• Journal of Ginseng Research
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• 제41권1호
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• pp.43-51
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• 2017

Background: BIOGF1K, a compound K-rich fraction prepared from the root of Panax ginseng, is widely used for cosmetic purposes in Korea. We investigated the functional mechanisms of the anti-inflammatory and antioxidative activities of BIOGF1K by discovering target enzymes through various molecular studies. Methods: We explored the inhibitory mechanisms of BIOGF1K using lipopolysaccharide-mediated inflammatory responses, reporter gene assays involving overexpression of toll-like receptor adaptor molecules, and immunoblotting analysis. We used the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay to measure the antioxidative activity. We cotransfected adaptor molecules, including the myeloid differentiation primary response gene 88 (MyD88) and Toll/interleukin-receptor domain containing adaptor molecule-inducing interferon-${\beta}$ (TRIF), to measure the activation of nuclear factor (NF)-${\kappa}B$ and interferon regulatory factor 3 (IRF3). Results: BIOGF1K suppressed lipopolysaccharide-triggered NO release in macrophages as well as DPPH-induced electron-donating activity. It also blocked lipopolysaccharide-induced mRNA levels of interferon-${\beta}$ and inducible nitric oxide synthase. Moreover, BIOGF1K diminished the translocation and activation of IRF3 and NF-${\kappa}B$ (p50 and p65). This extract inhibited the upregulation of NF-${\kappa}B$-linked luciferase activity provoked by phorbal-12-myristate-13 acetate as well as MyD88, TRIF, and inhibitor of ${\kappa}B$ ($I{\kappa}B{\alpha}$) kinase ($IKK{\beta}$), and IRF3-mediated luciferase activity induced by TRIF and TANK-binding kinase 1 (TBK1). Finally, BIOGF1K downregulated the NF-${\kappa}B$ pathway by blocking $IKK{\beta}$ and the IRF3 pathway by inhibiting TBK1, according to reporter gene assays, immunoblotting analysis, and an AKT/$IKK{\beta}$/TBK1 overexpression strategy. Conclusion: Overall, our data suggest that the suppression of $IKK{\beta}$ and TBK1, which mediate transcriptional regulation of NF-${\kappa}B$ and IRF3, respectively, may contribute to the broad-spectrum inhibitory activity of BIOGF1K.

• Journal of Microbiology and Biotechnology
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• 제29권7호
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• pp.1155-1164
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• 2019

Lichens contain diverse bioactive secondary metabolites with various chemical and biological properties, which have been widely studied. However, details of the inhibitory mechanisms of their secondary metabolites against influenza A virus (IAV) have not been documented. Here, we investigated the antiviral effect of lichen extracts, obtained from South Korea, against IAV in MDCK cells. Of the lichens tested, Nipponoparmelia laevior (LC24) exhibited the most potent inhibitory effect against IAV infection. LC24 extract significantly increased cell viability, and reduced apoptosis in IAV-infected cells. The LC24 extract also markedly reduced (~ 3.2 log-fold) IAV mRNA expression after 48 h of infection. To understand the antiviral mechanism of LC24 against IAV, proteomic (UPLC-$HDMS^E$) analysis was performed to compare proteome modulation in IAV-infected (V) vs. mock (M) and LC24+IAV (LCV) vs. V cells. Based on Ingenuity Pathway Analysis (IPA), LC24 inhibited IAV infection by modulating several antiviral-related genes and proteins (HSPA4, HSPA5, HSPA8, ANXA1, ANXA2, $HIF-1{\alpha}$, AKT1, MX1, HNRNPH1, HNRNPDL, PDIA3, and VCP) via different signaling pathways, including $HIF-1{\alpha}$ signaling, unfolded protein response, and interferon signaling. These molecules were identified as the specific biomarkers for controlling IAV in vitro and further confirmation of their potential against IAV in vivo is required. Our findings provide a platform for further studies on the application of lichen extracts against IAV.

