• 대한한방부인과학회지
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• 제36권3호
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• pp.1-24
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• 2023

Objectives: Boeum-jeon (BEJ) is a herbal formula composed 8 Korean medicinal herbs and is traditionally used to treat inflammatory diseases. In this study, the authors tried to confirm the antioxidant efficacy of BEJ and its anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Methods: In this experiment, results of BEJ on the following two were measured as follows: (1) Antioxidant effects was measured by DPPH Radical scavenging Activity, ABTS Radical scavenging Activity. (2) Anti-Inflammatory effects were evaluated by the production amount of ROS, NO, Cytokine (PGE2, IL-1β, IL-6, TNF-α), COX-2, iNOS, TNF-α, IL-1β, IL-6, HO-1, NQO1 (the previous seven are "mRNA Expression"), COX-2, iNOS, TNF-α, IL-1β, IL-6, HO-1, NQO1, NRF2 (the previous eight are "Protein Expression") ERK, JNK, p38 (the previous three are "Protein Phosphorylation") in LPS-stimulated RAW 264.7 cells. Results: The experimental measurement results are as follows: (1) DPPH Radical scavenging Activity, ABTS Radical scavenging Activity increased in a concentration -dependent manner in the BEJ-treated group. (2) As a result of measuring anti-inflammatory efficacy, the production of ROS, NO, and Cytokine (PGE2, TNF-α, IL-1β, IL-6) in the BEJ-administered group was significantly reduced compared to the control group. (3) Among mRNA Expression levels, COX-2, iNOS, IL-1β, IL-6 and TNF-α was significantly decreased in a concentration-dependent manner than in the control group, and HO-1 and NQO1 were significantly increased in a concentration -dependent manner than in the control group. (4) Among the Protein Expression levels, COX-2, iNOS, IL-1β, IL-6 and TNF-α was significantly decreased in a concentration -dependent manner compared to the control group, and HO-1, NQO1 and NRF2 was significantly increased in a concentration-dependent manner compared to the control group. (5) As a result of Protein Phosphorylation, ERK, p38 and JNK was significantly decreased compared to the control group in a concentration-dependent manner. Conclusions: Boeum-jeon has been experimentally confirmed to have antioxidant and anti-inflammatory effects, and if the evidence for efficacy is reinforced through further studies such as in vivo studies and clinical trials in the future, it can be effectively used to treat various inflammatory diseases such as bladder inflammation and chronic pelvic inflammation.

• Journal of Chest Surgery
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• 제35권6호
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• pp.407-419
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• 2002

죽상경화증은 지방, 대식세포나 평활근세포와 같은 세포, 그리고 extracellular matrix(ECM)의 점진적인 축적이 특징적인 만성 염증성 혈관 질환이다. Matrix metalloprotenases(MMPs)와 tissue inhibitor of metalloproteinases(TIMs)는 죽상경화증에서 혈관의 ECM의 분해와 재모델링에 관여하며, cytokines는 MMPs와 TIMPs의 합성이나 활성화에 관여하는 것으로 보고된 바 있다. 대상 및 방법： 연구 대상으로는 체중 2.0~2.5 kg의 생후 1개월 된 뉴질랜드산 수토끼를 선택하였으며, 10 마리는 12주 동안 1% 콜레스테롤 식이를 투여한 후 실험군으로 이용하였으며, 나머지 10마리는 표준 실험실 식이를 먹여 대조군으로 이용하였다. 12주간 사육 후 토끼를 희생시켰으며, 생존한 실험군 9 마리와 대조군 10 마리의 대동맥과 관상동맥에서 H&E 염색, 면역조직화학 염색, immunoblotting, bioassay의 방법으로 MMP-9, TIMP-2, IL-18의 발현 및 IL-6의 생물학적 활성도를 조사하였다. 결과： 실험군의 혈청 콜레스테롤은 1258$\pm$262mg/dL로 대조군의 41$\pm$7mg/dL에 비하여 유의하게 증가하였다. 실험군의 전예에서 대동맥과 관상동맥에 죽상경화반이 잘 형성되었으며, 실험군의 대 동맥 내막의 두께는 0.31$\pm$0.1mm로 대조군의 0.01mm에 비해 유의하게 증가하였다. 죽상경화반에서 실험군의 MMP-9의 발현은 대조군에 비하여 유의하게 증가하였으며, 내막의 파열이나 관상동맥의 내강 폐쇄가 있었던 증례에서는 더욱 강한 MMP-9의 발현을 관찰할 수 있었다. TIMP-2는 실험군의 일부에서 약한 발현을 보였으나 대조군과 유의한 차이가 없었다. 실험군과 대조군에서 측정한 IL-6의 생물학적 활성도는 각각 4819.60$\pm$2021.25, 27.20$\pm$12.19IU/mL로서 실험군에서 유의한 증가를 보였으며, 면역조직화학 염색에 의한 IL-18의 발현은 대조군에서는 발현되지 않았으나, 실험군은 전예에서 발현을 보였다. 결론： MMP-9의 증가된 발현과 TIMP-2의 무변화로 인한 MMPs/TIMPs의 불균형은 죽상경화성 병변에서 ECM의 분해와 경화반의 불안정화를 촉진시킬 수 있는 것으로 보인다. 또한 내막의 파열이 관찰된 증례에서의 더욱 증가된 MMP-9은 경화반의 파열과 관련있는 것으로 생각된다. IL-6의 생물학적 활성도의 증가 및 IL-18의 발현은, IL-6와 IL-18이 죽상경화증의 표지자일 뿐만 아니라 MMP-9의 분비 또는 활성화에 관여하여 죽상경화증의 진행과 경화반의 불안정성 등에 활발히 참여하는 cytokines임을 시사하는 소견으로 보인다. MMPs, TIMPs, cytokines등의 조절 과정을 밝혀내는 것은 죽상경화증의 세포, 분자적인 병리기전을 이해하는 데에 도움을 줄 것이며, 죽상 경화증의 치료 또는 합병증을 예방할 수 있는 기전을 확립하는 데에 도움이 될 것으로 생각된다.

