• 대한본초학회지
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• 제32권2호
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• pp.77-85
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• 2017

Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.

• 대한본초학회지
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• 제32권2호
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• pp.107-114
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• 2017

Objectives : Tribulus terrestris $Linn{\acute{e}}$ (Tribuli Fructus; TF) has been used to treat hypochondrium, agalactia, nebula, itching and vitiligo in traditional Korean medicine. In this study, we investigated the effects of TF 30% ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : TF extract was prepared by 30% ethanol. RBL-2H3 cells, a rat mast cell line, were treated with TF extract at different concentrations for 1 hr and then stimulated with DNP-IgE/HSA for indicated times. Cell viability was measured by WST-1 assay. The expression of inflammatory cytokines (IL-4, IL-13 and $IFN-{\gamma}$) mRNA was determined by reverse transcriptase-PCR, and the phosphorylation of ERK1/2, p38 and JNK MAP kinases (MAPKs) was determined by Western blot. The nuclear expression of $NF-{\kappa}B$ p65 in the cells was detected by Western blot and immunocytochemistry, respectively. Results : The treatment of TF extract at 0.1 and $0.2mg/m{\ell}$ significantly decreased the expression of IL-4 and IL-13 mRNA in IgE-stimulated RBL-2H3 mast cells, while significantly increased the expression of $IFN-{\gamma}$ mRNA. TF extract treatment was also inhibited the phosphorylation of ERK1/2, p38 and JNK MAPKs in IgE-stimulated RBL-2H3 mast cells in a dose-dependent manner. In addition, TF extract significantly blocked the translocation of $NF-{\kappa}B$ p65 into the nuclear of cells after IgE stimulation. Conclusions : These results indicate that TF extract inhibits inflammatory response in IgE-stimulated mast cells through blocking MAPKs/$NF-{\kappa}B$ pathway. This suggests that TF extract has an anti-inflammatory activity in mast cell activation.

• 대한본초학회지
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• 제33권2호
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• pp.19-28
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• 2018

Objectives : Epimedii Herba has been frequently used in Korean Traditional Medicine to treat impotence, spermatorrhoea, exophthalmos, and forgetfulness. Present study investigated anti-inflammatory effects of Epimedii Herba water extract (EWE) and attempted to elucidate molecular mechanisms involved. Methods : To explore anti-inflammatory effects of EWE, RAW 264.7 cells, a murine macrophage cell line, were pretreated with $10-100{\mu}g/m{\ell}$ of EWE, and then subsequently exposed to $1{\mu}g/m{\ell}$ of lipopolysaccharide (LPS). Levels of nitric oxide (NO), interleukin-6, $interleukin-1{\beta}$, and tumor necrosis $factor-{\alpha}$ were monitored in the medium. Expression levels of inducible nitric oxide synthase and cyclooxygenase-2 were determined by immunoblot and real-time PCR analyses. Signaling pathways related with nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinases were monitored to elucidate molecular mechanisms involved. Finally, the role of three flavonoid compounds in EWE on LPS-mediated NO production were investigated. Results : In conditioned medium, pretreatment of EWE ($100{\mu}g/m{\ell}$) significantly inhibited LPS-stimulated NO and pro-inflammatory cytokine production. In addition, EWE attenuated the expressions of inducible nitric oxide synthase and cyclooxygenase-2 by LPS. EWE prevented the phosphorylation and degradation of inhibitory ${\kappa}B{\alpha}$, nuclear translocation of $NF-{\kappa}B$, and DNA binding of $NF-{\kappa}B$, while EWE did not change the phosphorylation of mitogen-activated protein kinases by LPS. Moreover, icariin, icaritin, and quercetin partly, but significantly, inhibited the LPS-stimulated NO production. Conclusions : These results suggest that EWE has an ability to prevent inflammation in macrophages through inhibition of $NF-{\kappa}B$ signaling pathway.

• Korean Journal of Acupuncture
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• 제39권2호
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• pp.43-53
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• 2022

Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.

• BMB Reports
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• 제55권2호
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• pp.98-103
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• 2022

Increased mRNA levels of cancer upregulated gene (CUG)2 have been detected in many different tumor tissues using Affymetrix microarray. Oncogenic capability of the CUG2 gene has been further reported. However, the mechanism by which CUG2 overexpression promotes cancer stem cell (CSC)-like phenotypes remains unknown. With recent studies showing that pyruvate kinase muscle 2 (PKM2) is overexpressed in clinical tissues from gastric, lung, and cervical cancer patients, we hypothesized that PKM2 might play an important role in CSC-like phenotypes caused by CUG2 overexpression. The present study revealed that PKM2 protein levels and translocation of PKM2 into the nucleus were enhanced in CUG2-overexpressing lung carcinoma A549 and immortalized bronchial BEAS-2B cells than in control cells. Expression levels of c-Myc, CyclinD1, and PKM2 were increased in CUG2-overexpressing cells than in control cells. Furthermore, EGFR and ERK inhibitors as well as suppression of Yap1 and NEK2 expression reduced PKM2 protein levels. Interestingly, knockdown of β-catenin expression failed to reduce PKM2 protein levels. Furthermore, reduction of PKM2 expression with its siRNA hindered CSC-like phenotypes such as faster wound healing, aggressive transwell migration, and increased size/number of sphere formation. The introduction of mutant S37A PKM2-green fluorescence protein (GFP) into cells without ability to move to the nucleus did not confer CSC-like phenotypes, whereas forced expression of wild-type PKM2 promoted such phenotypes. Overall, CUG2-induced increase in the expression of nuclear PKM2 contributes to CSC-like phenotypes by upregulating c-Myc and CyclinD1 as a co-activator.

