• 한국미생물·생명공학회지
• /
• 제38권2호
• /
• pp.198-206
• /
• 2010

본 연구는 항생제인 imipenem에 내성이 있는 Pseudomonas aeruginosa에 대한 차 폴리페놀(TPP)과 imipenem의 살균 상승효과와 세포반응을 조사하기 위하여 수행되었다. Imipenem 내성 Ps. aeruginosa는 병원의 환자로부터 분리하였다. TPP와 imipenem을 단독으로 처리하였을 때와 병용으로 처리하였을 때의 최소억제농도(MIC)를 측정한 결과, imipenem 감수성과 내성 균주는 TPP와 imipenem을 병용처리 하였을 때, imipenem 농도가 각각 16배, 8배가 감소되는 것을 확인하였다. 또한, time-kill 조사를 통해 TPP와 imipenem의 항균효과를 조사하였으며, 병용처리 하였을 때 낮은 농도의 imipenem에서도 동일한 항균효과를 나타내는 것을 확인하였다. TPP에 의한 imipenem 감수성과 내성 균주의 스트레스 충격 단백질 발현을 조사하기 위하여 anti-DnaK와 anti-GroEL 단일항체를 이용한 Western blot을 통해 관찰하였다. 스트레스 충격 단백질인 DnaK와 GroEL은 TPP의 노출시간이 증가함에 따라 발현양이 증가하다가 감소하는 것을 확인하였으며, 유도된 DnaK와 GroEL의 분자량은 각각 70 kDa과 60 kDa으로 나타났다. TPP의 농도와 시간에 따른 세균의 LPS 증감 변화를 SDS-PAGE와 은 염색을 통하여 확인하였고, TPP와 imipenem에 노출된 세균의 세포 외부 형태변화를 주사전자현미경을 이용하여 관찰한 결과, 움푹 패이고, 주름진 표면을 가지는 것으로 관찰되었다.

• Journal of Microbiology and Biotechnology
• /
• 제27권9호
• /
• pp.1628-1638
• /
• 2017

Viola tianshanica Maxim, belonging to the Violaceae plant family, is traditionally used in Uighur medicine for treating pneumonia, headache, and fever. There is, however, a lack of basic understanding of its pharmacological activities. This study was designed to observe the effects of the ethanol extract (TSM) from Viola tianshanica Maxim on the inflammation response in acute lung injury (ALI) induced by LPS and the possible underlying mechanisms. We found that TSM (200 and 500 mg/kg) significantly decreased inflammatory cytokine production and the number of inflammatory cells, including macrophages and neutrophils, in bronchoalveolar lavage fluid. TSM also markedly inhibited the lung wet-to-dry ratio and alleviated pathological changes in lung tissues. In vitro, after TSM ($12.5-100{\mu}g/ml$) treatment to RAW 264.7 cells for 1 h, LPS ($1{\mu}g/ml$) was added and the cells were further incubated for 24 h. TSM dose-dependently inhibited the levels of proinflammatory cytokines, such as NO, $PGE_2$, $TNF-{\alpha}$, IL-6, and $IL-1{\beta}$, and remarkably decreased the protein and mRNA expression of $TNF-{\alpha}$ and IL-6 in LPS-stimulated RAW 264.7 cells. TSM also suppressed protein expression of $p-I{\kappa}Ba$ and p-ERK1/2 and blocked nuclear translocation of $NF-{\kappa}B$ p65. The results indicate that TSM exerts anti-inflammatory effects related with inhibition on $NF-{\kappa}B$ and MAPK (p-ERK1/2) signaling pathways. In conclusion, our data demonstrate that TSM might be a potential agent for the treatment of ALI.

• The Korean Journal of Physiology and Pharmacology
• /
• 제5권1호
• /
• pp.55-63
• /
• 2001

