• Journal of Microbiology and Biotechnology
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• 제34권6호
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• pp.1340-1347
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• 2024

Ehretia asperula is a medicinal plant of the Ehretiaceae family used to treat inflammatory disorders, but the underlying mechanisms are not fully elucidated. The anti-inflammatory potential was determined based on enzyme cyclooxygenase-2 (COX-2) inhibition, which showed that the 95% ethanol extract (95ECH) was most effective with a half-maximal inhibitory concentration (IC₅₀) value of 34.09 ㎍/mL. The effects of 95ECH on phagocytosis, NO production, gene, and protein expression of the cyclooxygenase 2/prostaglandin E2 (COX-2/PGE2) and inducible nitric oxide synthase/ nitric oxide (iNOS/NO) pathways in lipopolysaccharide (LPS)-induced RAW264.7 cells were examined using the neutral red uptake and Griess assays, reverse-transcriptase polymerase chain reactions (RT-PCR), and enzyme-linked immunosorbent assays (ELISA). The results showed that 95ECH suppressed phagocytosis and the NO production in activated macrophage cells (p < 0.01). Conversely, 95ECH regulated the expression levels of mRNAs for cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) as well as the corresponding proteins. In addition, PGE2 production was inhibited in a dose-dependent manner by 95ECH, and the expression of iNOS and COX-2 mRNAs was decreased in activated macrophage cells, as expected. Therefore, 95ECH from E. asperula leaves contains potentially valuable compounds for use in inflammation management.

• 식물병연구
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• 제12권2호
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• pp.91-98
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• 2006

Acinetobacter sp. KTB3의 배양여과액으로 제조한 분말 제제 KNF2022의 바이러스 감염억제 효과를 PMMoV 와 N. glutinosa 또는 N. tabacum cv. Xanthi nc를 이용하여 검정한 결과, 증류수 1/100 배양액의 실질 농도 10,000 ppm) 희석농도에서 $94.3{\sim}95.6%$의 높은 감염억제 효과를 나타냈다. 이 1/100 희석농도를 이용하여 PepMoV-C amaranticolor에서 검정한 억제효과는 97.1%, CMV-C amaranticolor 에서는 92.5%의 감염억제를 나타냈다. KNF2022 희석액을 N. glutinosa의 반엽에 도말하고 일정 시간 경과 후 PMMoV를 접종하여 조사한 억제효과의 지속성은 처리 2 일 후까지 약 80% 선에서 억제효과를 나타냈으나, 처리 4 일 후부터는 50% 수준으로 저하되었다. 고추에 KNF2022를 처리한 후 3 종의 바이러스를 접종하고 발현되는 병징을 조사한 결과, 각 바이러스를 단독으로 접종하였을 경우는 바이러스의 종류에 관계없이 접종 후 10 일까지 병징발현이 지연되는 효과를 보였다. 특히 PepMoV를 접종한 경우는 접종 후 30 일까지 병징발현이 억제되어 제제의 처리효과가 뚜렷하게 나타났다. 한편 이들 3 종 바이러스의 복합감염에 대한 KNF2022의 효과는 모든 바이러스 조합에서 단독감염 보다 강하게 발현되어 병징억제의 효과는 뚜렷하게 나타나지 않았다. 제제를 처리한 고추에서 증식된 바이러스의 농도를 PCR 및 ELISA로 검출한 결과, PepMoV를 단독으로 접종한 경우, 접종 후 30 일까지 cDNA가 검출되지 않았다. RT-PCR 검정에서 억제효과가 인정된 PepMoV와 그 복합감염 의 조합에 대해서 ELISA 검정을 실시한 결과, PCR 검정에서 얻어진 결과와 유사한 패턴을 보여 PepMoV에 대한 감염억제효과가 뚜렷하게 인정되었다. KNF2022를 처리하여 전자현미경으로 관찰한 PMMoV와 PepMoV는 처리 5 분 후에 각각 200-250 nm 및 400-600 nm의 길이로 절단된 입자가 많이 관찰되었고, 처리 30 분 후에는 대부분 100-150nm와 300-500 nm의 절편입자로 관찰되었다. 이와 같이 절단된 입자가 증가할수록 N. tabacum cv. Xanthi nc 와 C. amaranticolor에서 나타난 병반수는 급격하게 감소되었다. 한편 KNF2022를 처리한 후 전자현미경을 관찰한 CMV의 바이러스 입자는 처리시간에 비례하여 파괴된 입자의 수가 증가하였으며, 파괴된 바이러스입자가 증가할수록 C. amaranticolor에서의 병반수도 감소되었다.