• KSBB Journal
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• 제21권2호
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• pp.133-139
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• 2006

혈액 내 순환시 안정성 향상과 면역원성의 감소를 위해, rhIFN-${\alpha}$-2a은 N-terminus의 ${\alpha}$-아민기에 mPEG aldehyde를 solid-phase PEGylation 시킨다. CM-Sepharose와 같은 양이온 교환수지가 고체 지지체로 사용되었다. Mono-PEGylate는 양이온 교환 수지에서 unmodified 단백질과 분리되어 용출된다. Site-srecific PEGylation과 mono-PEGylate의 분리가 한 단계의 공정으로 얻어진다는 점은 solid-phase PEGylation의 이점을 뒷받침해준다. 위치 특이성은 peptide digest의 질량 분석과 Edman degradation을 이용한 N-terminal sequencing에 의해 확인하였다. Mono-PEGylate는 항바이러스 활성과 면역원성의 감소를 나타내고, 감소 정도는 결합되는 mPEG의 분자량에 비례한다. Trypsin 저항성과 온도 안정성은 mono-PEGylation에 의해 두드러지게 개선되었다. Solid-phase PEGylation을 통해 종래의 액상 반응에서 나타날 수 있는 재현성 낮은 반응, 부 반응물 생성, 부 반응물 제거 공정 등의 단점을 극복할 수 있었다. 그러나 solid-phase PEGylation의 문제점인 액상 반응에 비교하여 많은 양의 PEG를 사용하여야 한다는 점은 개선되어야 한다.

• 한국동물위생학회지
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• 제37권2호
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• pp.111-122
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• 2014

본 연구는 대나무 분말의 항산화 활성이 비육돈에서 면역조절 효과에 미치는 영향을 구명하기 위해 실시하였으며, 이를 위해 대나무 분말 추출물의 항산화능을 검정하기 위하여, 총페놀 함량, 플라보노이드 함량 및 DHHP, ABST, hydroxyl radical 등의 소거활성능을 알아보았다. 대나무 추출물의 항산화능은 높게 나타났으며, DPPH와 ABTS, HRSA 라디칼 소거능은 12~21%였으며, 추출물의 유효성분으로 총페놀 함량은 171 mg/g이었으며, 플라보노이드 함량은 127. mg/g으로 나타났다. 다음으로 대나무 분말이 함유된 배합사료 급여시 돼지의 면역력에 미치는 영향을 알아보았다. 처리구에는 일반사료를 급여하였고, 시험구는 일반사료에 대나무분말을 1.0%, 2.0% 및 3.0%을 첨가하여 급여하였다. 대나무분말 첨가수준에 따라 혈액의 혈구계수 중 백혈구, 적혈구, 헤마토크리트 함량에서 유의적인 수준의 차이를 나타내지 않았다. 하지만 헤모글로빈 함량이 무처리 대조구에 비교하여 증가하였으며 특히 2~3%의 처리구에서 헤모글로빈 함량은 높게 나타났다. 혈액 내 총단백질, 혈중 알부민의 농도를 분석한 결과 처리구와 무처리구간에 유의적인 차이를 나타내지 않았다. AST, ALT 함량은 대나무 분말 처리군에서 무처리군보다 약간 높게 나타나지만 정상적인 범위내에서의 변화를 보였다. immunoglobulin에서 IgA와 IgG 농도는 무처리구에 비교하여 대나무 분말 처리구에서 높게 나타나고, IgA는 처리구 간의 차이가 나타나지 않았지만 IgG농도에서는 대나무 추출물 2%, 3% 처리구가 1% 처리구에 비해 높은 수준을 나타냈다. TNF-${\alpha}$의 경우 무첨가구와 첨가구 사이의 유의적인 차이가 없었지만, 대나무 분말 첨가구에서 interferon-${\gamma}$의 함량은 모든 첨가구에서 무처리 군보다 유의적으로 높게 나타났다. 특히 2%, 3% 처리구가 1%의 처리구 보다도 높은 interferon-${\gamma}$의 함량을 나타내었다. 이는 대나무의 페놀, 또는 플라보노이드 성분에 기초한 높은 항산화력에 의하거나, 또는 대나무에 포함된 돼지 초기 면역반응을 활성화시킬 수 있는 물질에 의한 면역활성 증강에 따른 것으로 사료된다. 결론적으로 대나무 분말의 높은 항산화력은 돼지 면역력 향상에 기여하였으며, 특히 2~3% 대나무 분말 첨가 사료는 돼지 생리활성을 크게 증강시키는 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권2호
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• pp.237-244
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• 2008