• Journal of Ginseng Research
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• 제47권1호
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• pp.106-116
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• 2023

Background: Pirarubicin (THP) is an anthracycline antibiotic used to treat various malignancies in humans. The clinical usefulness of THP is unfortunately limited by its dose-related cardiotoxicity. Ginsenoside F1 (GF1) is a metabolite formed when the ginsenosides Re and Rg1 are hydrolyzed. However, the protective effects and underlying mechanisms of GF1 on THP-induced cardiotoxicity remain unclear. Methods: We investigated the anti-apoptotic and anti-oxidative stress effects of GF1 on an in vitro model, using H9c2 cells stimulated by THP, plus trigonelline or AKT inhibitor imidazoquinoxaline (IMQ), as well as an in vivo model using THP-induced cardiotoxicity in rats. Using an enzyme-linked immunosorbent test, the levels of malondialdehyde (MDA), brain natriuretic peptide (BNP), creatine kinase (CK-MB), cardiac troponin (c-TnT), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) were determined. Nuclear factor (erythroid-derived2)-like 2 (Nrf2) and the expression of Nrf2 target genes, including heme oxygenase-1 (HO-1), glutathione-S-transferase (Gst), glutamate-cysteine ligase modifier subunit (GCLM), and expression levels of AKT/Bcl-2 signaling pathway proteins were detected using Western blot analysis. Results: THP-induced myocardial histopathological damage, electrocardiogram (ECG) abnormalities, and cardiac dysfunction were reduced in vivo by GF1. GF1 also decreased MDA, BNP, CK-MB, c-TnT, and LDH levels in the serum, while raising SOD and GSH levels. GF1 boosted Nrf2 nuclear translocation and Nrf2 target gene expression, including HO-1, Gst, and GCLM. Furthermore, GF1 regulated apoptosis by activating AKT/Bcl-2 signaling pathways. Employing Nrf2 inhibitor trigonelline and AKT inhibitor IMQ revealed that GF1 lacked antioxidant and anti-apoptotic effects. Conclusion: In conclusion, GF1 was found to alleviate THP-induced cardiotoxicity via modulating Nrf2 and AKT/Bcl-2 signaling pathways, ultimately alleviating myocardial oxidative stress and apoptosis.

• 생명과학회지
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• 제33권2호
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• pp.176-182
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• 2023

석곡의 줄기는 전통 동양의학에서 위를 보하고, 진액을 보충하며, 열을 내리는 것에 사용되어 왔다. 본 연구에서는 RBL-2H3 세포에서 비만세포 탈과립과 TNF-α, IL-4, histidine decarboxylase (HDC) 발현에 미치는 석곡 열수추출물(DME)의 효과를 조사하였다. DME는 PMA와 Calcium ionophore 병행처리(PMACI)에 의해 유도되는 β-hexosaminidase 분비와 TNF-α, IL-4, HDC 발현을 현저히 억제하였다. 또한 PMACI에 의해 유도되는 NF-κB, AP-1 활성과 p38 kinase, extracellular signal-regulated kinase 1/2 (ERK1/2)과 c-Jun N-terminal kinase (JNK)의 인산화가 DME 전처리에 의해 저해되었다. 이러한 결과들은DME가 비만세포 탈과립을 억제하고, MAPKs/NF-κB/AP-1 신호전달 경로를 통해 TNF-α, IL-4, HDC 발현을 억제한다는 것을 시사한다. 본 연구결과들로 보아 DME는 과민반응과 염증성 질환을 치료하는 약물로 개발될 가능성을 가지는 것으로 사료된다.

• 한국작물학회지
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• 제31권2호
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• pp.195-203
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• 1986

