• Herbal Formula Science
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• v.19 no.1
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• pp.207-217
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• 2011

Objectives : In a previous study, we have shown that Gagam-Gongjin-Dan(GGD) has an inhibitory effect on the ovalbumin-induced immune responses and a hepatoprotective effect on actaminophen-induced liver injury in Balb/c Mice. However, the possible anti-inflammatory effect of GGD extract for inflammatory mediators was not reported. Therefore, the purpose of this study was to investigate an inhibitory effects of GGD extract against lipopolysaccharides(LPS) induced inflammatory mediators in mouse peritoneal macrophages. Methods : GGD extract was prepared by extracting with methanol for 7 days. The extract was freeze-dried following filtration through vacuum distillation system. Accumulated nitrite, an oxidative product of nitric oxide(NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin $E_2(PGE_2)$, interleukin-$1{\beta}$(IL-$1{\beta}$), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) were measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(COX-2) were measured by Western blot analysis. Results : GGD extract (50-$400\;{\mu}g$/ml) per se had no cytotoxic effect in LPS-stimulated peritoneal macrophages. GGD extract dose-dependently reduced NO, $PGE_2$, IL-$1{\beta}$ and TNF-${\alpha}$ production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with GGD extract in a dose dependent manner. Conclusions : These results suggest that GGD extract has an anti-inflammatory effect against LPS-induced inflammatory mediators in peritoneal macrophages, these properties may contribute to inflammation disease care.

• Herbal Formula Science
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• v.19 no.1
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• pp.195-205
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• 2011

Objectives : Ecklonia cava is brown alga(Laminariaceae) which grows is sea, it has antioxidant, diarrhea and anticoagulant effect. In this study, the effect of ethanol extract of Ecklonia cava (EC) on peptidoglycan(PGN) -induced NO production in RAW 264.7 cells was investigated. Methods : In the present study, IL-6 production was measured using the enzyme-linked immunosorbent assay(ELISA), prostaglandin $\E_2$($\PGE_2$) production was measured using the EIA kit, and inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) activation, as determined by western blot analysis and reverse transcription -polymerase chain reaction(RT-PCR). Results : EC inhibited PGN-induced NO and IL-6 production. Consistent with these observations, the protein expression of iNOS and COX-2 were inhibited by EC. Moreover, EC suppressed the phosphorylation of extracellular signal-regulated kinase(ERK) 1/2 in PGN-induced RAW 264.7. Conclusions : These results suggest that EC has inhibitory effects on PGN-induced $\PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

• Journal of Korean Medicine Rehabilitation
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• v.21 no.2
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• pp.1-13
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• 2011

Objectives : We investigated the effect of Haein-tang(Hairen-tang) on short-term memory and apoptosis in dentate gyrus of the gerbils with transient global ischemia. Methods : For the induction of cerebral ischemia model in mice, common carotid arteries of gerbils were occluded with aneurysm clips for 5 min. One day after operation, Haein-tang(Hairen-tang) was administrated orally injected once a day for 15 consecutive days. Gerbils were randomly divided into four group(n=10 in each group): sham-operation group, ischemia-induction group, ischemia-induction and 50 mg/kg Haein-tang(Hairen-tang)-treated group, ischemia-induction and 100 mg/kg Haein-tang(Hairen-tang)-treated group, and ischemia-induction and 200 mg/kg Haein-tang(Hairen-tang)-treated group. The effect of Haein-tang(Hairen-tang) on memory function was investigated by using step-down avoidance task. Apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) staining and immunohistochemistry for caspase-3. Western blot analysis for the expressions of Bax and Bcl-2 protein was also conducted. Results : 1. Haein-tang extract significantly enhanced short-term memory in step-down avoidance task and 100 mg/kg, 200 mg/kg Haein-tang-treated group. 2. Haein-tang extract significantly suppressed TUNEL-positive cells after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg Haein-tang-treated group. 3. Haein-tang extract significantly increased caspase-3 positive cells in the hippocampal dentate gyrus after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg Haein-tang-treated group. 4. Haein-tang extract significantly decreased Bax protein expressions in the hippocampus after transient global ischemia and 100 mg/kg, 200 mg/kg, Haein-tang-treated group. Haein-tang extract significantly increased Bcl-2 protein expressions in the hippocampal dentate gyrus after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg, Haein-tang-treated group. Haein-tang extract significantly decreased Ratio of Bax protein to Bcl-2 protein in the hippocampus after transient global ischemia and 100 mg/kg, 200 mg/kg Haein-tang-treated group. Conclusions : While Haein-tang(Hairen-tang) treatment improved short-term memory by suppressing on ischemia-induction apoptosis. In the present study, Haein-tang(Hairen-tang) shows protective effect on transient global ischemia.