The object of this trial was to investigate the effect of dietary ${\beta}$-1,3/1,6-glucan supplementation on the performance and immunological response of broiler chickens. Two hundred and forty 1-day old male broilers ($39{\pm}1g$) were separated into six treatments which were given six different feeds containing 0 (control), 25, 50, 75, 100 and 125 mg/kg dietary ${\beta}$-1,3/1,6-glucan supplementation. On days 21 and 42, body weight gain, feed consumption and feed conversation rate were recorded as measures of growth performance. The levels of key cytokines in the immuno-regulating pathway: interleukin-1 (IL-1), interleukin-2 (IL-2), interferon $\gamma$(IFN-$\gamma$, tumor necrosis factor $\alpha$(TNF-$\alpha$, and the concentrations of signal molecules: peripheral blood plasma globulin, serum Immunoglobulin G (IgG) and intestinal secretary Immunoglobulin A (sIgA), were measured as indices of the immune response to determine suitable levels of dietary ${\beta}$-1,3/1,6-glucan supplementation. The results indicated that performance was elevated quadratically with dietary ${\beta}$-1,3/1,6-glucan supplementation. Maximal growth performance and an enhanced immunological response were obtained at a supplemented level of 50 mg/kg.

• Journal of Nutrition and Health
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• 제24권2호
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• pp.97-103
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• 1991

에탄올이 면역계의 항원전달세포이며, 동시에 감마인터페론 등의 림포카인에 의해 활성화되면 작동세포의 역활을 하는 단핵구-대식세포의 arachildonic acid 대사의 cyclooxygenasc 경로의 프로스타글라딘 생산에 미치는 영향을 연구하기 위하여 감마 인터페론으로 전처치하여 활성화시킨 단핵구와 대조군 단핵구를 사용하여 방사면역측정법으로 프로스타글라딘들을 측정하였다. 건강한 성인의 말초혈액으로 Ficoll-Hypaque용액을 이용한 농도구배법과 젤라틴 피복 플라스크를 이용하여 단핵구를 순수하게 분리한 다음 800단위의 감마 인터페론을 사용하여 3일동안 단핵구를 활성화시켰다. 감마인터페론으로 활성화 시키지 않은 $10^6$개의 단핵구에 100mM,300mM,600mM의 에탄올을 30분간 반응시킨 후 6시간후에 thromboxane B$_2$, PGEB$_2$ 와 6-keto-PGF1$\alpha$ 와 PGE$_2$ 모두 대조군에 비하여 생산이 증가하였다. 감마인터페론으로 전처치한 경우에는 알코올 처치에 관계없이 세가지 프로스타글란딘들 모두 6시간과 24시간에서 대조군보다 감소되었다. 이와같은 성적들로 미루어 볼 때 감마인터페론 처치가 에탄올로 유발한 단핵구 arachidonic acid 대사의 변화를 억제한다고 사료된다.

• 약학회지
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• 제43권2호
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• pp.198-207
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• 1999

The aim of the present investigation was to examine whether YH439, a hepatoprotective agent, exerts protective effect against hepatotoxicity and reduces the production of cytokines and NO in lipopolysaccharide (LPS)-primed rats with carbon tetrachloride ($CCl_4$). Administration of LPS following a single dose of CCl4 injection resulted in remarkable elevations of the serum $TNF{\alpha},{\;}IL-l{\beta$ and IL-6 level. The serum NO level was moderately elevated and severe liver damage was evidenced by increases in serum alanine aminotransferase (ALT) and sorbitol dehydrogenase (SDH) activities. YH439 decreased the levels of TNF, $IL-l{\beta}$, IL-6, ALT, SDH as well as NO in the serum elevated by CCl4+LPS in a dose-dependent manner. Inducible nitric oxide synthase (iNOS) level was decreased in the liver of rats treated with YH439. The increased iNOS activity induced by LPS and $interferon-{\gamma}$ was significantly decreased in RAW 264.7 cells by YH439 treatment. YH439 increased the GSH level decreased by $CCl_4+LPS$ and suppressed the ratio of GSSG/GSH. The reduction of hepatotoxicity by YH439 may associated with the decrease in the production of cytokines as well as suppression of iNOS protein in conjunction with an increase in the GSH level.

