• Asian-Australasian Journal of Animal Sciences
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• v.24 no.3
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• pp.414-420
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• 2011

A study was conducted to investigate the effects of the combined stressor induced by high stocking density with feed restriction on immunological parameters such as leukocyte differential counts and cytokine expression in laying hens. A total of forty White Leghorn laying hens were randomly allotted into the control (12 kg of body weight/$m^2$) and the stress (44 kg of body weight/$m^2$) groups, and then birds of the stress group were given 75% of voluntary intake of the control birds for 12-d on a daily basis. There was a significant decrease in body weight without affecting the relative weights of the liver and spleen after 12-d of the combined stressor. In hematological values, no significant difference in leukocyte differential counts including heterophils (H), lymphocytes (L), monocytes and H:L ratio was observed between the two groups. In cytokines, hepatic lipopolysaccharide-induced tumor necrosis factor-${\alpha}$ (LITNF-${\alpha}$) and inducible nitric oxide synthase (iNOS) expression levels in the stress group were significantly (p<0.05) higher compared with those in the control group. However, the expression levels of interleukin (IL)-4 and IL-6 in the liver were not affected by the combined stressor. Splenic LITNF-${\alpha}$ expression in the combined stressor group was significantly (p<0.05) up-regulated compared with that in the control birds. However, the combined stressor did not affect splenic IL-4, IL-6 and iNOS expression. In conclusion, the combined stressor caused by high stocking density with feed restriction enhanced some pro-inflammatory cytokines including LITNF-${\alpha}$ and iNOS in lymphoid and non-lymphoid organs of birds, suggesting that altered cytokine expression to given stressors can be another parameter that can be used in assessing stress responses of birds.

• Journal of Acupuncture Research
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• v.32 no.3
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• pp.41-51
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• 2015

Objectives : This study was performed to investigate the effects of $LR_3$ and $SP_6$ acupuncture on renal damage in streptozotocin(STZ)-induced diabetic mice. Methods : ICR male mice were stabilized for a week and divided into four groups: a normal mice group(N), no-acupuncture diabetic mice group(Control), $LR_3$ acupuncture diabetic mice group($LR_3$), and $SP_6$ acupuncture diabetic mice group($SP_6$). Diabetes was experimentally induced by intraperitoneal injection of STZ(150 mg/kg) in citrate buffer(pH 4.5). For two weeks, $LR_3$ and $SP_6$ acupunctures were administered bilaterally at each point once a day. After two weeks, the animals' weight was measured and they underwent a laparotomy. Serum glucose and blood urea nitrogen(BUN) were measured from the blood taken from the heart. We measured glucose, reactive oxygen species(ROS), peroxynitrite($ONOO^-$) and thiobarbituric acid reactive substances(TBARS) in the kidney and compared expression levels of superoxide dismutases(SOD), glutathione peroxidase(GPx), nuclear factor-kappa B(NF-${\kappa}B$), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase(iNOS) and Interleukin-1 beta(IL-$1{\beta}$). Results : BUN significantly decreased in $LR_3$, $SP_6$ compared to the control group. $LR_3$ showed significantly decreased glucose compared to the control group. $LR_3$, $SP_6$ significantly decreased in ROS and $ONOO^-$ compared to the control group. $LR_3$ significantly decreased in TBARS compared to the control group. $SP_6$ significantly increased in expressions of SOD-1, catalase, and GPx compared to the control group. $LR_3$, $SP_6$ significantly decreased in COX-2 compared to the control group. $SP_6$ significantly decreased in IL-$1{\beta}$ compared to the control group. Conclusions : This study suggests that $LR_3$ acupuncture may be effective in controlling glucose and lipid peroxidation and that $SP_6$ acupuncture may have anti-oxidative and anti-inflammatory effects on renal damage in STZ-induced diabetic mice.

• Journal of Acupuncture Research
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• v.33 no.3
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• pp.29-43
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• 2016

