• 제목/요약/키워드: Cell staining

검색결과 2,177건 처리시간 0.027초

해양 섬모충플랑크톤 정량과 정성분석의 현실적 접근 (Practical Approach for Quantitative and Qualitative Analyses of Marine Ciliate Plankton)

  • 김영옥;김선영;최정민;김재성
    • 한국해양학회지:바다
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    • 제26권3호
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    • pp.248-262
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    • 2021
  • 섬모충플랑크톤은 크게 두 분류군, 세포가 피갑(lorica)에 에워 쌓여 있는 유종섬모류(tintinnids)와 피갑(lorica)이 없는 소모류(oligotrichs)로 대별하고 있다. 국내에서 해양 섬모충플랑크톤의 분류학적 연구는 비교적 종 동정이 용이한 유종섬모류를 대상으로 먼저 시작되었고, 분류기준인 피갑의 가변성 때문에 분류학적 문제점은 있으나, 현재까지도 주로 피갑의 형태적 특징에 따라 종을 구별하여 생태·생리학적 연구정보로 활용하고 있다. 반면, 소모류의 경우, 세포염색을 통한 섬모열의 정밀한 특징으로 종을 구별하는 어려움으로 인해, 소모류의 분류학적 연구는 2000년 이후에 비로소 국내에 소개되었다. 최근 분석기술의 발달과 더불어 섬모충의 분류체계와 종 정보에도 많은 변화가 생겼고 이를 반영하여 국내 해양에서 출현하는 주요 섬모충플랑크톤 종류가 새롭게 정리되어 도감으로 출간되었다. 섬모충플랑크톤은 동물플랑크톤의 질 높은 먹이원이 되는 동시에 초미소(pico)와 미소(nano)플랑크톤의 포식자로서 해양 미세먹이망에서 중요한 중간 역할을 하고 있다. 이러한 부유생태계에서 섬모충플랑크톤의 역할은 새로운 주제가 아닌, 현재는 해양생태학의 보편적 지식으로 공유되고 있다. 따라서 해양 섬모충플랑크톤의 현장 모니터링 방법과 실험실 분석의 과학적 타당성을 기반으로, 획득된 자료로써 그 신뢰성을 확보함은 국내 섬모충플랑크톤 연구 발전에 크게 일조하리라 기대된다. 섬모충플랑크톤 연구 경험을 바탕으로 유종섬모류 동일종의 피갑변이, 세포염색에 의한 소모류의 정량·정성분석, 유전자 분석결과의 자료비교, 생태학적 정보비교, 섬모충플랑크톤 현장채집 방법을 제안하며, 현실적 수용가능 범위를 설정하여 섬모충플랑크톤 모니터링의 정도관리를 위한 연구자들의 관심을 유도하고자 한다.

비만마우스에서 Dextran Sulfate Sodium 유도 궤양성 대장염에 대한 커큐민의 보호효과 (Protective Effect of Curcumin on Dextran Sulfate Sodium-Induced Ulcerative Colitis in Obese Mice)

  • 박호
    • 대한임상검사과학회지
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    • 제54권1호
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    • pp.9-14
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    • 2022
  • 궤양성 대장염은 면역학적인 이상과 유전적인 요소에 의해 대장의 점막 또는 점막밑층에 염증을 유발하는 질환이다. 이전 연구에 의하면, 비만은 궤양성 대장염의 유병률을 높이고, 진행을 악화시키는 것으로 보고되었다. 따라서 본 연구에서는 커큐민이 비만에 의한 대장염의 진행을 억제하는지에 대해 조사하였다. 비만을 유도하기 위해 고지방 식이로 마우스를 사육하였고, 커큐민의 염증억제 효과를 확인하기 위해 고지방 식이와 함께 커큐민을 섭취시켰다. 궤양성 대장염을 유도하기 위해서 DSS를 경구투여 하고 대장염의 임상 증상을 관찰하였다. 대장염의 진행을 확인하기 위해 질병활성지수와 결장의 길이를 측정하였다. 커큐민에 의한 대장염 완화를 조직학적으로 평가하기 위해 결장, 간, 복부지방의 조직표본을 제작하고 H&E 및 Alcian blue-PAS 염색법으로 분석하였다. 커큐민은 질병활성지수의 점수를 감소시키고, 결장의 길이가 짧아지는 것을 억제하는 것으로 확인하였다. 또한, 비만으로 악화된 궤양성 대장염의 결장 조직에서 염증세포 침윤과 점막손상을 억제하였다. 커큐민은 간의 지방증과 복부지방의 지방세포를 유의하게 감소시키는 것으로 확인하였다. 결론적으로 커큐민은 비만에 의한 대장염의 진행을 억제함으로써 궤양성 대장염을 치료할 수 있는 치료제로서 활용될 것으로 사료된다.

