Antioxidative activity of Chicoric acid through Nrf2/MAPK mediated HO-1 induction in RAW 264.7 cells

박충무 사단법인 인문사회과학기술융합학회 2017 예술인문사회융합멀티미디어논문지 Vol.7 No.4

The antioxidativepotential of chicoric acid was investigated through heme oxygenase (HO)-1, one of phase II enzymes and exhibits antioxidative activity, induction in RAW 264.7 cells. There were also measured intracellular radical scavenging activity and glutathione (GSH) concentration in chicoric acid treated RAW 264.7 cells. Chicoric acid treatment potently induced HO-1 expression, scavenged reactive oxygen species (ROS) and increased GSH concentration. Transcription factor of phase II enzymes, nuclear factor-erythroid 2 p45-related factor 2 (Nrf2), was investigated and identified in accordance with HO-1 expression. In addition, chicoric acid treatment phosphorylated PI3K/Akt with slight activation of the extrecellularregulated kinase (ERK). Chicoric acid-induced antioxidative potential protected RAW 264.7 cells against oxidative stress-induced cell death. In conclusion, chicoric acid potently accelerated Nrf2-mediated antioxidative potential through the modulation of PI3K/Akt pathway, which would contribute to their promising strategy for a novelanti-oxidative agent.

성인들의 건강관리와 구강건강관리행태에 따른 주관적 건강과 구강건강, EQ-5D수준

박충무,윤현서 대한통합의학회 2023 대한통합의학회지 Vol.11 No.1

Purpose : This study examined the relationship between oral health management behavior and subjective health perception by using a tool that measures health-related quality of life named EQ-5D. The findings are expected to provide fundamental data for developing diverse programs, including health- and oral-health-related educational media, to ultimately enhance people’s health-related quality of life. Methods : Adults over the age of 20 were extracted from the raw materials of the 8th national health and nutrition examination survey conducted in 2019. Among them, 4034 were finally chosen as the study subjects after checking the response distribution of relevant questions and excluding inadequate samples. Variables included general characteristics, health management behavior, oral health management behavior, subjective health perception, and EQ-5D level. Cross-tabulation test, independent t-test, one-way analysis of variance, and linear regression analysis were conducted using SPSS 25.0. Results : EQ-5D, which measures the level of health-related quality of life, was high when the subjects had a medical check-up experience, no experience of unsatisfactory medical service, and did not get an influenza vaccination (p<.001). The quality of life was higher when the subjects received dental inspections, did not require treatment, and used oral care goods (p<.005). Health-related quality of life was also higher when the subjective health level and subjective oral health level were high. Conclusion : Results indicate that subjective health, oral health perception level, and quality of life together were high when people had a regular check-up, did not neglect treatment, and used oral care goods. Considering the findings of this study, check-up programs that are customized for each age group are necessary. Health-related quality of life could be enhanced through the provision of diverse medical services and active efforts to prevent any medical blind spot.