• 분석과학
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• 제18권2호
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• pp.104-111
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• 2005

70% 에탄올로 추출한 후 농축한 유근피 추출물을 여러 용매로 분획한 부분들의 MMP-9 활성 저해능 실험에서 ethyl acetate 분획분이 MMP-9 활성 저해능이 있는 것으로 나타났다. 이 분획분에서 MMP-9의 활성에 저해능을 나타내는 물질을 분리 정제하여 분석하였고 이 물질은 catechin 계열로 판명되었다. Zymography법에 의해 MMP-9의 활성 억제 효과를 관찰하였고 정제된 화합물은 MMP-9을 5 mM 농도에서 48%가 억제하였고 10 mM 에서는 43%를 억제하는 것으로 나타났다. 정상 세포주인 Chang 세포주와 인체 간암세포인 SK-Hep-1 세포주를 동일한 조건으로 배양한 후에 형태학적 변화를 관찰한 결과, 정상 세포주의 핵과 그 주변에서는 변화가 거의 관찰되지 않았으나 SK-Hep-1 세포주는 핵의 주위에 검은 반점이 관찰되었다. 세포의 증식과 살아있는 세포를 측정하기 위하여 MMP-9 억제물질인 정제된 추출물을 투여하고 MTT 검색법에 의해 ELISA reader로의 측정에서 정상세포주인 Chang 세포주에서는 1 mM에서 세포 활성이 오히려 증가하는 경향($130.77{\pm}13.71$)을 보인 반면, SK-Hep-1 세포주에서는 세포 증식이 상당히 억제된 것으로 나타났다.

• 약학회지
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• 제46권6호
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• pp.426-432
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• 2002

Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV type 16 and 18) and cervical cancer has been well known. An extract of Saururus chinensis, named as PE-46, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. PE-46 inhibited the proliferation of human cervical cancer cell lines in a dose response manner. PE-46 also inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, PE-46 inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by PE-46. SiHa cells treated with PE-46 induced G0/G1 arrest, resulting in inhibition of growth. Our study showed that the PE-46 can inhibit the cervical carcinomas via both inhibition of bindings between oncogenes and tumor suppressors, and inhibition of G1longrightarrowS transition. PE-46 inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus could be used as a putative modulating agent for the treatment of cervical carcinomas caused by HPV.

• 생명과학회지
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• 제22권8호
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• pp.1092-1098
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• 2012

본 연구에서는 우방자를 70% 에탄올로 추출하여 얻어진 추출물을 n-hexane, methylene chloride, ethyl acetate로 순차용매 분획하였다. 각 분획물에 대해 activity-guided isolation을 수행하여 활성물질의 분리 정제를 실시하였다. 활성을 나타내는 물질은 silica gel chromatography (230 mesh), sephadex LH 20, recrystallization method를 이용하여 분리하였다. 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석하였고 arctiin, arctigenin, diarctigenin, matairesinol 으로 동정하였다. 이들을 human dermal fibroblast HS68 세포에 처리하여 얻어진 상등액은 ELISA kit를 활용하여 procollagen type I 생합성과 MMP-1 저해활성을 측정하였다. 측정결과 procollagen type I 생합성과 MMP-1 저해활성 결과 모두 arctiin이 가장 우수한 결과를 보였다. 이와 같은 결과를 통해 우방자에서 분리한 리그난 화합물들을 이용하여 주름개선 소재로 개발할 수 있을 것으로 사료 된다.

• 한방안이비인후피부과학회지
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• 제26권2호
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• pp.45-57
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• 2013

Objectives : Daucus carota sativa has been frequently used as food supplements in many of the Asian countries, and a nutritional medical drug in traditional medicine. This research investigated the effects of Daucus carota sativa extract (DCE) on acute phases of inflammation in Raw 264.7 cells treated with lipopolysaccharide (LPS) in terms of the inhibition of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines production. Methods : NO, $PGE_2$, tumor necrosis factor (TNF)-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6 contents were assayed by ELISA, and expressions of inflammation-related proteins such as inducible NO synthase (iNOS) were determined by immunoblot analyses. Results : DCE treatment attenuated the LPS ability to increase the productions of NO and $PGE_2$ as well as the protein level of iNOS in a concentration-dependent manner. Consistently, treatment of the cells with DCE suppressed the production of TNF-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6. DCE also caused decreases of inhibitor of ${\kappa}B{\alpha}$ phosphorylation induced by LPS in the cells, which means DCE inhibition of NF-${\kappa}B$ activity. Furthermore, DCE blocked LPS-induced phosphorylation of p38 and SAPK/JNK. Conclusion : This study showing here may be of help to understand the action mechanism of DCE, and provide the information for the medical use of Daucus carota sativa for the inflammatory disease.