• 대한한방부인과학회지
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• 제35권3호
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• pp.1-23
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• 2022

Objectives: The purpose of this study is to evaluate anti-breast cancer and anti-inflammatory effects of Chungganhaewool-tang and Shipyeukmiyeugi-eum. Methods: MDA-MB-231 cells were used to measure cytotoxicity, Reactive oxygen species (ROS) production, protein expression amounts of Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), B-cell lymphoma-extra large (Bcl-xl), Cytochrome C Caspase-3, Caspase-7, Caspase-9, Poly ADP-ribose polymerase (PARP), Nuclear factor erythroid-2-related factor 2 (Nrf2), Heme oxygenase-1 (HO-1) and NAD (P) H Quinone Oxidoreductase 1 (NQO1) to evaluate the anti-breast cancer effects of Chungganhaewool-tang (CHT) and Shipyeukmiyeugi-eum (SYE), and THP-1 cells, differentiated into macrophage and induced inflammation with Lipopolysaccharide (LPS), were used to measure production amounts of ROS, Nitric oxide (NO), and protein expression amounts of Inducible nitric oxide synthase (iNOS), Cyclooxygenase (COX-2), Interleukin-1 beta (IL-1β), Interleukin-6 (IL-6) and Tumor necrosis factor-alpha (TNF-α) to evaluate the anti-inflammatory effects of CHT and SYE. Results: CHT and SYE reduced MDA-MB-231 cell counts, increased protein expression of Bax and Cytochrome C, and decreased protein expression of Bcl-2, Bcl-xl. The protein expression amounts of Caspase-3, 7, and 9 decreased, but amounts of the active form, cleaved Caspase-3, 7, and 9, increased. In addition, PARP protein expression decreased, the amount of PARP protein in the cleaved form increased, and the amount of protein expressions of Nrf2 and HO-1 decreased, but NQO1 showed no significant difference. In THP-1 cells CHT and SYE reduced ROS and NO, and reduced protein expressions of iNOS, COX-2, IL-1, and TNF-α, but only SYE groups reduced IL-6. Conclusions: This study suggests that CHT and SYE have potential to be used as treatments for breast cancer.

• 한방안이비인후피부과학회지
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• 제25권1호
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• pp.22-32
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• 2012

Objectives : In this study, Samhwangsaejaegami extract was tested to prove its anti-atopic dermatitis effect on NC/Nga mice. Methods : In order to evaluate the external use effects of Samhwangsaejae extract on anti-atopic dermatitis, the expression of filaggrin, serine palmitoyltransferase(SPT), and COX-2 were analyzed. In vivo study, clinical skin severity score, IgE, IL-4, IL-5 and IL-6 level were analyzed through NC/Nga atopic mice model after 12 weeks external treatment. Results : In vitro study results showed the reduction in the expression of filaggrin, SPT, and COX-2. In vivo study results demonstrated the significant reduction in clinical skin severity score, IgE, IL-4, IL-5, IL-6 expression level. Conclusions : These results showed Samhwangsaejaegami extract can be a promising candidate for anti-atopic dermatitis treatment.