사람 치은섬유모세포에서 NF-κB와 JNK 활성 억제를 통한 돌김 에탄올 추출물의 항염증 효과

박충무,윤현서 한국융합학회 2018 한국융합학회논문지 Vol.9 No.12

Human gingival fibroblast (HGF) is the main cell type existed in periodontium and produces a variety of inflammatory mediators by external stimuli. In this study, the anti-inflammatory activity of Porphyra yezoensis ethanol extract (PYEE) on LPS-PG lipopolysaccharide from Porphyromonas gingivalis activated HGF-1 cell. Up-regulated iNOS and COX-2 expressions by LPS-PG were significantly attenuated by PYEE treatment in a dose-dependent manner. In addition, activated nuclear factor (NF)-κB was also dose-dependently inhibited by PYEE treatment. Among upstream signaling molecules, PYEE treatment inhibited phosphorylation of c-Jun NH2-terminal kinase (JNK) but did not give any effect on other molecules. On the other hand, one of phase II enzymes, NAD(P)H:quinone dehydrogenase (NQO)-1, was analyzed due to its anti-inflammatory activity, which was upregulated by PYEE treatment. Consequently, PYEE could be candidates for the prevention and treatment of periodontal diseases. 사람치은섬유모세포(human gingival fibroblast, HGF)는 치은조직에 존재하는 주요한 세포의 형태 중 하나로 외부 자극에 반응하여 다양한 염증대사물질을 생산한다. 본 연구에서는 돌김에탄올추출물(PYEE)이 Porphyromonas gingivalis로부터 분리한 lipopolysaccharide로 염증이 유도된 HGF-1 cell에서 항염 효과를 보이는지 분석하고자 하였다. LPS-PG에 의해 과발현된 iNOS와 COX-2는 PYEE의 처리에 의해 농도 의존적으로 발현이 감소되었고, nuclear factor (NF)-κB 또한 동일한 양상으로 활성이 억제되었다. 신호전달물질 중 c-Jun NH2-terminal kinase (JNK)의 인산화만이 PYEE에 의해 억제되었다. 그리고 항염 작용에 관여하는 것으로 알려진 2상 효소 중 하나인 NAD(P)H:quinone dehydrogenase (NQO)-1도 분석하였고, 이 효소는 PYEE의 처리에 의해 강하게 발현이 유도가 되었다. 결론적으로 돌김에탄올추출물은 치주질환 에방과 치료를 위한 후보물질로 활용 가능할 것으로 사료된다.

Luteolin Inhibits Inflammatory Responses by Downregulating the JNK, NF-κB, and AP-1 Pathways in TNF-α Activated HepG2 Cells

박충무,진경숙,조정원,이용우,허경혜,차연수,송영선 한국식품과학회 2012 Food Science and Biotechnology Vol.21 No.1

The inhibitory mechanism of luteolin on tumor necrosis factor (TNF)-α-induced inflammation was investigated in HepG2 cells. Luteolin significantly suppressed TNF-α-stimulated inducible nitric oxide synthase (iNOS) expression in a dose-dependent manner without cytotoxicity. Phosphorylation and nuclear translocation of both transcription factors, nuclear factor (NF)-κB and activator protein (AP)-1, were also inhibited by luteolin treatment. Additionally,luteolin suppressed TNF-α-induced c-Jun N-terminal kinase (JNK) phosphorylation, which is crucially related to regulating inflammation. SP600125, a JNK selective inhibitor, abolished the TNF-α triggered inflammatory signaling cascade. These results suggest that luteolin attenuates inflammatory responses by blocking NF-κB and AP-1 activation through suppressed JNK phosphorylation in TNF-α-stimulated HepG2 cells.

Antibacterial Activity and Inhibitory Effect of Biofilm Formation by Actinidia polygama against S. mutans and P. gingivalis

박충무,윤현서 대한의생명과학회 2023 Biomedical Science Letters Vol.29 No.2

In oriental medicine, the fruit of Actinidia polygama has long been used to alleviate the symptoms of gout, arthritis, and inflammation. In this study, it was to designed to analyze the antibacterial activity of A. polygama ethanol extract (APEE) against Streptococcus mutans, one of the major strains for dental caries, and Porphyromonas gingivalis, one of the critical strains for periodontal disease. The antibacterial activity of APEE was analyzed by disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. In addition, it was also analyzed the inhibitory effect of APEE on bacterial growth and biofilm formation against both oral pathogens. APEE exhibited its antibacterial effect through the inhibited bacterial diffusion as well as low concentration of MIC and MBC. In addition, APEE significantly inhibited not only bacterial growth but also biofilm formation in a dose-dependent manner. Consequently, APEE showed potent antibacterial activity against both S. mutans and P. gingivalis, which indicates that APEE might be used as a potential antibacterial material for the improvement of oral healthcare.