• Clinical and Experimental Pediatrics
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• 제45권7호
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• pp.875-883
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• 2002

목 적 : 기관지 천식은 Th2 반응 우세로 인한 기도의 호산구증과 IgE 증가를 특징으로 하는 질환이다. 박테리아 DNA 혹은 합성 ODNs인 unmethylated CpG motifs(CpG)은 Th1 사이토카인인 $INF-{\gamma}$와 IL-12 유도와 Th2 사이토카인을 하강 조절하여 기도 내의 호산구증 및 전신적 IgE 생성을 억제하며, 메타콜린 흡입에 의한 기관지 과민성을 방지한다. 저자들은 기도 염증을 유발한 생쥐의 비강내에 합성 CpG ODNs의 점적 투여로 기도의 호산구증과 그 외 면역학적 기전에 대한 영향을 연구하고자 한다. 방 법 : BALB/c 생쥐에 주당 2번씩 면역보강제 없이 OVA를 $10{\mu}g$을 피하 주사하여 감작 시켰다. 10주째에 $50{\mu}g$ OVA를 백서의 비강내로 주입하여 기도 염증을 유도한 후, 4주 뒤인 14주째 다시 생쥐의 비강내에 OVA＋CpG ODNs과 OVA를 점적 흡입 주입하였다. 마지막 항원 유발(14주) 후 4일째에 기관지 폐포 세척액을 추출하여 세포를 염색한 후 호산구를 측정하였고 동시에 비장 세포를 배양하여 사이토카인 측정을 하였다. 비장 세포에서 OVA에 의해 생성된 사이토카인 즉 Th1($INF-{\gamma}$, IL-12), Th2(IL-4, IL-5, IL-13) 사이토카인을 ELISA에 의해 측정하였다. 혈청 항원 특이 IgE, IgG2a 또한 ELISA에 의해 측정하였다. 결 과 : 1) 기관지폐포 세척액의 호중구수 : CpG ODNs을 투여한 군이 7.6[5.8-9.4]%, CpG ODNs을 투여하지 않은 군이 42.0[38.2-44.6]%로 기도 염증 세포내의 호산구증을 현저히 억제시켰다(P<0.01). 2) 혈청 OVA-specific IgE와 IgG2a : CpG ODNs으로 투여한 군과 투여하지 않은 군에서 측정한 혈청 OVA-specific IgE와 IgG2a는 음성 대조군에 비해 현저히 증가하였고, CpG ODNs을 비강내 투여 후 4일째 측정한 혈청 OVA-specific IgE와 IgG2a 수치는 CpG ODNs 투여군과 비투여군 사이에 유의한 차이가 없었다. 3) 사이토카인 분석 : CpG ODNs으로 투여한 군은 투여하지 않은 군에 비해 IL-4, IL-5, IL-13의 생성이 감소하였고 IL-12 생성은 증가하였으나, $INF-{\gamma}$의 상향조절은 보여주지 못하였다. 결 론: CpG ODNs의 백신 접종은 면역반응을 조절하여 기도 염증이 형성된 생쥐에서 기도 호산구증을 감소시키는데 매우 효과적이며 천식 치료에 유용할 수 있을 것으로 사료된다.

• 한국동물위생학회지
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• 제28권2호
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• pp.121-146
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• 2005

Newcastle disease (ND), infectious bronchitis (IB), low pathogenic avian Influenza (LPAI) and fowl typhoid (FT) have been known as egg drop laying diseases because of the serious layer damage from mass zone layer. In this study, such egg drop laying diseases were investigated. To access this study, we peformed to evaluate antibody titers in serum and isolated bacteria and virus from organs and feces on May, July and September in 2003. The distribution of ND from January to May, IB and LPAI from October to February of the next year, and FT from March to September were inspected by the question survey in 21 farms. ND revealed to be positive rates of 490 to 474 $(96.7\%)$ in May, 510 to 506 $(99.2\%)$ in July and 510 to 510 $(100\%)$ in September with hemagglutination inhibition (HI) test. The mean antibody titers were 10.2, 9.9 and 10.2, respectively. With regard to IB, 484 out of 490 samples $(98.7\%)$ in May, 508 of 510 $(99.6\%)$ in July and 509 of 510 $(99.8\%)$ in September showed positive results and the mean antibody titers were gradually increased with 8.2, 9.0 and 9.4, respectively. According to HI test of LPAI, the positive results were shown in 442 of 480 $(92.1\%)$, 394 of 494 $(79.8\%)$ and 402 of 483 $(83.2\%)$ in May, July and September, respectively The mean antibody titers were decreased with 4.6, 4.3 and 4.0. The distribution of LPAI also elicited the positive rates of 480 to 475 $(99.0\%)$ in May, 494 to 485$(98.2\%)$ in July, 483 to 472 $(97.7\%)$ in September as determined by ELISA and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. Compared ELISA results with HI test of LPAI the positive results were 480 to 422 $(92.1\%),\;475(99.0\%),\;494\;to\;394 (79.8\%),\;485 (98.2\%)\;and\;483\;to\;402(83.2\%),\;472(97.7\%)$. Therefore, the positive rate determined by ELISA was higher than that of HI test with 6.9, 18.4 and $14.5\%$, respectively. When performed RT-PCR for ND using organ and feces samples, the pathotypes were detected $5(15.6\%)\;in\;May,\; 2(5.3\%) in\;July,\;2(7.1\%)$ in September but there is no samples showing positive band for LPAI. In attempt to isolate Salmonella gallinarum, bacteria were obtained from 4 cases (12.5%) in May, 9 (23.6%) in July, 5 (17.8%) in September. Thus the highest rate for isolation revealed to be shown in July When evaluated the antimicrobial susceptibility to 18 isolated strains of 5. gallinarum, bacteria were sensitive to trimethoprim/sulfamethox$(61.1\%),\;kanamycin\;(55.5\%),\;ampicillin\;(55.5\%)$ and amoxacillin/clavulanic acid $(55.5\%)$, cephalothin $(50.0\%)$, but resistant to penicillin $(88.9\%)$, streptomycin $(88.9\%)$, erythromycin $(83_4\%)$ and tetracycline $(61.1\%)$. According to HI test of ND and LPAI using captured 164 wild Korean tree sparrows (Passer nontanus), the positive rates were $47.6\%\;and\;57.3\%$, and the mean HI titers were 5.32 and 4.02, respectively. 71 $(43.2\%)\;and\;58(35.3\%)$ in captured sparrows also showed more than 4 titers for HI test to ND and LPAI, respectively However, the attempt for isolation of viruses failed in all samples.