• Journal of Periodontal and Implant Science
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• v.45 no.3
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• pp.101-110
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• 2015

Purpose: Sclerostin, an inhibitor of Wnt/${\beta}$-catenin signaling, exerts negative effects on bone formation and contributes to periodontitis-induced alveolar bone loss. Recent studies have demonstrated that serum sclerostin levels are increased in diabetic patients and that sclerostin expression in alveolar bone is enhanced in a diabetic periodontitis model. However, the molecular mechanism of how sclerostin expression is enhanced in diabetic patients remains elusive. Therefore, in this study, the effect of hyperglycemia on the expression of sclerostin in osteoblast lineage cells was examined. Methods: C2C12 and MLO-Y4 cells were used in this study. In order to examine the effect of hyperglycemia, the glucose concentration in the culture medium was adjusted to a range of levels between 40 and 100 mM. Gene expression levels were examined by quantitative reverse transcription-polymerase chain reaction and Western blot assays. Top-Flash reporter was used to examine the transcriptional activity of the ${\beta}$-catenin/lymphoid enhanced factor/T-cell factor complex. Tumor necrosis factor-alpha ($TNF{\alpha}$) protein levels were examined with the enzyme-linked immunosorbent assay. The effect of reactive oxygen species on sclerostin expression was examined by treating cells with 1 mM $H_2O_2$ or 20 mM N-acetylcysteine. Results: The high glucose treatment increased the mRNA and protein levels of sclerostin. High glucose suppressed Wnt3a-induced Top-Flash reporter activity and the expression levels of osteoblast marker genes. High glucose increased reactive oxygen species production and $TNF{\alpha}$ expression levels. Treatment of cells with $H_2O_2$ also enhanced the expression levels of $TNF{\alpha}$ and sclerostin. In addition, N-acetylcysteine treatment or knockdown of $TNF{\alpha}$ attenuated high glucose-induced sclerostin expression. Conclusions: These results suggest that hyperglycemia increases sclerostin expression via the enhanced production of reactive oxygen species and $TNF{\alpha}$.

• Journal of Periodontal and Implant Science
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• v.41 no.3
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• pp.149-156
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• 2011

Purpose: Globular adiponectin (gAd) is a type of adipocytokine, which is mainly produced by adipose tissue. It has been reported that gAd acts as a pro- as well as an anti-inflammatory factor. Interleukin (IL)-6 and IL-8 are pro-inflammatory cytokines. To investigate the role of gAd on periodontal tissues, the expression of adiponectin receptor 1 (AdipoR1) and the effect of gAd on the expression of IL-6 and IL-8 were investigated in periodontal ligament (PDL) and gingival fibroblasts. Methods: PDL and gingival fibroblasts were cultured from human periodontal tissues. gAd derived from Escherichia coli and murine myeloma cells were used. The expression of AdipoR1 was estimated by reverse transcription-polymerase chain reaction and western blot The expression of cytokines was measured by enzyme-linked immunosorbent assay. Results: PDL and gingival fibroblasts expressed both mRNA and protein of AdipoR1. gAd derived from E. coli increased the production of IL-6 and IL-8, but polymyxin B, an inhibitor of lipopolysaccharide (LPS), inhibited IL-6 and IL-8 production induced by gAd in both types of cells. gAd derived from murine myeloma cells did not induce IL-6 and IL-8 production in those cells. gAd derived from E. coli contained higher levels of LPS than gAd derived from murine myeloma cells. LPS increased production of IL-6 and IL-8 in PDL and gingival fibroblasts, but pretreatment of cells with gAd derived from murine myeloma cells did not inhibit LPS-induced IL-6 and IL-8 expression. Conclusions: Our results suggest that PDL and gingival fibroblasts express AdipoR1 and that gAd does not act as a modulator of IL-6 and IL-8 expression in PDL and gingival fibroblasts.

