• 한방안이비인후피부과학회지
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• 제34권3호
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• pp.1-12
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• 2021

Objectives : The purpose of this study is to investigate the anti-inflammatory effect of Geumeunwha-san(GEHS) water extract in vitro. Methods : To evaluate the anti-inflammatory effect of GEHS, Raw 264.7 cells were pretreated with 10-300㎍/㎖ of GEHS for 1hr, and then exposed to 1㎍/㎖ of LPS. MTT assay was used to detect the cell viability. Productions of pro-inflammatory cytokines and NO were measured in culture media. By using immunoblot analysis, protein levels of iNOS and NF-𝜅B were determined. Results : In vitro study, cell viability assay on GEHS treatment of 10-300㎍/㎖ has no cytotoxicity in Raw 264.7 cells. Pretreated 100, 300㎍/㎖ of GEHS had significantly inhibited LPS-induced NO production. And also pretreatment of 100, 300㎍/㎖ GEHS had significantly decreased production of interleukin-6, -1𝛽 and tumor necrosis factor-𝛼 in LPS-activated Raw 264.7 cells. In addition, GEHS reduced LPS-mediated iNOS expression. Moreover I-𝜅B𝛼 expression was significantly induced by GEHS and NF-𝜅B expression was reduced by GEHS. Conclusions : These results suggest the clinical basis of GEHS for the treatment of inflammatory diseases.

• The Korean Journal of Pain
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• 제34권2호
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• pp.176-184
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• 2021

Background: Diabetes-related neuropathic pain frequently occurs, and the underpinning mechanism remains elusive. The periaqueductal gray (PAG) exhibits descending inhibitory effects on central pain transmission. The current work aimed to examine whether inflammatory cytokines regulate mechanical allodynia and thermal hyperalgesia induced by diabetes through the phosphoinositide 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) pathway in the PAG. Methods: Streptozotocin (STZ) was administered intraperitoneally to mimic allodynia and hyperalgesia evoked by diabetes in rats. Behavioral assays were carried out for determining mechanical pain and thermal hypersensitivity. Immunoblot and ELISA were performed to examine PAG protein amounts of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), as well as their corresponding receptors in STZ rats, and the expression of PI3K/protein kinase B (Akt)/mTOR signaling effectors. Results: Increased PAG p-PI3K/p-Akt/p-mTOR protein amounts were observed in STZ-induced animals, a PI3K-mTOR pathway inhibition in the PAG attenuated neuropathic pain responses. Moreover, the PAG concentrations of IL-1β, IL-6, and TNF-α and their receptors (namely, IL-1R, IL-6R, and tumor necrosis factor receptor [TNFR] subtype TNFR1, respectively) were increased in the STZ rats. Additionally, inhibiting IL-1R, IL-6R, and TNFR1 ameliorated mechanical allodynia and thermal hyperalgesia in STZ rats, alongside the downregulation of PI3K-mTOR signaling. Conclusions: Overall, the current study suggests that upregulated proinflammatory cytokines and their receptors in the PAG activate PI3K-mTOR signaling, thereby producing a de-inhibition effect on descending pathways in modulating pain transmission, and eventually contributing to neuropathic pain.

• Journal of Ginseng Research
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• 제46권4호
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• pp.572-584
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• 2022

Background: Huntington's disease (HD) is a neurodegenerative disorder caused by the expansion of trinucleotide CAG repeat in the Huntingtin (Htt) gene. The major pathogenic pathways underlying HD involve the impairment of cellular energy homeostasis and DNA damage in the brain. The protein kinase ataxia-telangiectasia mutated (ATM) is an important regulator of the DNA damage response. ATM is involved in the phosphorylation of AMP-activated protein kinase (AMPK), suggesting that AMPK plays a critical role in response to DNA damage. Herein, we demonstrated that expression of polyQ-expanded mutant Htt (mHtt) enhanced the phosphorylation of ATM. Ginsenoside is the main and most effective component of Panax ginseng. However, the protective effect of a ginsenoside (compound K, CK) in HD remains unclear and warrants further investigation. Methods: This study used the R6/2 transgenic mouse model of HD and performed behavioral tests, survival rate, histological analyses, and immunoblot assays. Results: The systematic administration of CK into R6/2 mice suppressed the activation of ATM/AMPK and reduced neuronal toxicity and mHTT aggregation. Most importantly, CK increased neuronal density and lifespan and improved motor dysfunction in R6/2 mice. Conversely, CK enhanced the expression of Bcl2 protected striatal cells from the toxicity induced by the overactivation of mHtt and AMPK. Conclusions: Thus, the oral administration of CK reduced the disease progression and markedly enhanced lifespan in the transgenic mouse model (R6/2) of HD.