• 한국임상수의학회지
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• 제14권1호
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• pp.30-36
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• 1997

It is well estabilished that IL-4, IL-6, $IFN{\gamma}$ and immunoglobulin production are increased as a consequence of advancing age, and endothelial cells increase their expression of pro-inflammatory surface protein, such as ICAM-1, as a consequence of contact with inflammatory cytokines, including IL-1, $TNF{\alpha} or INF{\gamma}.$ This study was designed to define the relationship between age-associated changes of cytokines and immunoglobulin production and ICAM-1 expression in aged mice. Serum from aged mice had elevated IL-6 and immunoglobulin levels compared to mature adult controls and activation-induced production of $IFN{\gamma}$ of splenocytes from aged mice were increased above normal adult level. By immunoperoxidase staining, ICAM-1 in hearts of normal adult mice was expressed occasionally at low levels, but in aged mice, the number of positive blood vessels for ICAM-1 were increased and also the immunoreactivity were stronger. Therefore, these finding indicate that the altered expression of ICAM-1 eith normal aging may actually be correlative to these age-related changes in cytokiness and immunoglobulin production.

• Journal of Acupuncture Research
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• 제34권1호
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• pp.11-21
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• 2017

Objectives : In this study, we investigated the anti-inflammatory and anti-oxidative effects of MOK, a pharmacopuncture medicine, in lipopolysaccharide (LPS)-stimulated mouse peritoneal macrophages. Methods : Peritoneal macrophages were isolated from ICR mice. Primary macrophages were treated with MOK extract (1.25, 2.5, 5, 10, and 20 mg/ml) for 30 min and then stimulated with LPS ($1{\mu}g/ml$) for the indicated times. Cytotoxicity was measured using MTT and LDH assays. Nitric oxide (NO) production in culture supernatants was measured using the Griess assay. The mRNA expression of iNOS, COX-2, proinflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, and IL-6) and antioxidant enzymes (HO-1 and MnSOD) was measured by RT-PCR. Results : Treatment with MOK extract (2.5, 5, and 10 mg/ml) significantly decreased LPS-induced NO production in peritoneal macrophages through inhibition of iNOS expression. The expression of COX-2, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 mRNA was also decreased in LPS-stimulated macrophages upon treatment with MOK extract. MOK treatment also increased the expression of HO-1 and MnSOD mRNA in macrophages. Conclusion : These results indicate that MOK exerts anti-inflammatory and antioxidant effects by regulating the transcription levels of inflammatory mediators and antioxidant proteins in activated macrophages.

• 대한한방내과학회지
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• 제29권2호
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• pp.334-347
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• 2008

Background and Objective : Haeyeol-tang, composed of Houttuyniae Herba, Lonicerae Flos, Taraxaci Herba, and Scrophulariae Radix, is widely used for alleviating the symptom of various kinds of inflammatory pulmonary disease, including asthma and COPD. We want to know whether Haeyeol-tang has an anti-inflammatory effect by analyzing expression of pro-inflammatory cytokines. Materials and Methods : We differentiated the THP-1 cells into macrophage-like cells by treatment with PMA. Inflammation was induced by treatment with LPS and PMA. We found the safe concentration of Haeyeol-tang by using MTS assay and used PD98059 as a negative control for comparison of anti-inflammatory effect of Haeyeol-tang. Results : The RT-PCR analysis results show that the cell survival rate is over 100% within 1 ng/mL to 1 ug/mL of Haeyeol-tang and begins to decrease under 100% at 10 ug/mL. The gene expression of $IL-1{\beta}$, IL-6, IL-8, IL-10, $TNF-{\alpha}$ and $TGF-{\beta}$ levels were down-regulated when Haeyeol-tang was treated at concentrations between 1 ng/mL an 1 ug/mL on monocyte-derived macrophages. Interestingly, 1 ug/mL Haeyeol-tang-treated samples showed that the transcriptional activities of IL-8, $TNF-{\alpha}$, IL-10 and $TGF-{\beta}$ were more down-regulated than those of PD098059 $(TNF-{\alpha}$ inhibitor). At protein level, the ELISA analysis results showed that there were more remarkable (p<0.001) decreases in expression of $IL-1{\beta}$, IL-6, IL-8 and $TNF-{\alpha}$ on both the 1 ug/mL Haeyeol-tang-treated group and the PD98059-treated group than the LPS-treated group. Conclusion : We conclude that Haeyeol-tang has an anti-inflammatory effect by inhibiting expression of pro-inflammatory cytokines and chemokines at mRNA and protein levels. These results may provide us a promising way to care for general inflammatory diseases as well as inflammatory pulmonary disease, including asthma and COPD, with further clinical study.

