• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.83-83
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• 2003

In our previous study, glucose deprivation was reported to induce the potentiated death and ATP loss in immunostimulated astroglia. And this vulnerability to glucose deprivation was due to overproduction of nitric oxide (NO) and hydrogen peroxide (H$_2$O$_2$). In the present study, the role of extracellular signal-regulated kinase 1/2 (ERK1/2) in the glucose deprivation-induced death of immunostimulated astroglia was examined. We showed that immunostimulation with LPS+IFN-ν activated the ERKl/2 signal pathway and produced a large amount of NO and H$_2$O$_2$. Generation of NO and H$_2$O$_2$ in immunostimulated astroglia was mediated via ERK1/2 signal pathways, since addition of the ERK kinase (MEKl) inhibitor PD98059 reduced NO and H$_2$O$_2$production. ERK1/2 activation-mediated NO and H$_2$O$_2$ production is due to an activation of iNOS and NADPH oxidase, respectively. Finally, we found that glucose deprivation caused ATP depletion and the augmented death in immunostimulated astroglia, which was also prevented by PD98059 treatment. These results demonstrate that the ERK1/2 signal pathways play an important role in glucose deprivation induced the death in immunostimulated astroglia by regulating the generation of NO and H$_2$O$_2$.

• Biomolecules & Therapeutics
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• v.24 no.2
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• pp.184-190
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• 2016

Rosa rugosa Thunb, a deciduous shrub of the genus Rosa, has been widely used to treat stomach aches, diarrhoea, pain, and chronic inflammatory disease in eastern Asia. In recent years, our research team has extensively studied the Rosa rugosa flower extract, and specifically undertook pharmacological experiments which have optimized the extraction process. Our methods have yielded a standard extract enriched in phenolic compounds, named PRE. Herein, we expand our efforts and evaluated the anti-inflammatory activity of PRE on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophages. PRE significantly inhibited production of nitric oxide (NO), prostaglandin $E_2(PGE_2)$, tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and interleukin $1{\beta}$ (IL-$1{\beta}$), as well as expression of their synthesizing enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase2 (COX-2). Furthermore, PRE inhibited activity of mitogen-activated protein kinases (MAPK) as well as nuclear factor-kappa B (NF-${\kappa}B$) signaling pathway. Our findings are the first to explain the anti-inflammatory mechanism by PRE in LPS-stimulated macrophages. Given these results, we propose that PRE has therapeutic potential in the prevention of inflammatory disorders.

• Biomolecules & Therapeutics
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• v.22 no.4
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• pp.295-300
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• 2014

The actin cytoskeleton plays an important role in macrophage-mediated inflammatory responses by modulating the activation of Src and subsequently inducing nuclear factor (NF)-${\kappa}B$ translocation. In spite of its critical functions, few papers have examined how the actin cytoskeleton can be regulated by the activation of toll-like receptor (TLR). Therefore, in this study, we further characterized the biological value of the actin cytoskeleton in the functional activation of macrophages using an actin cytoskeleton disruptor, cytochalasin B (Cyto B), and explored the actin cytoskeleton's involvement in morphological changes, cellular attachment, and signaling events. Cyto B strongly suppressed the TLR4-mediated mRNA expression of inflammatory genes such as cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-${\alpha}$, and inducible nitric oxide (iNOS), without altering cell viability. This compound also strongly suppressed the morphological changes induced by lipopolysaccharide (LPS), a TLR4 ligand. Cyto B also remarkably suppressed NO production under non-adherent conditions but not in an adherent environment. Cyto B did not block the co-localization between surface glycoprotein myeloid differentiation protein-2 (MD2), a LPS signaling glycoprotein, and the actin cytoskeleton under LPS conditions. Interestingly, Cyto B and PP2, a Src inhibitor, enhanced the phagocytic uptake of fluorescein isothiocyanate (FITC)-dextran. Finally, it was found that Cyto B blocked the phosphorylation of vasodilator-stimulated phosphoprotein (VASP) at 1 min and the phosphorylation of heat shock protein 27 (HSP27) at 5 min. Therefore, our data suggest that the actin cytoskeleton may be one of the key components involved in the control of TLR4-mediated inflammatory responses in macrophages.

• Biomolecules & Therapeutics
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• v.24 no.6
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• pp.610-615
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• 2016

Quercetin, a flavonol, has been reported to exhibit a wide range of biological properties including anti-oxidant and anti-inflammatory activities. However, pharmacological properties of quercetin-3-O-${\beta}$-D-glucuronide (QG), a glycoside derivative of quercetin, have not been extensively examined. The objective of this study is to elucidate the anti-inflammatory property and underlying mechanism of QG in lipopolysaccharide (LPS)-challenged RAW264.7 macrophage cells in comparison with quercetin. QG significantly suppressed LPS-induced extracellular secretion of pro-inflammatory mediators such as nitric oxide (NO) and $PGE_2$, and pro-inflammatory protein expressions of iNOS and COX-2. To elucidate the underlying mechanism of the anti-inflammatory property of QG, involvement of MAPK signaling pathways was examined. QG significantly attenuated LPS-induced activation of JNK and ERK in concentration-dependent manners with a negligible effect on p38. In conclusion, the present study demonstrates QG exerts anti-inflammatory activity through the suppression of JNK and ERK signaling pathways in LPS-challenged RAW264.7 macrophage cells.