• BMB Reports
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• 제55권11호
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• pp.571-576
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• 2022

Advancements in the field of proteomics have provided opportunities to develop diagnostic and therapeutic strategies against various diseases. About half of the world's population remains at risk of malaria. Caused by protozoan parasites of the genus Plasmodium, malaria is one of the oldest and largest risk factors responsible for the global burden of infectious diseases with an estimated 3.2 billion persons at risk of infection. For epidemiological surveillance and appropriate treatment of individuals infected with Plasmodium spp., timely detection is critical. In this study, we used combinations of depletion of abundant plasma proteins, 2-dimensional gel electrophoresis (2-DE), image analysis, LC-MS/MS and western blot analysis on the plasma of healthy donors (100 individuals) and vivax and falciparum malaria patients (100 vivax malaria patients and 8 falciparum malaria patients). These analyses revealed that α1-antichymotrypsin (AACT) protein levels were elevated in vivax malaria patient plasma samples (mean fold-change ± standard error: 2.83 ± 0.11, based on band intensities), but not in plasma from patients with other mosquito-borne infectious diseases. The results of AACT immunoblot analyses showed that AACT protein was significantly elevated in vivax and falciparum malaria patient plasma samples (≥ 2-fold) compared to healthy control donor plasma samples, which has not been previously reported.

• 대한본초학회지
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• 제33권4호
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• pp.35-41
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• 2018

Objectives : Hyeonggaeyeongyotang Gagambang (HYT) is a herbal medicine prescribed for the treatment of inflammatory diseases, but it is necessary to study the exact therapeutic efficacy. This study aims to investigate the antibacterial and anti-inflmmatory activities of HYT. Methods : Antibacterial activity of HYT was confirmed by staining Escherichia coli, a gram negative strain, and Staphylococcus aureus, a gram positive strain, on solid Lysogeny Broth (LB) medium containing HYT. Antioxidant activity of HYT was confirmed by 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. The phosphorylation of nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha ($I{\kappa}B{\alpha}$) after lipopolysaccharide (LPS) treatment with HYT-treated RAW 264.7 mouse macrophages cells was confirmed by immunoblot analysis and the level of interleukin 1 beta (IL-$1{\beta}$) mRNA expression level was confirmed by quantitative real-time PCR. Results : HYT showed a concentration-dependent antibacterial activity against Escherichia coli and Staphylococcus aureus and also showed excellent antioxidant activity. HYT treatment attenuated the phosphorylation of $I{\kappa}B{\alpha}$induced by LPS treatment in RAW 264.7 mouse macrophages cells. The phosphorylation of $I{\kappa}B{\alpha}$is crucial for the regulation of the expression of various pro-inflammatory cytokines. In addition, IL-$1{\beta}$ mRNA expression level of RAW 264.7 mouse macrophages cells stimulated by LPS treatment was also inhibited by HYT treatment. Conclusions : Through experimental demonstration of the antioxidative, antimicrobial and anti-inflammatory effects of HYT, we demonstrated that HYT is a herbal medicine effective for the treatment of inflammatory diseases caused by various bacterial infections.

• 대한본초학회지
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• 제33권5호
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• pp.9-18
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• 2018

Objectives : This study was performed to determine the transdermal effects of ethanol extract from medicinal herbal mixture (SHJ) on hair growth in C57BL/6 mice and melanogenesis in melanoma cells. Methods : Mice were divided into 3 experimental groups including vehicle (CON), SHJ extract and 5% minoxidil (MNXD, positive control)-treated group. SHJ was applied topically on the hair-shaved skin of C57BL/6 mice everyday for 15 days. The thickness and density of hair with a folliscope and morphometry of hair follicle with a H&E staining were monitored at last day. Also then, hair growth-associated gene expressions were measured by immunoblot assay. Results : The MNXD or SHJ-treated group promoted on hair growth compared to that of vehicle-treated group (CON). Hair density and thickness of MNXD or SHJ treated-group increased compared to that of vehicle application on the 15 days, respectively. Induction of insulin-like growth factor (IGF)-1 and vascular endothelial growth factor (VEGF) were also accelerated by application of SHJ extract compared to those of CON group. But expression of transforming growth factor (TGF)-${\beta}1$ decreased in SHJ treated-group compared to that of CON group. Furthermore, SHJ extract showed to increase melanin contents in a dose-dependent manner. Tyrosinase activity significantly increased in SHJ-treated group compared with CON group in dose-dependant manner. Conclusions : These results suggest that SHJ can be used as a component of cosmeceuticals for hair care via promoting growth and melanogenesis of hair.

• 대한본초학회지
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• 제34권2호
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• pp.59-66
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• 2019

Objectives : Decrease in muscle mass and loss of muscle function due to aging are associated with various diseases. As interest in healthy aging increases, efforts to prevent and treat muscle hypoxia as an illness are increasing. Considering the physical limitations, a pharmacologic approach to the treatment of myopenia is needed. Methods : Terminalia chebula Rets has a wide range of pharmacological effects and is used as a medicinal product in traditional medicine. However, the drug effect on the treatment of muscle disorders has not been revealed. The purpose of this study was to evaluate the value of water extract of Terminalia chebula (WETC) as a therapeutic agent to relieve symptoms of muscle hypoxia. Results : WETC showed strong radical scavenging ability. In addition, WETC increased cell activity of myoblast, and we observed that WETC induces myoblast differentiation by immunoblot analysis using differentiation protein markers as well as cell morphology of myoblast. Based on these results, we examined the effect of chebulic acid, chebulagic acid, gallic acid, geraniin, and punicalagin on cell activity and differentiation of myoblasts. Gallic acid significantly increased cell activity of myoblast, and it was found to be an effective substance which not only induces myoblast differentiation but also promotes proliferation. Conclusions : We suggest that the WETC with antioxidant effect and its indicator gallic acid on cell activity, proliferation and differentiation of myoblast can be studied and developed as a food and medicine for prevention and treatment of various muscle diseases.