Objectives : The purpose of this study was to investigate the effect of LR3 and SP6 acupuncture on liver damage of streptozotocin-induced diabetic mice. Methods : Male ICR mice were divided into four groups, consisting of the normal mice group(N), acupuncture-free diabetic mice group(Con), LR3-acupuncture diabetic mice group(LR3) and SP6-acupuncture diabetic mice group(SP6). The following measurements were taken: Body weight, food intake and water intake for 2 weeks; liver weight, and glucose levels in the serum and liver; ALT and AST in the serum; reactive oxygen species(ROS), reduced glutathione(GSH) and oxidized glutathione(GSSG) in the liver; and lastly, receptor for advanced glycation endproducts( RAGE), $N{\varepsilon}-carboxymethyl$ lysine(CML), $N{\varepsilon}-carboxyethyl$ lysine(CEL), phosphorylation of inhibitory kappa B alpha($p-I{\kappa}B{\alpha}$), nuclear factor-kappa B($NF-{\kappa}B$), activator protein-1(AP-1), cyclooxygenase-2(COX-2), inducible nitric oxide synthase(iNOS), tumor necrosis factor-alpha($TNF-{\alpha}$), ${\beta}-actin$, cytochrome c and caspase in the liver. Results : The liver weight and GSH/GSSG ratio were significantly increased in SP6 compared to Con. The glucose levels in the liver were significantly decreased in LR3 compared to Con. The generation of ROS and GSSG were significantly decreased in SP6 compared to Con. The expressions of RAGE, CML, AP-1, $TNF-{\alpha}$, cytochrome c and caspase 3 were significantly decreased in LR3 compared to Con. The expressions of $p-I{\kappa}B{\alpha}$, $NF-{\kappa}B$, AP-1, COX-2, iNOS and caspase 3 were significantly decreased in SP6 compared to Con. Conclusion : It is predicted that LR3 acupuncture is related to reduced glucose levels in the liver and expressions of AGE, and that, SP6 acupuncture is related to reduced oxidative stress-related transcription factors and inflammation-related proteins. Therefore, we suggest that LR3 and SP6 acupuncture have protective effects on the liver of streptozotocin-induced diabetic mice by preventing apoptosis.

• 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.

• Journal of Nutrition and Health
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• v.48 no.6
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• pp.459-467
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• 2015

Purpose: We developed a method to load lycopene into maltodextrin and cyclodextrin in an attempt to overcome the poor bioavailability and improve the anti-inflammatory effect of this polyphenol. Methods: Nanosized lycopenes were encapsulated into biodegradable amphiphillic cyclodextrin and maltodextrin molecules prepared using a high pressure homogenizer at 15,000~25,000 psi. Cell damage was induced by lipopolysaccharides (LPS) in a mouse macrophage cell line, RAW 264.7. The cells were subjected to various doses of free lycopene (FL) and nanoencapsulated lycopene (NEL). RT-PCR was used to quantify the tumor necrosis factor (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, inducible nitric oxide synthase (iNOS), and cyclooxigenase-2 (COX-2) mRNA levels, while ELISA was used to determine the protein levels of TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. Results: NEL significantly reduced the mRNA expression of IL-6 and IL-$1{\beta}$ at the highest dose, while not in cells treated with FL. In addition, NEL treatment caused a significant reduction in IL-6 and TNF-${\alpha}$ protein levels, compared to cells treated with a similar dose of FL. In addition, mRNA expression of iNOS and COX-2 enzyme in the activated macrophages was more efficiently suppressed by NEL than by FL. Conclusion: Overall, our results suggest that lycopene is a potential inflammation reducing agent and nanoencapsulation of lycopene can further improve its anti-inflammatory effect during tissue-damaging inflammatory conditions.

• Herbal Formula Science
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• v.17 no.2
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• pp.151-162
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• 2009

Herba Artemisiae Capillaris is the dried bud of Artemisia capillaris Thunb, which has been used for expelling heat to loosen the bowels and normalizing gallbladder function to cure jaundice in traditional oriental medicines. In the present study, we evaluated the anti-inflammatory effects of the aqueous extracts of Herba Artemisiae Capillaris (HAC) in LPS-activated Raw 264.7 cells. Cells were treated with $1\;{\mu}g/ml$ of LPS 1 h before adding HAC extract. Cell viability was determined by MTT assay, and the relative level of NO was measured with Griess reagent. TNF-$\alpha$, IL-$1{\beta}$, and IL-6 cytokines were detected by ELISA. During the entire experimental period, all three doses of HAC extract (0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity. LPS-activated cells showed increased NO levels and iNOS expressions compared to control. However, these increases were dramatically attenuated by treatment with HAC extract. Moreover, the inhibitory effects of HAC extract occurred in a dose-dependent manner. In addition, HAC extract reduced the translocation of $NF{\kappa}B$ into nuclear. HAC reduced production of IL-$1{\beta}$ and IL-6 by LPS, although it had no effects on TNF-$\alpha$. These results demonstrate that liquiritigenin exerts anti-inflammatory effects, which results from the inhibition of $NF{\kappa}B$ activation in macrophages, thereby decreasing production of iNOS and proinflammatory cytokines. Taken together, these results indicate that the aqueous extracts of Herba Artemisiae Capillaris warrant further development as an anti-inflammatory agent for the treatment of gram-negative bacterial infections.