증자 둥굴레 추출물의 3T3-L1 지방세포에서 분화억제 및 지질강하 효과 (Effects of steamed Polygonatum odoratum extract on inhibition of adipocyte differentiation and lowing lipid in 3T3-L1 adipocytes)

  • 강병태;최원경;박동철;김종국;박모라;김성옥;김미려
    • 대한본초학회지
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    • 제29권2호
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    • pp.15-21
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    • 2014
  • Objectives : The purpose of this study was to investigate inhibitory effects of steamed Polygonatum odoratum extract (POE) on differentiation and adipogenesis in 3T3-L1 adipocytes. Methods : Polygonatum odoratum (P. odoratum) extract was extracted with ethyl acetate. Total phenolic and flavonoid contents in POE were measured for antioxidant activity. The spectrophotometric method was used to determine the DPPH and ABTS radical scavenging activity and ferric-reducing antioxidant potential (FRAP). MTT assay was examined for cell toxicity, oil red O staining was performed for intracelluar adipogenesis in differentiated 3T3-L1 adipocytes. Western blot analysis for measurement of CCAAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor${\gamma}$ ($PPAR{\gamma}$) and AMP-activated protein kinase (AMPK) expressions were performed. Results : The results revealed that POE has antioxidant activities. Contents of total polyphenolics and flavonoids were $50.83{\pm}1.52$ GAE mg/100g dry weight of POE and $17.05{\pm}2.47$ RE mg/100g dry weight of POE, respectively. DPPH radical scavenging activity, and FRAP in 10 mg/ml concentration were $92.1{\pm}0.6%$, $244.8{\pm}9.0{\mu}M$ Fe(II) and ABTS inhibition in 5 mg/ml concentration was $84.8{\pm}4.1%$. Treatment of POE in adipocytes inhibited the differentiation and adipogenesis of 3T3-L1 adipocytes compared to those of vehicle control. Additionally, protein expressions of $C/EBP{\alpha}$ and $PPAR{\gamma}$, major transcription factor for the adipogenic genes, were significantly decreased compared to those of vehicle control (p<0.05). Futhermore, phosphorylation of AMPK was increased in 3T3-L1 adipocytes treated with POE compared to that of vehicle control (p<0.05). Conclusions : we demonstrate that steamed P. odoratum extract (POE) has potentiating antioxidant activities, inhibits differentiation and lipid accumulation and also induces energy expenditure in adipocytes, which may contribute to antiobesity property.

대시호탕의 새로운 제형이 3T3-L1에서 지방세포 증식과 분화 과정에 미치는 영향 (Effects for the New Formulation of Daesiho-tang on adipocyte development and differentiation in 3T3-L1)

  • 최혜민;김세진;문성옥;이지범;이하영;김종범;이화동
    • 대한본초학회지
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    • 제33권2호
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    • pp.69-77
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    • 2018
  • Objectives : Daesiho-tang (DSHT) has been widely used in the treatment of cerebral infarct in traditional medicine. However, there was not report on the anti-obesity-related diseases efficacy of DSHT. In this study, we investigated the effects for the new formulation of DSHT, on the adipocyte differentiation cycle in 3T3-L1 cells. Methods : 3T3-L1 cells were treated with DSHT (50, 100, $200{\mu}g/m{\ell}$) during differentiation for 6 days. Also, the inhibitory effect of DSHT against 3T3-L1 adipogenesis was evaluated in various stage of adipogenesis such as early (0-2day), intermediate (2-4day), and terminal stage (4-6day). The accumulation of lipid droplets was determined by Oil Red O staining. and, the expressions of genes related to adipogenesis were measured by RT-PCR and Western blot analyses. Results : DSHT showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affect cell toxicity as assessed by measuring fat accumulation and adipogenesis. In addition, DSHT significantly reduced the expression levels of several adipocyte marker genes including proliferator activated $receptor-{\gamma}$ ($PPAR-{\gamma}$) and CCAAT/ enhancer-binding $protein-{\alpha}$ ($C/EBP-{\alpha}$). Also, the anti-adipogenic effect of DSHT was strongly limited in the intermediate (2-4 day), terminal stage (4-6 day) of 3T3-L1 adipogenesis. In addition, the DSHT treatment down- regulated mRNA expression levels of $PPAR-{\gamma}$,, $C/EBP-{\alpha}$ in mature 3T3-L1 adipocytes. Conclusions : These results suggest that, the ability of DSHT has inhibited overall adipogenesis and lipid accumulation in the 3T3-L1 cells. The new formulation of DSHT may be a promising medicine for the treatment of obesity and related metabolic disorders.