사람 치은섬유모세포에서 잎꼬시래기 에탄올 추출물의 항염증 및 항산화 효과

박충무,윤현서 대한통합의학회 2019 대한통합의학회지 Vol.7 No.4

Purpose : Human gingival fibroblast cell is one of the the main cell types in periodontal tissue, which they can show anti-inflammatory activity through the production of numerous lines of inflammatory mediators. Methods : This study aimed to investigate antiinflammatory and antioxidative activities of Gracilaria textorii ethanol extract (GTEE) in Porphyromonas gingivalis derived lipopolysaccharide (LPS-PG) stimulated in HGF-1 cells. Result : Elevated inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 activities by LPS-PG stimulation were significantly attenuated by GTEE reatment in a dose-dependent manner. Intracellular NOS activity was also coincided with iNOS result. Phosphorylated status of inflammatory transcription factors, nuclear factor (NF)-κB and activator protein-1 (AP-1), were analyzed and significantly ameliorated by GTEE treatment. In addition, one of phase II enzyme, NAD(P)H quinone dehydrogenase (NQO)-1, and its transcription factor, Nuclear factor erythroid 2-related factor 2 (Nrf2), were significantly induced by GTEE treatment. Conclusion : In conclusion, GTEE mitigated LPS-PG-induced inflammatory responses by attenuated NF-κB and AP-1 activations as well as by accelerating NQO1 and Nrf2 expression in HGF-1 cells. 치주조직에 존재하는 주요한 세포의 한 형태인 치은섬유모세포는 LPS로부터 유래하는 염증성 자극에 반응하여 여러 염증 매개 물질을 분비한다. 본 연구에서는 치주염을 유발하는 원인균 중 하나인 P. gingivalis로부터 분리한 LPS를 이용하여 사람 치은섬유모세포인 HGF-1 세포에 염증을 유도한 후 GTEE의 항염증능과 항산화능을 분석하였다. 그 결과 LPS-PG에 의해 과발현된 iNOS와 COX-2는 GTEE의 처리에 의해 농도 의존적으로 발현이 억제되었고, 전사인자인 NF-κB와 AP-1의 인산화 또한 동일한 경향으로 발현이 억제되었다. 그리고 발현의 유도가 항염증 효과를 나타내는 것으로 알려진 2상 효소 중 하나인 NQO1과 이의 전사인자인 Nrf-2를 분석한 결과 GTEE의 처리에 의해 효소의 활성이 높아지는 것을 확인할 수 있었다. 결론적으로 GTEE는 NF-κB와 AP-1의 인산화를 억제하고 NQO1 활성을 유도함으로써 HGF-1 세포에서 LPS-PG에 의해 유도된 염증을 억제하는 것으로 사료되고, GTEE는 치주염 억제에 효과적인 항염증 물질의 가능성이 있는 후보물질이 될 수 있을 것으로 생각한다.

일부지역 성인 환자의 주관적 건강인식과 구강건강관리행태가 치과공포감에 미치는 영향

박충무,윤현서 한국융합학회 2018 한국융합학회논문지 Vol.9 No.12

The purpose of this study was to examine the oral health care and self-rated health status of adults visiting dental clinics in the region of Busan and influential factors for their dental fear from January to May, 2017, in an effort to obtain some information on how to relieve fear. As for dental fear, adults who were in their 30s, who were homemakers and whose monthly income ranged from 4 to 4.99 million won. In regard to fear according to oral health care, dental fear caused by the treatment avoidance factor was stronger among the adults who didn't receive dental checkups and who brushed their teeth in the wrong way. Dental fear that was attributed to the physiological reaction factor and the stimuli-inducing factor was severer among the adults who didn't receive dental checkups and who had no scaling experience. Overall fear was severer among the adults who didn't receive dental checkups and who changed their toothbrushes every four or more months. The factors that affected dental fear were self-rated health status, self-rated oral health status, gender, age, whether to receive dental checkups on a regular basis or not, and oral health education experience. Therefore in order to alleviate dental fear, self-rated health status should be improved, and the kind of system that encourages regular dental checkups and provides oral health education should be prepared. 본 연구는 2017년 1월부터 5월까지 일부지역치과에 내원하는 성인들을 대상으로 구강건강관리행태와 주관적 건강인식수준을 파악하고 치과공포감에 미치는 영향요인을 파악하여 치과공포감을 낮추기 위한 기초자료를 마련하고자 한다. 치과공포에서 회피요인은 여자, 40대, 고졸이하, 주부, 300-399만원, 생리적 반응요인은 여자, 40대, 주부, 200-299점, 자극유발유인 여자, 30대, 주부, 전체공포수준은 30대, 주부, 400-499만원에서 높았다. 구강건강관리행태에 따른 공포감에서 치료회피요인은 구강검진을 하지 않고, 칫솔질방법이 틀린 경우, 생리적 반응요인과 자극반응요인은 검진을 하지 않고, 스케일링 경험이 없는 경우, 전체공포감은 검진을 하지 않고, 칫솔교환시기가 4개월 이상인 경우에서 치과공포감이 높았다. 치과공포에 영향을 미치는 주관적 건강인식수준, 주관적 구강건강인식수준, 성별, 나이, 정기검진유무, 구강보건교육 경험유무가 영향을 미쳤다. 따라서 치과공포감을 줄이기 위해서는 주관적 인식을 개선하고 정기검진과 구강보건교육을 위한 시스템이 마련되어야 할 것이다.