In macrophages, lipopolysaccharide (LPS) alone or in combination with $interferon-{\gamma}\;(IFN-{\gamma})$ has been shown to release a nitric oxide (NO) through the increase of the transcription of the inducible nitric oxide synthase (iNOS) gene. To investigate the exact intracellular signaling pathway of the regulation of iNOS gene transcription by LPS plus $IFN-{\gamma},$ the effects of protein tyrosine kinase (PTK) inhibitor and protein kinase C (PKC) inhibitors on NO production, iNOS mRNA expression, nuclear $factor-_{\kappa}B\;(NF-_{\kappa}B)$ binding activity and the promoter activity of iNOS gene containing two $NF-_{\kappa}B$ sites have been examined in a mouse macrophage RAW 264.7 cells. LPS or $IFN-{\gamma}$ stimulated NO production, and their effect was enhanced synergistically by mixture of LPS and $IFN-{\gamma}.$ The PTK inhibitor such as tyrphostin reduced LPS plus $IFN-{\gamma}-induced$ NO production, iNOS mRNA expression and $NF-_{\kappa}B$ binding activity. In contrast, PKC inhibitors such as H-7, Ro-318220 and staurosporine did not show any effect on them. In addition, transfection of RAW 264.7 cells with iNOS promoter linked to a CAT reporter gene revealed that tyrphostin inhibited the iNOS promoter activity through the $NF-_{\kappa}B$ binding site, whereas PKC inhibitors did not. Taken together, these suggest that PTK, but not PKC pathway, is involved in the regulation of the iNOS gene transcription through the $NF-_{\kappa}B$ sites of iNOS promoter in RAW 264.7 macrophages by LPS plus $IFN-{\gamma}$.

• 한국균학회지
• /
• 제46권2호
• /
• pp.161-176
• /
• 2018

진균류 유래의 ${\beta}$-glucan은 pathogen-associated molecular patterns의 일종이기도 하며 면역촉진과 항암작용을 나타냄이 알려져 있지만 세포 내 신호전달에 관해서는 알려진 바가 많지 않다. 대식세포주인 RAW264.7 세포에 불로초에서 추출한 ${\beta}$-glucan을 처리하였을 때 세포막에서는 덱틴-1, toll-like receptor 2, 4, 6의 발현이 증가되었으며, 세포 내에서는 macrophage inflammatory protein (MIP)-1a, MIP-$1{\beta}$, MIP-$1{\gamma}$, IL-$1{\beta}$ 그리고 tumor necrosis factor (TNF)-${\alpha}$의 발현이 증가되었다. 또한 대식세포주에 불로초의 ${\beta}$-glucan과 PI3K 또는 MEK1/MEK2 억제제를 각각 처리하였을 때에 세포 내의 MIP-1a, MIP-$1{\beta}$, MIP-$1{\gamma}$, interleukin-$1{\beta}$, TNF-${\alpha}$의 발현이 감소되었다. 따라서 불로초의 ${\beta}$-glucan은 대식세포에서 MyD88의 경로인 PI3K/Akt를 경유할 뿐만 아니라 MEK 경로를 활성화시킴으로써 다양한 면역조절작용이 가능한 것으로 여겨진다.

• 한국식품과학회지
• /
• 제50권6호
• /
• pp.642-652
• /
• 2018

본 연구는 LPS 처리 큰포식 세포에서 옻 추출물, 옻 첨가 된장 및 간장 추출물의 항염증 및 산화방지 효과를 확인하였다. 염증 반응은 자극이 가해지면 히스타민, 세로토닌, 프로스타글란딘과 같은 혈관 활성물질에 의해 혈관 투과성이 증대되어 염증을 유발하고 사이토카인, 활성산소종, lysosomal enzyme 등 다양한 매개 인자가 관여한다. 자극에 의한 큰포식세포의 염증반응은 $TNF-{\alpha}$, IL-6, $IL-1{\beta}$와 같은 pro-inflammatory cytokine의 발현이 유도되고, iNOS와 COX-2에 영향을 받는 유전자의 발현을 자극하게 되어 NO 및 $PGE_2$ 등의 염증 인자가 생성된다. 이에 따라 옻 추출물, 옻 첨가 된장 및 간장 추추물의 염증 및 산화방지시스템 관련 유전자 발현을 분석하였다. 그 결과 옻 추출물은 LPS 자극에 의해 생성된 NO, 염증성 사이토카인 및 $PGE_2$의 생성을 유의적으로 감소시켰다. 옻 추출물은 산화방지관련 핵 내 전사인자인 Nrf2 및 관련 유전자의 발현에 영향을 미치지 않았다. 옻첨가 된장 및 간장 추출물은 NO 및 염증성 사이토카인의 생성을 억제하였지만, 염증 및 산화방지관련 유전자의 발현에 영향을 미치지는 않았다.