3T3-L1 전구지방세포에서 개구리자리(Ranunculus sceleratus) 추출물의 AMPK 신호전달을 통한 지방생성 억제 효과 (Extract of Ranunculus sceleratus Reduced Adipogenesis by Inhibiting AMPK Pathway in 3T3-L1 Preadipocytes)

  • 김예지;조성필;이희주;홍금란;김경현;류시윤;정주영
    • 한방비만학회지
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    • 제22권1호
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    • pp.30-37
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    • 2022
  • Objectives: Adipogenesis is the process by which pre-adipocytes are differentiated into adipocytes. It also plays an important role in adipocyte formation and lipid accumulation. Ranunculus sceleratus (R. sceleratus) extracts are used for the treatment of various diseases such as hepatitis, jaundice, and tuberous lymphadenitis in oriental medicine. However, its effect on adipogenesis has not yet been studied. In this study, we investigated the effects of R. sceleratus on adipogenesis in 3T3-L1 cells. Methods: Cells were treated with 50, 100, and 200 ㎍/ml of R. sceleratus and cell viability was evaluated. To differentiate the 3T3-L1 preadipocytes, a 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI) solution were used. The accumulation of lipid droplets was determined by Oil Red O staining. The expression levels of adipogenesis-related proteins were also determined. Results: MDI solution differentiated the preadipocytes into adipocytes and accumulation of lipids was observed in the differentiated 3T3-L1 cells. Interestingly, the amount of lipid droplets was reduced after R. sceleratus treatment. In addition, the expression levels of key adipogenic transcription factors, such as CCAAT/enhancer-binding proteins-𝛼 (C/EBP-𝛼) and peroxisome proliferator-activated receptors-𝛾 (PPAR-𝛾) were also reduced after R. sceleratus treatment. Furthermore, R. sceleratus increased AMP-activated kinase (AMPK) phosphorylation and decreased sterol regulatory element-binding protein-1 expression. Conclusions: Our results showed that R. sceleratus reduced preadipocyte differentiation by inhibiting C/EBP-𝛼 and PPAR-𝛾 levels via the AMPK pathway. Therefore, we suggest that R. sceleratus may be potentially used as an anti-adipogenic agent.

노각나무 가지 추출물이 Porphyromonas gingivalis의 바이오필름 형성에 미치는 영향 (Effect of Stewartia koreana Nakai Branch Extracts on the Biofilm Formation of Porphyromonas gingivalis)