• Journal of Ginseng Research
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• v.40 no.2
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• pp.196-202
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• 2016

Background: Ginsenoside Rd (GSRd), a main component of the root of Panax ginseng, exhibits anti-inflammation functions and decreases infarct size in many injuries and ischemia diseases such as focal cerebral ischemia. M1 Macrophages are regarded as one of the key inflammatory cells having functions for disease progression. Methods: To investigate the effect of GSRd on renal ischemia/reperfusion injury (IRI) and macrophage functional status, and their regulatory role on mouse polarized macrophages in vitro, GSRd (10-100 mg/kg) and vehicle were applied to mice 30 min before renal IRI modeling. Renal functions were reflected by blood serum creatinine and blood urea nitrogen level and histopathological examination. M1 polarized macrophages infiltration was identified by flow cytometry analysis and immunofluorescence staining with $CD11b^+$, $iNOS^+$/interleukin-12/tumor necrosis factor-${\alpha}$ labeling. For the in vitro study, GSRd ($10-100{\mu}g/mL$) and vehicle were added in the culture medium of M1 macrophages to assess their regulatory function on polarization phenotype. Results: In vivo data showed a protective role of GSRd at 50 mg/kg on Day 3. Serum level of serum creatinine and blood urea nitrogen significantly dropped compared with other groups. Reduced renal tissue damage and M1 macrophage infiltration showed on hematoxylin-eosin staining and flow cytometry and immunofluorescence staining confirmed this improvement. With GSRd administration, in vitro cultured M1 macrophages secreted less inflammatory cytokines such as interleukin-12 and tumor necrosis factor-${\alpha}$. Furthermore, macrophage polarization-related pancake-like morphology gradually changed along with increasing concentration of GSRd in the medium. Conclusion: These findings demonstrate that GSRd possess a protective function against renal ischemia/reperfusion injury via downregulating M1 macrophage polarization.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.2 s.33
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• pp.77-88
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• 2007

Objectives : This experimental study was performed to investigate the effects of Yeouigeumhwang-san(YUGHS) on anti-inflammation and anti-Propionibacterium acnes. Methods : The cytotoxicity of YUGHS about viability of Raw 264.7 cell was tested by using a colorimetric tetrazolium assay(MTT assay). To investigate the anti-inflammatory effets of YUGHS on LPS-induced macrophage Raw 264.7 cell, we used ELISA kit and Western blots. Inhibitory effects of YUGHS on Propionibactrium acnes were investigated by using paper disk diffusion method. Results : 1. YUGHS has no cytotoxicity under 50 ${\mu}g/ml$ concentration but over 50 ${\mu}g/ml$ has a little cytotoxicity in Raw 264.7 cell. 2. Concentration of 100 ${\mu}g/ml$ YUGHS inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of YUGHS did not inhibit the production of $TNF-{\alpha}$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of YUGHS significantly inhibited the production of $PGE_2$ in the Raw 264.7 cell stimulated with LPS. 5. YUGHS did not inhibit the expression of COX-2 but concentration of 50 ${\mu}g/ml$ YUGHS inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. YUGHS has the effect of blocking $NF-{\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell 7. YUGHS did not have the inhibitory effect of Propionibactrium acnes. Conclusions : These results indicate that Yeouigeumhwang-san has anti-inflammatory effets. If further study is performed, the use of Yeouigeumhwang-san will be valuable and benificial in the therapy of acnes.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.2 s.33
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• pp.116-131
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• 2007

황기건중탕은 보비익기(補裨益氣) 및 거습열(祛濕熱)하는 효능(效能)으로 신체(身體)의 허약(虛弱)을 개선하며 염증(炎症)을 유발할 수 있는 외사(外邪)의 침입을 방어하여 인체의 면역기능(免疫機能)을 강화하는 처방으로 면역관련 세포중 하나인 microglial cell를 대상으로 면역세포에 황기건중탕이 어떠한 효능을 발휘하는 지를 살펴보았다. 물로 전탕(煎湯)한 황기건중탕을 여과한 후 실험한 결과 BV2 microglial cells에서 LPS로 자극하여 염증을 유발하는 주요 물질인 NO의 분비를 세포독성 없이 억제하였다. 또한NO를 생성하는 효소인 Cox-2의 발현도 감소시켰다. 그리고 기타 염증전구 물질들이 $TNF-{\alpha}$, $IL-1{\beta}$, IL-12도 황기건중탕의 용량이 증가함에 따라 의존적으로 감소함을 알 수 있었다. 이러한 결과를 보아 황기건중탕은 면역관련세포인 microglial cell에서 염증 관련 인자들의 분비 및 생성을 억제를 통하여 면역관련 상태를 개선시키는 것으로 사료된다.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.29 no.3
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• pp.74-94
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• 2016