• Clinical and Experimental Pediatrics
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• 제52권6호
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• pp.667-673
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• 2009

목 적 : TLR2는 숙주의 항결핵 방어면역의 중요한 역할을 하는 것으로 알려져 있다. 본 연구는 BCG 접종 후 발생한 화농성 림프절염 환자 단핵구에서 TLR2의 발현과 TLR2 리간드 자극에 의한 $TNF-{\alpha}$와 IL-6의 생성을 조사하여 화농성 림프절염 발병과 TLR2의 연관성을 알아보고자 하였다. 방 법 : BCG 접종 후 발생한 화농성 림프절염 환자 16명과 건강 대조군 10명의 말초 혈액에서 단핵구를 분리하고, TLR2 리간드인 Pam3CSK4로 자극한 후 유세포분석과 역전사중합효소반응을 이용하여 TLR2의 발현을 측정하였고, 자극 후 $TNF-{\alpha}$와 IL-6의 생성을 측정하여 TLR2의 발현 정도를 간접적으로 조사하였다. 결 과 : BCG 접종 후 발생한 화농성 림프절염 환자 단핵구의 TLR2 발현 정도($3.39{\pm}1.2%$)는 대조군($4.64{\pm}2.6%$)에 비하여 유의하게 감소하였고, 단핵구 자극에 의한 $TNF-{\alpha}$와 IL-6의 생성도 대조군($TNF-{\alpha}$, $1,098.5{\pm}94.3pg/mL$; IL-6, $6,696.3{\pm}544.3pg/mL$)에 비하여 환자군($TNF-{\alpha}$, $775.5{\pm}60.8pg/mL$; IL-6, $4,645.8{\pm}583.9pg/mL$)에서 유의하게 감소하였다. 그리고 자극 시간에 따른 TLR2 발현 정도와 $TNF-{\alpha}$와 IL-6의 생성 증가가 유사한 양상을 나타내었다. 결 론 : 본 연구의 결과에서 BCG 접종 후 발생한 화농성 림프절염 환자군 단핵구의 TLR2 발현 감소가 연관되어 있고, M. bovis BCG의 리간드 인식에 TLR2가 관여함을 추정할 수 있다.

• BMB Reports
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• 제53권4호
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• pp.223-228
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• 2020

Dysregulation of histone deacetylase 6 (HDAC6) can lead to the pathologic states and result in the development of various diseases including cancers and inflammatory diseases. The objective of this study was to elucidate the regulatory role of microRNA-22 (miR-22) in HDAC6-mediated expression of pro-inflammatory cytokines in lipopolysaccharide (LPS)-stimulated macrophages. LPS stimulation induced HDAC6 expression, but suppressed miR-22 expression in macrophages, suggesting possible correlation between HDAC6 and miR-22. Luciferase reporter assays revealed that 3'UTR of HDAC6 was a bona fide target site of miR-22. Transfection of miR-22 mimic significantly inhibited LPS-induced HDAC6 expression, while miR-22 inhibitor further increased LPS-induced HDAC6 expression. LPS-induced activation of NF-κB and AP-1 was inhibited by miR-22 mimic, but further increased by miR-22 inhibitor. LPS-induced expression of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 was inhibited by miR-22 mimic, but further increased by miR-22 inhibitor. Taken together, these data provide evidence that miR-22 can downregulate LPS-induced expression of pro-inflammatory cytokines via suppression of NF-κB and AP-1 axis by targeting HDAC6 in macrophages.

• 대한약침학회지
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• 제5권2호
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• pp.52-62
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• 2002

Objectives : The purpose of this study was to investigate the effects of Bee Venom on NO, $H_2O_2$ expression induced by LPS in Raw 264.7 cells as a murine marcrophage cell line and on IL-1 expression induced by LPS in D10S cells. Methods : The expression of NO was measured by MTT Assay and IL-1 by MTS Assay. The expression of $H_2O_2$ was measured as ROS level within the cell using by FACS analysis. The non-toxic concentration(from $0.1\;{\mu}g/ml\;to\;5\;{\mu}g/ml$) of Bee Venom was determined by MTT Assay. Results : 1. Bee Venom inhibited the NO expression. The effective concentration of Bee Venom was $5\;{\mu}g/ml$ after 3 hours, 1 and $5\;{\mu}g/ml$ after 1 day and 2 days. The all concentration of Bee Venom inhibited the NO expression after 6, 12 hours and 3 days. 2. Bee Venom inhibited the $H_2O_2$ expression in a dose-dependent manner compared to the control. 3. Bee Venom could not significantly inhibit the IL-1 expression.