RAW 264.7 세포에 대한 NF-κB와 MAPK 활성 억제를 통한 개다래 열매 에탄올 추출물의 항염증 효과

박충무,윤현서 대한통합의학회 2023 대한통합의학회지 Vol.11 No.2

Purpose : The fruit of Actinidia polygama has been used in oriental medicine for the treatment of gout, rheumatoid arthritis, and inflammation. Though A. polygama exhibited anti-inflammatory activity in RAW 264.7 cells and carrageenan-induced rat paw edema, the exact mechanism for anti-inflammation was not evaluated yet. In this study, the anti-inflammatory mechanisms of A. polygama ethanol extract (APEE) in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Methods : WST-1 assay was applied to analyze the cytotoxic effect of APEE in RAW 264.7 cells. The productions of nitric oxide (NO) and prostaglandin (PG) E2 were analyzed by the Griess reaction and enzyme immunoassay (EIA) assay, respectively. In addition, protein expressions for inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 were measured by Western blot analysis. The activated status of an inflammatory transcription factor, NF-κB, and its upstream signaling molecules, mitogen-activated protein kinases (MAPKs), was also evaluated by Western blot analysis. Results : As a result, APEE treatment did not exhibit any cytotoxicity until the concentration of 200 ㎍/㎖. APEE treatment significantly inhibited NO and PGE2 productions as well as their enzymes, iNOS and COX-2 in a dose-dependent manner. The inflammatory transcription factor, NF-κB, was also attenuated by APEE treatment. In addition, the phosphorylated status of MAPKs such as extracellular regulated kinase (ERK), c-jun NH2 kinase (JNK), and p38, were significantly diminished by APEE treatment in LPS stimulated RAW 264.7 cells. Conclusion : Consequently, APEE treatment significantly attenuated the production of inflammatory mediators and their enzyme expressions in LPS-stimulated RAW 264.7 cells. The inflammatory transcription factor, NF-κB, and upstream signaling molecules, MAPKs, were also significantly attenuated by APEE treatment in LPS-activated RAW 264.7 cells. These results indicate that APEE might be a candidate to be utilized as a promising candidate for the treatment of inflammatory disorders.