  • 박민정;김혜수;김한비;박준호;유찬열;조수정
    • 생명과학회지
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    • 제32권1호
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    • pp.56-62
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    • 2022
  • 본 연구에서는 천연물유래 구강건강 개선소재로써 노각나무 가지의 이용 가능성을 검토하기 위해 노각나무 가지 추출물이 치주염 원인균인 P. gingivalis KCTC5352의 병원성인자(virulence factor)에 미치는 영향을 조사하였다. 추출물은 P. gingivalis에 대해 정균작용을 나타내었고 P. gingivalis에 대한 추출물의 MIC는 0.6 mg/ml였다. 추출물 처리구(0.2-2.0 mg/ml)에서 P. gingivalis의 병원성 인자인 바이오필름은 추출물의 농도가 증가할수록 농도 의존적으로 억제되는 경향을 나타내었고, 주사전자현미경으로 추출물 처리구의 P. gingivalis를 관찰한 결과에 의하면 추출물 무처리구에서는 세포들의 응집(autoaggregation)과 바이오필름이 관찰되었지만 추출물 처리구에서는 세포들의 응집과 바이오필름이 관찰되지 않았다. qRT-PCR을 이용하여 P. gingivalis의 바이오필름 형성에 필수적인 섬모(fimbriae) 유전자에 대한 mRNA 발현을 조사한 결과에서도 섬모관련 유전자인 fimA와 fimB에 대한 mRNA 발현은 추출물의 농도가 높아질수록 농도의존적으로 감소되는 것을 확인할 수 있었다. 이상의 결과를 종합하면 노각나무 가지 추출물은 치주염 원인균인 P. gingivalis에 대해 정균작용을 나타낼 뿐만 아니라 P. gingivalis의 병원성 인자인 바이오필름의 형성을 효과적으로 억제할 수 있기 때문에 천연물유래 구강건강 개선소재로써 이용 가능성이 높을 것으로 판단된다.

Ginsenoside Rb2 suppresses cellular senescence of human dermal fibroblasts by inducing autophagy

  • Kyeong Eun Yang;Soo-Bin Nam;Minsu Jang;Junsoo Park;Ga-Eun Lee;Yong-Yeon Cho;Byeong-Churl Jang;Cheol-Jung Lee;Jong-Soon Choi
    • Journal of Ginseng Research
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    • 제47권2호
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    • pp.337-346
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    • 2023
  • Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

Noni Fruit Juice의 3T3-L1 지방전구세포 분화 억제 및 인터루킨-1β 유도 Inducible Nitric Oxide Synthase 염증유전자 발현 감소 효과 (Suppressive Effects on Lipid Accumulation and Expression of Interleukin-1β-Mediated Inducible Nitric Oxide Synthase in 3T3-L1 Preadipocytes by a Standardized Commercial Noni Fruit Juice)

  • 장병철
    • 한방비만학회지
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    • 제23권1호
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    • pp.1-9
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    • 2023
  • Objectives: Noni fruit juice (NFJ) is liquor extracted from Morinda citrifolia (noni) fruit and has been used as an herbal remedy in many countries. However, the NFJ's anti-adipogenic and anti-inflammatory effects on adipocytes are poorly understood. The purpose of this study was to explore the commercially standardized NFJ effects on lipid accumulation throughout 3T3-L1 preadipocytes differentiation and interleukin-1β (IL-1β)-mediated inducible nitric oxide synthase (iNOS) expression in 3T3-L1 preadipocytes. Methods: Cellular lipid accumulation and triglyceride (TG) content in differentiating 3T3-L1 preadipocytes were assessed subsequently via the Oil Red O staining and AdipoRed assay. MTS assay was used to examine NFJ cytotoxicity in (differentiating) 3T3-L1 preadipocytes. Immunoblotting and reverse transcriptase polymerase chain reaction analysis were used to measure the expression levels of target protein and mRNA in (differentiating) 3T3-L1 preadipocytes, respectively. Results: NFJ treatment at 150 μL/mL led to a substantial reduction of fat accumulation and TG content during 3T3-L1 adipogenesis with no discernable impact on the cell viability. Of note, while NFJ treatment (150 μL/mL) largely inhibited the CCAAT/enhancer-binding protein-α (C/EBP-α) and peroxisome proliferator-activated receptor-β (PPAR-β) protein expressions, it did not influence PPAR-γ in differentiating 3T3-L1 preadipocytes. Of interest, treatment with IL-1β at 20 ng/mL for 4 hours elicited in firm induction of iNOS mRNA expression in 3T3-L1 preadipocytes. However, NFJ treatment at 100 or 200 μL/mL greatly attenuated the IL-1β-induced iNOS mRNA expression in 3T3-L1 preadipocytes. Conclusions: NFJ has anti-adipogenic and anti-inflammatory effects on (differentiating) 3T3-L1 preadipocytes which are in part intervened via control of the expression of C/EBP-α, PPAR-β, and iNOS.