• 환경생물
• /
• 제36권4호
• /
• pp.601-609
• /
• 2018

본 연구에서는 CyanoHABs를 제어하기 위해 주요 우점종인 Microcystis aeruginosa에 대한 항균 펩타이드의 살조 활성을 조사하고, 구조적 특이성을 바탕으로 새로운 M. aeruginosa 제어 펩타이드를 제작하였다. 본 실험에서 CA-MA 유래 펩타이드 CMA1, CMA2와 Helicobacter pylori 유래 펩타이드 HPA3P, HPA3NT3는 처리 48시간 후 각각 67.3, 73.1, 76.7, 69.8%의 최대 살조 효율을 보였다. 또한, 신규 펩타이드 K160242~5는 처리 48시간 후 각각 64.0, 64.1, 66.4, 70.1%의 최대 살조 효율을 보였다. CA-MA 유래 펩타이드는 처리 24시간 이후 세포의 재성장이 관찰되었으므로 세포 표면과의 정전기 인력을 통해 세포를 응집하고 간접적으로 제어하는 것으로 조사되었다. 반면에, 양친매성 펩타이드는 세포의 재성장이 관찰되지 않았으므로 세포 응집과 더불어 세포 내 침투를 통해 직 간접적으로 세포를 제어하는 것으로 추정되었다. 또한, 펩타이드의 살조 기작 및 효율에는 구성 아미노산의 종류, 수, 구조 및 펩타이드의 분자량, 처리 농도 등이 복합적으로 영향을 미치는 것으로 나타났다. 그러나 펩타이드를 이용한 CyanoHABs의 제어 가능성을 제고하기 위해서는 정확한 기작과 관련 인자들의 확인 및 증명이 요구된다.

• Journal of Ginseng Research
• /
• 제43권3호
• /
• pp.377-384
• /
• 2019

Background: Inflammation is widespread in the clinical pathology and closely associated to the progress of many diseases. Triterpenoid saponins as a key group of active ingredients in Panax notoginseng (Burk.) F.H. Chen were demonstrated to show antiinflammatory effects. However, the chemical structures of saponins in the leaves and stems of Panax notoginseng (PNLS) are still not fully clear. Herein, the isolation, purification and further evaluation of the antiinflammatory activity of dammarane-type triterpenoid saponins from PNLS were conducted. Methods: Silica gel and reversed-phase C8 column chromatography were used. Furthermore, preparative HPLC was used as a final purification technique to obtain minor saponins with high purities. MS, NMR experiments, and chemical methods were used in the structural identifications. The antiinflammatory activities of the isolated saponins were assessed by measuring the nitric oxide production in RAW 264.7 cells stimulated by lipopolysaccharides. Real-time reverse transcription polymerase chain reaction was used to measure the gene expressions of inflammation-related gene. Results: Eight new minor dammarane-type triterpene oligoglycosides, namely notoginsenosides LK1-LK8 (1-8) were obtained from PNLS, along with seven known ones. Among the isolated saponins, gypenoside IX significantly suppressed the nitric oxide production and inflammatory cytokines including tumor necrosis $factor-{\alpha}$, interleukin 10, interferon-inducible protein 10 and $interleukin-1{\beta}$. Conclusion: The eight saponins may enrich and expand the chemical library of saponins in Panax genus. Moreover, it is reported for the first time that gypenoside IX showed moderate antiinflammatory activity.

• Asian-Australasian Journal of Animal Sciences
• /
• 제32권8호
• /
• pp.1112-1121
• /
• 2019

Objective: Gram-negative bacteria lipopolysaccharide (LPS) has been reported to be associated with uterine impairment, embryonic resorption, ovarian dysfunction, and follicle retardation. Here, we aimed to investigate the toxic effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Methods: First, we developed an in vitro model to study the response of bovine cumulusoocyte complexes (COCs) to LPS stress. After incubating germinal vesicle COCs in $10{\mu}g/mL$ of LPS, we analyzed the following three aspects: the expression levels of the LPS receptor toll-like receptor 4 (TLR4) in COCs, activities of intracellular signaling protein p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor-kappa B (NF-${\kappa}B$); and the concentrations of interleukin (IL)-$1{\beta}$, tumor necrosis factor (TNF)-${\alpha}$, and IL-6. Furthermore, we determined the effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Results: The results revealed that LPS treatment significantly elevated TLR4 mRNA and protein expression levels in COCs. Exposure of COCs to LPS also resulted in a marked increase in activity of the intracellular signaling protein p-p38 MAPK and NF-${\kappa}B$. Furthermore, oocytes cultured in maturation medium containing LPS had significantly higher concentrations of the proinflammatory cytokines IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. LPS exposure significantly decreased the first polar body extrusion rate. The cytoplasmic maturation, characterized by polar body extrusion and distribution of peripheral cortical granules, was significantly impaired in LPS-treated oocytes. Moreover, LPS exposure significantly increased intracellular reactive oxygen species levels and the relative mRNA abundance of the antioxidants thioredoxin (Trx), Trx2, and peroxiredoxin 1 in oocytes. Moreover, the early apoptotic rate and the release of cytochrome C were significantly increased in response to LPS. The cleavage, morula, and blastocyst formation rates were significantly lower in parthenogenetically activated oocytes exposed to LPS, while the incidence of apoptotic nuclei in blastocysts was significantly increased. Conclusion: Together, these results provide an underlying mechanism by which LPS impairs maturation potential in bovine oocytes.

