsalted Doenjang group (NS). During the study period, food intake and body weight were measured daily and weekly, respectively. The animals were sacrificed after 12 weeks. Body weight gain, epididymal fat pad weight and serum triglyceride levels of DJ group were found to be significantly lower than those of the HD and NS groups. Serum total-cholesterol levels of DJ and NS groups were significantly lower as compared to the HD group. There were significant decreases in plasma insulin and leptin levels in DJ group compared with the HD and NS groups. We did not observe any significant changes in the expression of hepatic lipogenic-related gene $PPAR{\gamma}$ among the HD, DJ and NS groups. However, ACC expression was found to be significantly decreased in DJ group. Lipolysis-related gene ($PPAR{\alpha}$ and CPT-1) expression was significantly higher in the DJ group as compared to HD and NS groups. In conclusion, the results of this study showed that Doenjang supplementation lowers body weight gain and improves obesity-related parameters.

Purification and Characterization of a Recombinant Pea Chloroplastic Fructose-1, 6-bisphosphatase

  • Shin, Eun-Hye;Yoo, Yong-Cheol;Lee, Sang-Won;Hahn, Tae-Ryong
    • Journal of Applied Biological Chemistry
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    • v.44 no.4
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    • pp.167-172
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    • 2001
  • A cDNA fragment encoding the chloroplastic fructose-1, 6-bisphosphatase (FBPase) was cloned via PCR from the cDNA library of pea leaves. The cloned cDNA, about 1.05 kbp without signal sequence, was introduced into a pET-28a vector for expression in E. coli strain BL21(DE3)pLysS. The recombinant FBPase was purified through $Ni^+-NTA$ affinity chromatography and characterized. Molecular mass of the monomer was about 42,000. Enzymatic activity of the purified enzyme as the native pea chloroplastic FBPase was the highest at alkaline pH (pH 9.0). The recombinant enzyme was activated by a reducing agent DTT and was insensitive to AMP. The activation energy (Ea) and Arrehenius frequency factor were 42.67 kcal/mol and $2.65{\times}10^{14}/s$, respectively, slightly higher than those of the native enzyme. $K_M$ and $V_{max}$ were $99.98{\mu}M$ and $52.9{\mu}M/min$, respectively, which were comparable with the native enzyme.

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Anti-Depressive Effects of OnDam-Tang with Addition of Linderae Radix (ODT-L) after Chronic Immobilization Stress in C57BL/6 Mice (우울증 유발 생쥐에서 온담탕가오약(溫膽湯加烏藥)의 항우울 효과)

  • Lee, Eun Hee;Jung, In Chul
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.4
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    • pp.403-410
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    • 2014
  • The purpose of this study was to examine the anti-depressive effects of OnDam-Tang with addition of Linderae Radix (ODT-L) on the animal model of depression induced by chronic immobilization stress. Depression model was made by chronic immobilization stress for 2 hours for 21 days. And we performed forced swimming test, analysis of the neurotransmitter and immunohistochemical staining, measured expression levels of serotonin in the brain. ODT-L has decreased immobilization time in forced swimming test. ODT-L has increased amount of melatonin in the brain. ODT-L has increased expression levels of serotonin in the brain. ODT-L prevented damage in the hippocampal region. ODT-L has reduced the expression level of CRF receptors in in hippocampus region. These results suggest that ODT-L may have anti-depressive effects on depression.

Effect of Ethanolic Extract of Silkworm on Reactive Oxygen Species Formation in vivo (누에의 에탄올 추출물이 체내 활성산소 생성에 미치는 영향)

  • Kang, Young Kook;Choi, Min Joo;Nam, Sang Ho
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.4
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    • pp.379-383
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    • 2014
  • The purpose of this study is to investigate the effects of silkworm extracts (SWE) on reactive oxygen species formation in mice (C57BL/6). Mice were administrated intraperitoneally with SWE (20 mg/kg/day) for 14 days. All animals were sacrificed 24 hours after the last SWE treatment and then extracted the blood and brain tissue in mouse. The researcher measured several parameters related to reactive oxygen species formation, malondialdehyde (MDA) and hydrogen peroxide ($H_2O_2$) contents in serum, whole brain, cerebral cortex and cerebellum. The results showed that MDA content of pre-SWE treatment was decreased significantly in serum, mitochondrial and cytosolic fraction of whole brain and cerebellum (P<0.01). The $H_2O_2$ content of pre-SWE treatment was decreased significantly in mitochondrial fraction of whole brain, cerebral cortex and cerebellum (P<0.01). These results suggest that SWE plays an important role for inhibition of oxidative damage of cells as well as antioxidant effect, aging delay and cells protected from irradiation.