당귀작약산의 창상 회복에 대한 효과 (The Effect of Dangguijakyak-san on Wound Healing)

  • 이윤진;우창훈;김영준;김현지;안희덕
    • 한방재활의학과학회지
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    • 제33권3호
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    • pp.47-65
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    • 2023
  • Objectives We evaluated the wound healing effects of Dangguijakyak-san (DJ) using C57BL/6 mice that were generated open wound. Methods The study was conducted with seven C57BL/6 mice assigned to each group, divided into the normal group, control group, vitamin E group, DJ low-dose group, DJ high-dose group. We measured total polyphenol, flavonoid contents, the size of the wound, liver function, pro-inflammatory cytokine activity in serum, inflammation-related proteins, adhesion molecules and chemokine proteins, collagen-related proteins in skin tissue and histopathological changes by H&E and Masson's staining. Results DJ treatment significantly reduced the area of the wound compared to the control group. Also, inflammatory cytokines were reduced and the expression of anti-inflammatory-related factors (interleukin-4 [IL-4] and IL-10) was significantly increased in the DJ treatment group. We identified that DJ treatment inhibits both pathways of inflammation, the mitogen-activated protein kinases and nuclear factor-κB pathway. Moreover, the protein expressions of Sirt1 (sirtuin 1), MCP-1 (monocyte chemoattractant protein 1), ICAM-1 (intercellular adhesion molecule 1), and VCAM-1 (vascular cell adhesion molecule 1) were decreased by DJ administration. Also, the expression of α-smooth muscle actin and collagen type I alpha 1, collagen-related proteins, that help skin recovery was significantly increased in the DJ treatment group. Histopathologically, a relatively thin epithelial layer could be observed in the DJ administration group, as well as an increase in fibroblasts and collagen fibers. Conclusions These data suggest that DJ treatment is effective in wound healing, suppressing inflammatory proteins, increasing skin repair factors and improving histopathological changes caused by wounds.

노각나무 추출물이 Pseudomonas aeruginosa의 바이오필름 형성에 미치는 영향 (Inhibitory Effects of Stewartia koreana Extracts on Pseudomonas aeruginosa Biofilm Formation)

  • 이상균;김혜수;조수정
    • 생명과학회지
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    • 제33권11호
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    • pp.936-943
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    • 2023
  • 본 연구에서는 천연물 유래 항균소재로써 노각나무의 이용 가능성을 알아보기 위해 노각나무 가지, 줄기, 잎을 에탄올에 추출한 다음 P. aeruginosa에 대한 추출물의 항균활성을 조사하였다. 노각나무 부위별 추출물(1 mg/disc) 중 잎과 줄기 추출물이 P. aeruginosa에 대해 항균활성을 나타내었으며, 잎 추출물의 항균활성이 가장 우수하였다. P. aeruginosa에 대한 노각나무 잎 추출물의 MIC는 0.8 mg/ml였고 정균작용을 나타내었다. 노각나무 잎 추출물이 0.2-2.0 mg/ml 농도로 처리된 배양액에서 P. aeruginosa의 바이오필름 형성은 추출물의 농도가 증가할수록 농도의존적으로 억제되는 경향을 보였다. 또한 주사전자현미경으로 노각나무 잎 추출물(0.8 mg/ml)이 P. aeruginosa의 바이오필름 형성에 미치는 영향을 관찰한 결과에 의하면 추출물 대신 DMSO를 처리한 대조구에서는 세포 주변에서 EPS와 바이오필름이 관찰되었지만 추출물을 처리한 처리구의 세포 주변에서는 EPS와 바이오필름이 관찰되지 않았다. qRT-PCR을 이용하여 P. aeruginosa의 QS 관련 유전자인 lasI와 rh1I mRNA 발현 변화를 qRT-PCR로 확인한 결과, 노각나무 잎 추출물이 0.2-2.0 mg/ml의 농도로 처리된 배양액에서 lasI와 rh1I mRNA 발현은 추출물의 농도가 높아질수록 농도의 존적으로 억제되는 것을 확인할 수 있었다. 이상의 결과를 종합하면 노각나무 잎 추출물은 P. aeruginosa에 대한 항균활성과 바이오필름 형성 억제능이 우수하기 때문에 천연물 유래 항균소재로써 이용 가능성이 높을 것으로 판단된다.