• The Journal of Internal Korean Medicine
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• v.29 no.1
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• pp.130-148
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• 2008

Objective : This study was designed to investigate the effect of the water extract of Gamisamgibopae-tang(GMSGBPT), an oriental herbal formulation, on the growth of NCI-H460 and A549 human non-small-cell lung cancer cell lines. Methods : Cytotoxicity and cell morphology were evaluated by MTT assay and inverted microscope, respectively. Apoptosis was detected using agarose gel electrophoresis and flow cytometer. The expression levels of mRNAs and proteins of target genes were determined by RT-PCR and western blot analyses, respectively Result and Conclusion : We found that exposure of A549 cells to GMSGBPT resulted in the growth inhibition in a dose-dependent manner as measured by MTT assay, but GMSGBPTdid not affect the growth of NCI-H460 cells. The anti-proliferative effect of GMSGBPT treatment in A549 cells was associated with morphological changes, formation of apoptotic bodies and DNA fragmentation, and flow cytometry analysis confirmed that GMSGBPT treatment increased the populations of apoptotic-sub G1 phase. Growth inhibition and apoptotic cell death by GMSGBPT were connected with a up-regulation of cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) mRNA and protein in a tumor suppressor p53-independent fashion. However GMSGBPT treatment did not affect other growth regulation-related genes such as early growth response-1 (Egr-1), nonsteroidal anti-inflammatory drug (NSAID)-activated gene-1 (NAG-1), inducible nitric oxide synthase (iNOS), cyclooxygenases (COXs), telomere-regulatory factors in A549 orNCI-H460 cells. Taken together, these findings partially provide novel insights into the possible molecular mechanism of the anti-cancer activity of GMSGBPT.

• Korean Journal of Acupuncture
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• v.21 no.2
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• pp.47-67
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• 2004

Objectives : In the present study, the effect of Scolopendrid Water-Alcohol Extract (SWAE) applied to acupuncture point BL23 (Shinsu) on the neuropathic pain was examined. A common source of persistent pain in humans is the neuropathic pain. Anti-convulsant drugs are used to treat the neuropathic pain. In the oriental medicine, Scolopendrid was used for long time to treat convulsant syndrome and back pain, etc. Methods : On the bases of the Scolopendrid clinical application, the effect of SWAE applied to the acupuncture point was tested in the rat model of neuropathic pain. Neuropathic pain was induced by tight ligation of L5 spinal nerve. When rats developed pain behaviors, One hundred microliter of SWAE was applied into the ipsilateral BL23 point at a dose of 10 mg/ml under enflurane anesthesia. The foot withdraw latency of the hind limb was measured for an indicator of pain level after each manipulation. Results : SWAE injection increased the mechanical threshold of the foot in the rat model of neuropathic pain significantly for the duration of 4h, suggesting a partial alleviation of pain. SWAE applied to BL23 point produced a significant improvement of mechanical sensitivity of the foot lasting for at least 4h. However, neither contralateral BL23 point, ST25 (Chonchu) point, nor LR3 (Taechung) point produce as much increase of mechanical sensitivity as ipsilateral BL23 point. And, this increase of mechanical sensitivity was dose-dependent. The improvement of mechanical threshold was interpreted as an analgesic effect. In addition, the analgesic effect of Scolopendrid 4 mg/kg injection is equivalent to that of gabapentin 50 mg/kg injection. The relations between SWAE-induced analgesia and endogenous nitric oxide(NO), inducible NO synthase (iNOS)/neuronal NO synthase (nNOS) were also examined. Results were turned out that both NO production and nNOS/iNOS protein expression which are increased by nerve injury were suppressed by SWAE injection applied to BL23 point. Conclusions : The data suggest 1) that SWAE produces a potent analgesic effect on the neuropathic pain model in the rat and 2) that SWAE-induced analgesia modulate endogenous NO through the suppression of nNOS/iNOS protein expression.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.2
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• pp.310-318
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• 2010