• 대한본초학회지
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• 제35권3호
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• pp.9-16
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• 2020

Objectives : Sarcopenia is a disease that leads to a decrease in skeletal muscle, and the importance of prevention and treatment thereof is increasing in an aging society. However, there is a definite limitation of exercise therapy for sarcopenia, and thus, there is an urgent need for a pharmacologic research to the treatment of sarcopenia. Methods : 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay was performed to confirm the antioxidant efficacy of water extract of Lacca Sinica Exsiccata (WELSE). To determine the effect of WELSE on myoblast activity, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed. To confirm the effect of WELSE on the differentiation of myoblast into myotubes, protein expression levels of myosin heavy chain 3 (Myh3) and paired box 3/7 (pax3/7) were confirmed by immunoblot analysis. In addition, immunofluorescence microscopy was performed to confirm the effect on myotube formation of WELSE. Results : It was confirmed that WELSE had high antioxidant activity and showed no cytotoxicity to myoblast up to 200 ㎍/㎖ concentration. Myoblast was treated with WELSE at a concentration of 100 ㎍/㎖ and differentiated for 5 days. The expression of Myh3, which forms myotubes, was promoted and the morphology of myotubes was changed and Increasing the thickness. Conclusions : In this paper, we confirmed the excellent antioxidant efficacy of WELSE and positive effects on muscle differentiation and myotube formation. These results suggest valuable as a material for pharmaceutical research on the prevention and treatment of sarcopenia.

• Parasites, Hosts and Diseases
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• 제60권1호
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• pp.25-34
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• 2022

Alveolar echinococcosis (AE) caused by infection with E. multilocularis metacestode, represents one of the most fatal helminthic diseases. AE is principally manifested with infiltrative, proliferating hepatic mass, resembling primary hepatocellular carcinoma. Sometimes metastatic lesions are found in nearby or remote tissue. AE diagnosis largely depends on imaging studies, but atypical findings of imaging features frequently require differential diagnosis from other hepatic lesions. Serological tests may provide further evidence, while obtaining reliable AE materials is not easy. In this study, alternative antigens, specific to AE were identified by analyzing E. granulosus protoscolex proteins. An immunoblot analysis of E. granulosus protoscolex showed that a group of low-molecular-weight proteins in the range from 14 kDa to 16 kDa exhibited a sensitive and specific immune response to AE patient sera. Partial purification and proteomic analysis indicated that this protein group contained myosin, tubulin polymerization promoting protein, fatty-acid binding protein, uncharacterized DM9, heat shock protein 90 cochaperone tebp P-23, and antigen S. When the serological applicability of recombinant forms of these proteins was assessed using enzyme-linked immunosorbent assay, DM9 protein (rEgDM9) showed 90.1% sensitivity (73/81 sera tested) and 94.5% specificity (172/181 sera tested), respectively. rEgDM9 showed weak cross-reactions with patient sera from the transitional and chronic stages of cystic echinococcosis (3 to 5 stages). rEgDM9 would serve as a useful alternative antigen for serodiagnosis of both early- and advanced-stage AE cases.

• 대한본초학회지
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• 제24권4호
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• pp.39-47
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• 2009

Objectives : Hwangnyeonhaedok-tang (Huanglian Jiedu Tang; HHT) has been widely used for purging' 'fire' and lessening virulence of any pathogenic organism. However it has been rarely conducted to evaluate the immuno-biological activity. In this study, we evaluated anti-inflammatory effects of HHT in LPS-activated Raw264.7 cells. Methods : Cells were treated with $1\;{\mu}g/ml$ of LPS 1 h prior to the addition of HHT. Cell viability was measured by MTT assay. The production of NO was determined by reacting cultured medium with Griess reagent. PGE2 and proinflammatory cytokines were detected by ELISA. Expression of iNOS, COX-2, $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ were analyzed by immunoblot analysis. Results : All three doses of HHT (0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity during the entire experimental period. The levels of NO and PGE2 were dramatically augmented by LPS compared to control. However, HHT extract dose-dependently reduced these increases. Expression of iNOS and COX-2 protein were also decreased by treatment with HHT extract. Furthermore, HHT extract significantly reduced the nuclear translocation of NF-${\kappa}B$ which is critical in regulating inflammation through transcription of iNOS and COX-2. In addition, HHT extract reduced the elevated production of inflammatory cytokines including TNF-$\alpha$, IL-$1{\beta}$ and IL-6. Conclusions : The results in this study demonstrate that HHT extract exerts anti-inflammatory activities through the inhibition of NO, PGE2 and proinflammatory cytokines production via the suppression of NF-${\kappa}B$.