• Journal of Dental Anesthesia and Pain Medicine
• /
• 제22권4호
• /
• pp.277-287
• /
• 2022

Background: Inflammatory dental diseases that occur during pregnancy can cause preterm labor and/or intrauterine growth restriction. Therefore, proactive treatment of dental diseases is necessary during pregnancy. Dexmedetomidine (DEX) is a widely used sedative in the dental field, but research on the effect of DEX on pregnancy is currently insufficient. In this study, we investigated the effects of co-treatment with DEX and lipopolysaccharide (LPS) on inflammatory responses in human amnion-derived WISH cells. Methods: Human amnion-derived WISH cells were treated with 0.001, 0.01, 0.1, and 1 ㎍/mL DEX with 1 ㎍/mL LPS for 24 h. Cytotoxicity of WISH cells was evaluated by 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay. The protein expression of cyclooxygenase-2 (COX-2), prostaglandin E₂ (PGE₂), p38, and nuclear factor kappa B (NF-𝜅B) was examined by western blot analysis. The mRNA expression of pro-inflammatory cytokines such as interleukin (IL)-1𝛽 and tumor necrosis factor (TNF)-𝛼 was analyzed by real-time quantitative polymerase chain reaction. Results: Co-treatment with DEX and LPS showed no cytotoxicity in the WISH cells. The mRNA expression of IL-1𝛽 and TNF-𝛼 decreased after co-treatment with DEX and LPS. DEX and LPS co-treatment decreased the protein expression of COX-2, PGE₂, phospho-p38, and phospho-NF-𝛋B in WISH cells. Conclusion: Co-treatment with DEX and LPS suppressed the expression of COX-2 and PGE₂, as well as pro-inflammatory cytokines such as IL-1𝛽 and TNF-𝛼 in WISH cells. In addition, the anti-inflammatory effect of DEX and LPS co-treatment was mediated by the inhibition of p38/NF-𝜅B activation.

• 한방재활의학과학회지
• /
• 제32권2호
• /
• pp.1-17
• /
• 2022

Objectives This study was conducted to investigate the beta-glucan & ginsenoside content, antioxidant activity, anti-inflammatory effect and safety of herbal medicine mix. Methods The marker compounds contents, antioxidant activity and safety of herbal medicine mix were tested. The contents of beta-glucan and ginsenoside Rg3 were measured, the antioxidant activity was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, anti-inflammatory and a safety test was conducted via single dose toxicity assessment. Results Analyzing the contents of marker compounds showed 362.3 mg/g of beta-glucan, and 0.4184 mg/g of ginsenoside Rg3. In the DPPH free radical scavenging activity, the IC50 of herbal medicine mix, was 0.146%. The scavenging activity of herbal medicine mix was 88.28% activity at 0.5% concentration, and 90.61% activity at 5% concentration. In the lipopolysaccharides (LPS) anti-inflammatory test, the herbal remix showed a significant decrease in tumor necrosis factor-alpha (TNF-𝛼) and interleukin-6 (IL-6) compared to the LPS-induced group. In the single dose toxicity test of herbal medicine mix, a dose of 2,000 mg/kg body weight (BW) was set at its highest capacity and observed after oral administration to female and male rats. No toxicological findings were recognized. It was observed that the resulting lethal dose can be set to 2,000 mg/kg BW or higher for both females and males. Conclusions The results of the experiment on herbal medicine mix showed that the marker compounds contents were beta-glucan and ginsenoside Rg3, that antioxidant activity was observed through the DPPH free radical scavenging activity, anti-inflammatory effect was observed through TNF-𝛼 and IL-6 measurement, and safety was confirmed through the single dose toxicity assessment.