• The Korea Journal of Herbology
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• v.31 no.5
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• pp.71-78
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• 2016

Objectives : Diabetic nephropathy is one of the most common chronic complications of diabetes and a leading cause of end-stage renal failure in the world. Mesangial cell proliferation is known as the major pathologic features such as glomerulosclerosis and renal fibrosis. Wiryeongtang (WRT) is a well-known traditional herbal formula as therapeutic agents for chronic edema and dysuresia of renal homeostasis. In the present study, we investigated whether WRT inhibits high glucose (HG)-induced renal dysfunction by TGF-β/Smads signal regulation in cultured mesangial cells.Methods : Inhibitory effect of WRT (10-50 ㎍/ml) on HG-stimulated mesangial cells proliferation and dysfunction were evaluated by [³H]-thymidine incorporation, Western blot, and RT-qPCR.Results : WRT significantly decreased HG-accelerated thymidine incorporation in human renal mesangial cell in a dose-dependent levels. WRT induced down-regulation of cyclins/CDKs and up-regulation of CDK inhibitor, p21waf1/cip1 and p27kip1 expression. In addition, HG enhanced expression of dysfunction biomarker such as collagen IV and CTGF, which was markedly attenuated by WRT. WRT decreased TGF-β1 and Smad-2/Smad-4 expression, whereas increased Smad-7 expression under HG. Furthermore, WRT inhibited HG-induced inflammatory factors level such as ICAM-1 and MCP-1 as well as NF-κB p65 nuclear translocation and intracellular ROS production.Conclusions : These results suggested that WRT may alleviate mesangial proliferation and inflammation possibly involved in renal fibrotic process, further diabetic nephropathy through disturbing TGF-β1/Smad signaling and NF-κB/ROS pathway. Thus, WRT might prove to be effective in the treatment of renal dysfunction leading to diabetic nephropathy.

• The Korea Journal of Herbology
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• v.27 no.5
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• pp.99-107
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• 2012

Objectives : This study was conducted to investigate the anti-obesity effects of mixed water extract of Plantaginis Semen & Poria (CJB) on obese rats induced with high fat diet. Method: Male Sprague-Dawley rats were divided into three groups; Normal group, high-fat (HF) group, HF+CJB(100 mg/kg, P.O.) for 8 weeks. The body weight, food intake and weights of adipose tissues were measured, respectively. Lipid profiles in serum were analyzed by automatic analyzer of blood. Obese marker proteins and the changes of NPY and LR immunoreactivities in hypothalamus were analyzed by Western blot and immunohistochemistry. Results : CJB significantly reduced body weight, food intake, adipose tissue weights compared to HF group. Serum triglyceride and total cholesterol were significantly higher in HF group than in Normal group however, CJB significantly lowered those of HF group. HDL-cholesterol level in CJB groups was elevated compared to HF group. The pAMPK in hypothalamus were decreased in that of HF group and that of CJB group decreased. Inversely, ACC was increased in HF group and that of CJB groups decrease. Expression of $PPAR{\gamma}$ in hypothalamus was increased by CJB treatment. However, $PPAR{\alpha}$ levels in CJB group were decreased compared to HF group. The expressions of NPY and LR in PVN and ARC of hypothalamus were decreased in CJB group, respectively. Conclusion : Administration of CJB can play anti-obesity through regulations of NPY and LR activities and obesity marker proteins in obese rat's hypothalamus.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.39-47
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• 2011