Dioscorea bulbifera is one of the traditional medicinal herb. It commonly used in the treatment of hematemesis, epistaxis, tuberculous cervical lymphadenitis, laryngitis, acute infectious disease in East Asia. In the present study, we have demonstrated the anti-inflammatory effects of Dioscorea bulbifera MeOH extract (DBME) in macrophage cell line. To investigate mechanism of the anti-inflammatory activity, we examined the effects of the lipopolysaccaride (LPS)-induced production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), pro-inflammatory cytokines and expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), p-inhibitory ${\kappa}B{\alpha}$ (p-$I{\kappa}B{\alpha}$), and nuclear factor-${\kappa}B$ (NF-${\kappa}B$) in a murine macrophage cell line RAW 264.7. The RAW 264.7 cells were cultured in DMEM + serum medium for 24 hrs. After serum starvation for 24 hrs, the cells were treated with DBME 0.03, 0.10, 0.30 mg/$m{\ell}$ for 1 h, followed by stimulation with LPS (1 ${\mu}g/m{\ell}$) for activation of immune response. After treatment, cell viability was measured by MTT assay, and NO production was monitored by measuring the nitrite content in culture medium. The protein band of iNOS, COX-2, p-$I{\kappa}B{\alpha}$, and NF-${\kappa}B$ was determined by immunoblot analysis and levels of cytokine were analyzed by sandwich immunoassays. There were three experimental groups: carrageenan, DBME 0.3, 1.0 g/kg. Rats were administrated either carrageenan (40% PEG) or carrageenan + DBME (0.3, 1.0 g/kg body weight) for 4 days (p.o.). To induce acute paw edema, rats were injected 1% carrageenan (100 ${\mu}{\ell}$/rat, dissolved in sterilized saline). The effect of DBME in the carrageenan-induced rat paw edema. As results, DBME has an inhibitory effect on the production of NO, PGE2, TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 and on the expression of iNOS, COX-2, p-$I{\kappa}B{\alpha}$ and translocation of NF-${\kappa}B$ to nuclear from cytosol. In addition, DBME effectively inhibited the increases of paw edema induced by carrageenan treatment in vivo. These results suggest that DBME can inhibit production of pro-inflammatory mediators and might be a useful source for treatment of acute inflammatory disease.

• Journal of Pharmacopuncture
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• v.4 no.1
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• pp.123-124
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• 2001

Purpose. Free radicals are implicated in the pathophysiology of aging, ischemic injury and neurodegenerative disorders. To deform]no whether Hominis Placenta extract prevents $H_2O_2$-induced apoptosis, we have performed morphological and biochemical analyses for the detection of apoptotic phenomena in the pineal tumor cell line $PGT-{\beta}$ We have also peformed cytochemical and immunocytochemical analyses for the detection of changes in nitric oxide synthase (NOS) activity and estimated the expression . of apoptotic genes using reverse transcription-polymerase chain reaction (RT-PCR) Methods. $PGT-{\beta}\;cells$ were pretreated with Hominis Placenta extracts $(0,\;10^{-2}\;{\mu}g/ml)$ for 2 hours and then exposed to $H_2O_2\;(0,\;50\;{\mu}M)$ for 3 hours. Appearance of apoptotic characteristics were monitored using 4, 6-diamidino-2-phenylindole dihydrochloride (DAPI) staining assay, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay and flow cytometric analysis. NOS activity was measured by NADPH-diaphorase cytochemistry. Expression of inducible NOS (iNOS) and nuclear factor kappa B (NF k B) was assessed via immunocytochemistry. The expression of apoptotic genes was examined by RT-PCR. Results. After 3 flours of exposure to $H_2O_2$, it was shown that $PGT-{\beta}\;cells$ treated with $H_2O_2(50\;{\mu}M)$ exhibit classical apoptotic features and increases in NOS activity and caspase-3 expression. Treatment with Hominis Placenta extract resulted in a reduced occurrence of apoptotic features. DAPI staining, TUNEL and flow cytometric assays revealed decreases in the occurrence of nuclear fragmentation and in the sub-Gl fraction in the $PGT-{\beta}\;cells$ treated with Hominis Placenta extract. Cells treated with Hominis Placenta extract also showed lower activity of NADPH-diaphorase and immunoreactivities of both iNOS and NF k B than those of $H_2O_2$-treated cells which were not treated with Hominis Placenta extract. By RT-PCR, it was shown that the level of caspase-3 mRNA was derreased In the cells treated with Hominis Placenta . extract. Conclusions. This study shows that Hominis Placenta extract prevents $H_2O_2$-induced apoptosis in $PGT-{\beta}\;cells$; inhibitions of iNOS and caspnse-3 are possible mechanisms of the protection against apoptosis.