• 대한본초학회지
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• 제23권2호
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• pp.87-97
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• 2008

Objectives : The present study was carried out to investigate the effect of Herba Polygalae Japonica on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mice by ovalbumin (OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dessiciated with collagenase (1 ${\mu}g$/ml). Eosinophils were activated by rIL-3/rmIL-5 co-treatments. The lung cells were treated with extract of Herba Polygalae Japonica (EPJ), incubated for 48 hr at $37^{\circ}C$, and analyzed by flow cytometer, ELISA, RT-PCR and immunocytochemistry stain. Results : A significant cytotoxicity by drug treatment was not observed. The cell number ratio of granulocyte, CD3e-/CCR3+, CD3e+CD69+, CD4+, CD23+/B220+ cells was increased in rmIL-5/rIL-3 treated control group compared to the normal group. Cells numbers in the experimental animal group treated with EPJ was all decreased. In ELISA analysis, IL-4, IL-5, IL-13 levels and histamine release level were increased in the control group compared to the normal animal group, then significantly decreased in the experimental group with 100 ${\mu}g$/ml of EPJ treatment. In RT-PCR analysis, mRNA expressions of IL-4, IL-5, IL-13, CCR3 and eotaxin were increased in the control group compared to the normal animal group, then decreased in the experimental group with 100 ${\mu}g$/ml of EPJ treatment. And eosinophil proliferation levels were 18847${\pm}$,1527 (cpm) in the control group, 4676${\pm}$972 (cpm) in the positive control group, and 7709 ${\pm}$ 549 (cpm), 16839 ${\pm}$ 1403 (cpm), 16385 ${\pm}$ 1723 (cpm) in the experimental group with 100 ${\mu}g$/ml, 10 ${\mu}g$/ml, 1 ${\mu}g$/ml of EPJ treatment. Conclusions : The present data suggested that Herba polygalae japonica may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of EPJ.

• IMMUNE NETWORK
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• 제7권1호
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• pp.39-47
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• 2007

Background: Stromal cell-derived factor (SDF)-1 is a potent chemoattractant for activated T cells into the inflamed Rheumatoid arthritis (RA) synovium. To determine the effect of macrophage migration inhibitory factor (MIF) on the production of SDF-1 in the inflamed RA synovium. Methods: The expression of SDF-1 and MIF in RA and Osteoarthritis (OA) synovium was examined by immunohistochemical staining. The SDF-1 was quantified by RT-PCR and ELISA after RA fibroblast like synoviocyte (FLS) were treated with MIF in the presence and absence of inhibitors of intracellular signal molecules. The synovial fluid (SF) and serum levels of MIF and SDF-1 in RA, OA and healthy control were measured by ELISA. Results: Expression of SDF-1 and MIF in synovium was higher in RA patients than in OA patients. The production of SDF-1 was enhanced in RA FLS by MIF stimulation. Such effect of MIF was blocked by the inhibitors of NF-${\kappa}B$. Concentrations of SDF-1 in the serum and SF were higher in RA patients than in OA patients and healthy control. SDF-1 and MIF was overexpressed in RA FLS, and MIF could up-regulate the production of SDF-1 in RA FLS via NF-${\kappa}B$-mediated pathways. Conclusion: These results suggest that an inhibition of interaction between MIF from T cells and SDF-1 of FLS may provide a new therapeutic approach in the treatment of RA.