Objectives : YangHyulEum Gami-Bang(YHEG) is a hair care extracts which is composed of fourteen plant extracts used in oriental medicine. The purpose of this study is to investigate the effect of YangHyulEum Gami-Bang(YHEG) on the alopecia and hair growth.Methods & Results : The herbal extracts from YangHyulEum Gami-Bang(YHEG) was tested using in vivo and in vitro test models. 1. The YHEG extracts showed effect on the DNA proliferation of the hair dermal papilla cells measured by [3H]thymidine incorporation. 2. YHEG showed promoting on the expression of growth factors such as IGF-1, KGF-1 and inhibiting on the expression of inhibitory hair growth factor such as TGF-β1, BMP-2 estimated by qPCR. 3. The YHEG extracts showed effect on the activation of β-catenin in the dermal papilla cells. 4. YHEG showed inhibitory effects of NO synthesis at 0.2% concentrations. 5. YHEG showed effects in the expression of IL-1β, TNF-α, IL-6, COX-2 and iNOS gene in the LPS stimulated RAW 264.7 cells. 6. The hair growth index of the YHEG extracts ranked at over 2 when compared to control group which was ranked at 0. 7. The hair follicle number, length and size of the experimental group were remarkably higher than the control group in the histological observation.Conclusions : These results suggest that YangHyulEum Gami-Bang(YHEG) has hair growth promoting activity and it can be used as a potent treatment agent for preventing hair loss and stimulating hair growth for treatment of alopecia.

• The Journal of Internal Korean Medicine
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• v.34 no.2
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• pp.204-214
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• 2013

Objectives : This study was designed to investigate the cellular and molecular mechanisms of anti-inflammatory activity of the water extract of Polygala tenuifolia Willd. (Pt-WE). Methods : Using lipopolysaccharide (LPS)-stimulated murine RAW264.7 cells, we examined inflammatory mediators such as nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$). Also, the inhibitory effect of Pt-WE on the activity of activator protein 1 (AP-1) and upstream signaling molecules was evaluated. To assess the protective effect of Pt-WE on hydrochloride/ethanol (HCl/EtOH)-induced gastric ulcer in mice, we compared Pt-WE (200 mg/kg) with ranitidine (50 mg/kg) treated mice's gastric mucosa, based on gross observations. Results : Pt-WE inhibited LPS-induced production of NO, $PGE_2$ in a dose-dependent manner, without causing cytotoxicity. Pt-WE suppressed AP-1 activation by reducing generations of both c-Jun and c-Fos. In addition, Pt-WE inhibited the p-MKK 4/7 (mitogen-activated protein kinase kinase 4/7) and p-JNK (c-Jun N-terminal kinase) 1 in LPS-stimulated RAW264.7 cells. HCl/EtOH-induced gastric ulcer lesions were inhibited by pre-treatment of Pt-WE based on gross observations. In addition, Pt-WE decreased the phosphorylation level of JNK. Conclusions : These results demonstrate that Pt-WE has anti-inflammatory and gastroprotective effects. Thus, Pt-WE may be used widely in treatment of not only neurodegenerative diseases but also inflammatory diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.31 no.6
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• pp.313-327
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• 2017

The purpose of this study is to predict the effects of macroscopic and integrative therapies by finding active ingredients, potential targets of Astragalus membranaceus (Am) and Cornus officinalis (Co) for diabetic nephropathy. We have constructed network pharmacology-based systematic and network methodology by system biology, chemical structure, chemogenomics. We found several active ingredients of Astragalus membranaceus (Am) and Cornus officinalis (Co) that were speculated to bind to specific receptors which had been known to have a role in the progression of diabetic nephropathy. Four components of Am and eleven components of Co could bind to iNOS; two ingredients of Am and six ingredients of Co could docking to cGB-PDE; one component of Am and nine components of Co could bind to ACE; three ingredients of Co with neprilysin; three components of Co with ET-1 receptor; four ingredients of Am and fourteen ingredients of Co with mineralocorticoid receptor; one component of Am and seven components of Co with interstitial collagenase; one ingredient of Am and ten ingredients of Co with membrane primary amine oxidase; one component of Am and four components of Co with JAK2; two ingredients of Am and one ingredient of Co with MAPK 12; one component of Am and five components of Co could docking to TGF-beta receptor type-1. From this work we could speculate that the possible mechanisms of Am and Co for diabetic nephropathy are anti-inflammatory, antioxidant and antihypertensive effects.