Objective: The roots, leaves, flowers, stems and seeds of Cirsium japonicum var. ussuriense are often used in treatment of human diseases such as hemorrhage, blood congestion and inflammation. Focusing our attention on natural and bioavailable sources of antioxidants and anti-inflammation, we undertook to investigate the antioxidant and anti-inflammatory properties of Cirsium japonicum var. ussuriense used as a folk medicine in Korea. Methods: The extracts of the leaves, stems, flowers, seeds and roots from C. japonicum var. ussuriense were prepared by extracting with water or 80% ethanol. Total flavonoids and polyphenols were measured by a colorimetric assay. The free radical scavenging activity of the extract was analyzed by the DPPH (1,1-diphenyl-2-picryl hydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and Griess reagent assay. An oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The level of prostaglandin $E_2$ ($PGE_2$) was measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were measured by Western blot analysis. Results: Total flavonoid and polyphenol amounts of the leaves (CLE) and flowers (CFE) showed higher than those of the seed extract (CSE), stem extract (CSTE) and roots (CRE). CLE and CFE also showed the high antioxidant activities such as DPPH, NO-like and ABTS radical scavenging activity. An antioxidant activities of these water extracts showed higher than those of 80% ethanol extracts. We investigated the anti-inflammatory effects of CLE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. CLE significantly suppressed the levels of the inflammatory mediators such as NO and prostaglandin $E_2$ ($PGE_2$) in dose dependant. Furthermore, the levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with CLE extract in a dose dependent manner. Conclusions: These results suggest that CLE water extract has a higher anoxidant and anti-inflammatory activity, these properties may contribute to the oxidative and inflammatory related disease care.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.32 no.4 s.59
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• pp.241-247
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• 2006

This work was carried out to investigate the anti-inflammatory effects of Paulownia coreana Uyeki on abirritant, atopy and acne skin. Paulownia coreana Uyeki has been used as a traditional medicine having anti-febrile, anti-inflammation effect in Korea, Paulownia coreana Uyeki loaves were extracted with 70% EtOH. Its superoxide anion radical scavenging activity and inhibitory effect on LPS-induced NO production were examined. The extract inhibitied the generation of NO and $PGE_2$ induced by LPS in the macrophage cell line (Raw 264.7). Consistent with the inhibitory effects on No and $PGE_2$ generation, the extract inhibited expression of iNOS and COX-2. In further study, it was found that the extract prevented $IkB-{\alpha}$ degradation, as demonstrated by western blot analysis of $IkB-{\alpha}$ protein level. However, the extract treatment did not affect cell viability at $100{\mu}g/mL$ concentration in both human skin fibroblast and Raw 264.7 cells in vitro. Thus, the present study suggests that Paulownia coreana Uyeki leaves extract have significant anti-inflammatory activity and potential as an anti-irritation material.

• Food Science and Preservation
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• v.21 no.1
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• pp.107-113
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• 2014

The present study was designed to investigate the antihyperlipidemic effect of Coconopsis lanceolata extract in C57BL/6J mice. The mice were divided into four groups: normal diet group (ND), high fat diet group (HFD), positive control group with 0.05% metformin (PC), Coconopsis lanceolata extract group (UCL). After 5 weeks of feeding, average body weight of the UCL group mice was slightly decreased, while that of the HFD group significantly increased) Also, liver and adipose tissue weights in the UCL group significantly increased. The levels of trigliceride (TG) and total in the plasma of UCL-supplemented group were significantly lower than those of high fat diet group. On the other hand, HDL-cholesterol level was increased. Expression level of proteins related with adipogenesis such as SREBP-1c, ACC, and FAS in the liver of the UCL group mice was much lower comparing with the HFD group mice. In conclusion, the results showed that the Coconopsis lanceolata extract possesses significant antihyperlipidemic effects in C57BL/6J mice.