당뇨병 진단을 받은 65세 이상 노인의 구강관리행태와 EQ-5D index

박충무,윤현서 국제차세대융합기술학회 2022 차세대융합기술학회논문지 Vol.6 No.11

65세 이상 당뇨병 진단을 받은 노인들의 구강관리행태를 파악하고 삶의 질과 관련성을 확인하여 맞춤형 구강건강관리 프로그램 개발의 기초자료로 활용하고자 한다. 제 8기 국민건강영양조사 자료(2019년)를 활용하여 65 세 이상으로 당뇨병진단을 받은 대상자 356명을 대상으로 하였다. 구강검진율 22.2%, 치아손상 경험률 12.6%, 저 작불편호소률 37.6%, 치과 미치료 경험률 25.0%, 구강관리용품 사용 경험률 30.6%였고, 저작불편호소 경험이 있고 (p<.001) 치과 미치료 경험이 있는 경우 주관적 건강수준이 낮았다(p=.014). 저작불편호소 경험이 있고(p<.001) 치 과 미치료 경험이 있는 경우 주관적 구강건강수준이 낮았다(p=.001). 구강건강관리행태에 따른 EQ5Dindex는 구강 검진을 하는 경우(p<.001)와 저작불편호소 경험이 없는 경우(p<.001)에 높게 나타났다. EQ5Dindex는 주관적 건강 수준 인식이 높고, 구강검진 경험이 있는 경우 양의상관관계를 보였고, 주관적 건강인식은 주관적 구강건강인식수 준이 높은 경우 양의 상관관계를 보였으며, 구강검진 여부에서는 미치료 경험이 없는 경우 양의 상관관계를 보였 다. 따라서 치주질환에 취약한 당뇨병 환자들을 대상으로 하는 맞춤형 구강보건교육과 관리법을 제공하여 삶의질 향상을 도모할 수 있을 것으로 생각된다. This study is tried to provide the fundamental data for the development of customized oral health management program to 65 years of age or older with diagnosed as diabetes through identifying oral care behavior and the relevance to quality of life. Three hundred fifty six subjects, diagnosed as diabetes, were analyzed, which was substracted from 8th National Health and Nutrition Survey data (2019). Results are as follows; 22.2% of the oral medical checkup rate, 12.6% of tooth damage experience rate, 37.6% of the discomfort of chewing rate, 25.0% of dental untreated rate, 30.6% of oral care product use rate. Cases of low subjective health level are in discomfort of chewing (p<.001) and dental untreated experience (p=.014). EQ5D index according to oral care behavior was high when oral examination (p<.001) is performed (p<.001) and when there is no experience of chewing discomfort. In addition, EQ5D index exhibited a positive correlations in case of high subjective health awareness level and dental untreated experience. Consequently, it might be considered that the way to promote the quality of life through the supply of customized oral health education and management for diabetic patients who are vulnerable to periodontal disease.

청견 잎 에탄올 추출물의 NF-κB와 MAPK 조절을 통한 항염증 효과

박충무,윤현서 대한통합의학회 2023 대한통합의학회지 Vol.11 No.3

Purpose : Though other Citrus spp. have reported their anti-inflammatory and antioxidative activities in previous studies, the biological activity of Kiyomi (Citrus unshiu x C. sinensis) has not been reported yet. Therefore, this study attempted to analyze the anti-inflammatory mechanisms of Kiyomi leaf ethanol extract (KLEE) in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Methods : The cytotoxic effect of KLEE in RAW 264.7 cells was determined by WST-1 assay. Bacterial endotoxin, the concentration of nitric oxide (NO) was analyzed by the Griess reaction. In addition, Western blot analysis was applied to measure the protein expression level of inducible NO synthase (iNOS). The phosphorylated status of the critical inflammatory transcription factor, nuclear factor (NF)-κB, and its upstream signaling molecules, phosphoinositide 3-kinase (PI3K)/Akt as well as mitogen-activated protein kinases (MAPKs), were also measured by Western blot analysis. Results : KLEE was not cytotoxic up to a concentration of 200 ㎍/㎖, and protein expression levels of iNOS and cyclooxygenase (COX)-2, enzymes that counteract NO and prostaglandin (PG) E2 production, were inhibited by KLEE treatment. The phosphorylated status of PI3K/Akt as well as MAPKs including extracellular regulated kinase (ERK), c-jun NH2kinase (JNK), and p38, were significantly attenuated by KLEE treatment in LPS stimulated RAW 264.7 cells. Moreover, one of phase II enzymes, heme oxygenase (HO)-1 which has known for its anti-inflammatory capacity, was strongly induced by KLEE treatment. Conclusion : Consequently, KLEE treatment significantly attenuated the production of NO as well as the expression levels of iNOS and COX-2 in LPS-stimulated RAW 264.7 cells. The inflammatory transcription factor, NF-κB, as well as its upstream signaling molecules, PI3K/Akt and MAPKs, were also diminished by KLEE treatment with statistical significance in LPS-stimulated RAW 264.7 cells. These results suggest that KLEE might be a promising candidate for the attenuation of inflammatory disorders.