도체내 무기양분이 등숙진전에 따라 이행 집적되는 정도와 주이동부위에 대한 검토를 위하여 생태형과 숙기가 상이한 4품종을 재배하여 출수기부터 1주간격으로 9회에 걸쳐 시료를 탐취 분석한 결과는 다음과 같다. 1. 인산은 등숙진전에 따라 현미내 함유율이 증가되고 경엽에서는 감소하여 인산의 재이동을 인정할수가 있고 이동정도는 절간에서 가장 많고 다음이 엽초, 엽신의 순이었으며 이삭에서 가까운 상위의 절간, 엽초, 엽신에서의 이동이 하위에서 보다 컸다. 생태형간에 차가 있고 통일형에서 이동이 많은 것으로 나타났다. 2. 가리는 엽신과 엽초에서는 등숙진전에 따른 변동이 없는 반면 절간에서는 현저히 증가되었고 특히상위절간에 집적이 많았다. 현미에서는 성숙기민 6주까지 함유율이 점진적으로 감소하다가 7주부터 다시 증가하여 절간과 반대경향을 보였으나 기간중 현미중의 증가를 고찰하면 단위미립내의 가리함양의 변동은 미미하여 절간의 가리축적이 현미에서 재이동된 것으로 볼 수는 없었다. 도체내 부위별 가리함양은 품종의 생태형보다 숙기에 따른 차가 큰 것으로 나타났다. 3. 고토는 등숙이 진전됨에 따라 해미내 함유율이 점진적으로 증가되고 경엽에서는 호숙기까지 증가되나 이후 성숙기까지 감소경향을 보여 인산만은 못해도 곡실로의 이동을 인정할 수가 있었고 이동상태는 부위나 숙기, 생태형에 따른 경향은 인정하기 어려웠고 품종간에는 차가 있었다. 4. 규산은 현미에서 미량이 검출될 뿐이나 왕겨에서는 등숙진전에 따라 계계 단가되고 경엽에서도 증가되나 엽신>엽초>절간의 순으로 축적되어 재이동은 생각할 수가 없었고 품종의 생태형간의 는 발견할 수가 없었으나 숙기간에는 차가 있어 조생종보다 중생종인 상풍벼와 금강벼의 경엽에 함유율이 높았다. 5. 석회는 곡실로의 이동이나 축적은 없었고 엽초와 절간에서도 변동이 없었으나 엽신에서는 등숙진전에 따라 축적됨을 알 수 있었고 특히 다수계 품종민 태백벼와 금강벼에서 축적정도가 컸다. 엽위별로는 지금이 많고 차엽이 낮았다.

• 문화기술의 융합
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• 제9권1호
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• pp.717-724
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• 2023

사플라워와 마더워트 등 에탄올복합추출물(SEC)을 lipopolysaccharide(LPS)로 유도된 마우스 대식세포주인 RAW264.7 세포와 마우스 폐포 대식세포주인 MH-S 세포에서 사플라워와 마더워트 등 에탄올복합추출물((SEC)의 항균과 항염증 활성 효과를 조사하였다. 그 결과 SEC(500 ㎍/mL)의 전처리 LPS 자극 세포에서 iNOS 단백질 및 염증성 사이토카인 mRNA 발현을 크게 억제하였다. 사플라워와 마더워트 등 에탄올복합추출물(SEC)에 의한 항염증활성 효과는 다음과 같이 관찰되었다. 사플라워와 마더워트 등 에탄올복합추출물(SEC)은 IκB-α 인산화의 억제를 통해 시토졸에서 핵으로의 NF-κB의 전좌를 억제하고 또한 LPS로 자극된 NF-κB 전사 활성을 억제하였다. 이러한 결과는 사플라워와 마더워트 등 에탄올복합추출물(SEC)이 항염증 작용을 발휘 하고 사플라워와 마더워트 등 에탄올복합추출물(SEC)의 잠재적인 치료 가치성 및 기본 메커니즘을 규명 할 수 있었다. 결론적으로 사플라워와 마더워트 등 에탄올복합추출물(GRP)이 다양한 항염증 활성성분 함유 및 천연 항염증제의 잠재적 공급원으로 활용 가능성이 있음을 시사 하였다.

• Journal of Ginseng Research
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• 제47권3호
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• pp.458-468
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• 2023

Background: As a complication of Type II Diabetes Mellitus (T2DM), the etiology, pathogenesis, and treatment of cognitive dysfunction are still undefined. Recent studies demonstrated that Ginsenoside Rg1 (Rg1) has promising neuroprotective properties, but the effect and mechanism in diabetes-associated cognitive dysfunction (DACD) deserve further investigation. Methods: After establishing the T2DM model with a high-fat diet and STZ intraperitoneal injection, Rg1 was given for 8 weeks. The behavior alterations and neuronal lesions were judged using the open field test (OFT) and Morris water maze (MWM), as well as HE and Nissl staining. The protein or mRNA changes of NOX2, p-PLC, TRPC6, CN, NFAT1, APP, BACE1, NCSTN, and Ab1-42 were investigated by immunoblot, immunofluorescence or qPCR. Commercial kits were used to evaluate the levels of IP3, DAG, and calcium ion (Ca²⁺) in brain tissues. Results: Rg1 therapy improved memory impairment and neuronal injury, decreased ROS, IP3, and DAG levels to revert Ca²⁺ overload, downregulated the expressions of p-PLC, TRPC6, CN, and NFAT1 nuclear translocation, and alleviated Aβ deposition in T2DM mice. In addition, Rg1 therapy elevated the expression of PSD95 and SYN in T2DM mice, which in turn improved synaptic dysfunction. Conclusions: Rg1 therapy may improve neuronal injury and DACD via mediating PLC-CN-NFAT1 signal pathway to reduce Aβ generation in T2DM mice.