• 대한한방내과학회지
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• 제38권6호
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• pp.863-876
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• 2017

Objectives: The aim of this experimental study was to evaluate the anti-neoplastic and anti-inflammatory effects of single and mixed extracts of Ulmus davidiana (UD) and Oldenlandia diffusa (OD) on azoxymethane/dextran sodium sulfate (AOM/DSS)-induced colonic neoplasms in mice. Methods: AOM/DSS induces colitis-associated colonic neoplasms in mice. Mice were divided into seven groups: normal-no inducement and no treatment; control-colonic neoplasms with no treatment; UD-colonic neoplasms and treatment with UD; OD-colonic neoplasms and treatment with OD; UD1+OD1-colonic neoplasms and treatment with UD1 and OD1. UD1+OD2-colonic neoplasms and treatment with UD1 and OD2; UD2+OD1-colonic neoplasms and treatment with UD2 and OD1. Single and mixed preparations of UD and OD were applied to mice for six weeks. The colon length and weight and histopathologic changes of colon tissue were observed. Serum pro-inflammatory cytokines, including tumor necrosis factor-alpha ($TNF-{\alpha}$) and interleukin-6 (IL-6), were determined by enzyme-linked immunosorbent assay. The mRNA expression levels of Bax, Bcl-2, and interferon-gamma ($INF-{\gamma}$) were measured by RT-PCR. Results: The colon length was significantly increased in OD, UD1+OD2, and UD2+OD1 mice, and the colon weight was significantly decreased in OD and UD1+OD2 mice. The morphological change of colon epithelial cells was more suppressed in complex-treatment groups than in single-treatment groups. The inhibitory effect on inflammatory cell invasion was especially shown in UD1+OD2 mice. The serum level of the pro-inflammatory $TNF-{\alpha}$ was decreased in all complex-treatment groups, and the IL-6 level was decreased in UD1+OD1 mice. Single-treatment groups had an increase in the mRNA expression of the pro-apoptosis regulator Bax, and UD2+OD1 decreased the mRNA expression of the anti-apoptosis regulator Bcl-2. The mRNA expression of $INF-{\gamma}$ associated with inflammation was decreased in OD and UD1+OD2 mice. Conclusions: This study suggests that single and mixed extracts of Ulmus davidiana and Oldenlandia diffusa have anti-neoplastic and anti-inflammatory effects on AOM/DSS-induced colonic neoplasms in mice. Therefore, we conclude that UD, OD, and a mixture of UD and OD are potential therapeutic agents for colitis-associated colonic neoplasms.

• 생약학회지
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• 제43권3호
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• pp.217-223
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• 2012

In the present study, we investigated anti-inflammatory activity of artemisinic acid in HaCaT cells and RAW264.7 cells. Artemisinic acid showed inhibitory activity on macrophage-derived chemokines (MDC) expression, a factor related with atopic dermatitis (AD), in interferon (IFN)-${\gamma}$ and tumor necrosis factor (TNF)-${\alpha}$-stimulated HaCaT cells. In the study on action mechanism, pretreated artemisinic acid reduced the phosphorylation of STAT1 and p38 and the degradation of $I{\kappa}B$ by IFN-${\gamma}$ and TNF-${\alpha}$ stimulations. However, artemisinic acid didn't show the inhibitory activity on LPS-induced inflammatory mediators (NO, $PGE_2$, IL-6) in RAW264.7 cell. These results indicate that artemisinic acid inhibits IFN-${\gamma}$ and TNF-${\alpha}$-induced MDC expression through inhibition of signal factors, STAT1, NF-${\kappa}B$, and p38, in HaCaT keratinocytes.

• 대한수의학회지
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• 제58권1호
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• pp.9-16
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• 2018

A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV. Immune parameters (neutralizing antibodies, $CD3^+CD4^+$, $CD3^+CD8^+$, $CD3^+CD4^+CD8^+$ T-lymphocytes, and PRRSV-specific interferon $(IFN)-{\gamma}$ secreting T-lymphocytes) were compared with infection parameters (macro- and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages ($CD172{\alpha}^+PRRSV-N^+\;PAM$) as well as with plasma and lymphoid tissue viral loads. Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of $CD172{\alpha}^+PRRSV-N^+\;PAM$ (p < 0.05) as well as with macroscopic lung lesion (p < 0.01). Plasma and tissue viral loads had significant (p < 0.05) negative correlations with $CD3^+CD4^+CD8^+$ T-lymphocyte percentage in PBMC. Frequencies of $CD3^+CD8^+$ and $CD3^+CD4^+$ T-lymphocytes in 14-dpb PBMC had significant negative correlations with of lymph node (p = 0.04) and lung (p = 0.002) viral loads. $IFN-{\gamma}$-secreting T-lymphocytes frequency had a significant negative correlation with gross lung lesion severity (p = 0.002). However, neutralizing antibody titers had no significant negative correlation (p > 0.1) with infection parameters. The results indicate that T-lymphocytes contribute to controlling PRRSV replication in young piglets born after in-utero infection.