• 미생물학회지
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• 제27권2호
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• pp.147-154
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• 1989

Vibrio vulnificus Lipopolysaccharide (LPS) was extracted, performed chemical analysis, tested its biological activities, and compared to those of Escherichia coli LPS and Salmonella typhimurium LPS. The lethal activity of V. vulnificus LPS was 138.6138.6 mg/kg in mouse, but this was lower than thowe of E. coli LPS (56.3 mg/kg) and S. typhimurium LPS (37.5 mg/kg). The result of fatty acid analysis showed that V. vulnificus LPS had more saturated fatty acid than E. coli LPS and S. typhimurium LPS. Above results indicated that V. vulnificus LPS did not have much effect on the lethality. The results of biological responses of enzymes and blood cells by LPSs showed that V. vulnificus LPS had slightly greater activity than E. coli LPS and S. typhimurium LPS. V. vulnificus LPS was recommendavle for stimulant on interferon induction because of adequate stimulation and safety for host and cell lines.

• 약학회지
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• 제51권1호
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• pp.44-50
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• 2007

Acute septic shock is one of inflammatory diseases mediated by pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. In this study, we examined the pathological difference and mechanism of lipopolysaccharides isolated from E. coli (E-LPS) or S. abortus (S-LPS) on inducing acute septic shock in ICR mouse. All mice were died by intraperitoneal treatment of S-LPS with 0.75 mg/kg, whereas E-LPS treated with even 3 mg/kg only showed 30% of mice lethal, indicating that S-LPS may be more feasible in triggering a strong septic shock condition. The secretion pattern of TNF-${\alpha}$, a critical pro-inflammatory cytokine in septic shock condition, was also distinct between E-LPS- and S-LPS-treated groups. Thus, S-LPS strikingly increased serum level of TNF-${\alpha}$ (6 ng/ml) at 1 h, while E-LPS just displayed at 2 ng/ml level. However the interaction of S-LPS with LPS receptor toll like receptor (TLR)-4, was not stronger than that of E-LPS, according to experiments with macrophage cell line RAW264.7 cells. Thus, E-LPS rather than S-LPS strongly enhanced the production of TNF-${\alpha}$. Interestingly, S-LPS more strongly up-regulated splenocyte proliferation, compared to E-LPS group, whereas there was no difference between S- or E-LPS treated groups in proliferation of Balb/c- or C57BL/6-originated splenic lymphocytes. Therefore, our data suggest that S-LPS is a more active endotoxin and that the strong septic shock-inducing effect of S-LPS seems due to the enhancement of early TNF-${\alpha}$ production and S-LPS-sensitive lymphocyte proliferation.

• Toxicological Research
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• 제34권1호
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• pp.65-73
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• 2018

Pretreatment of low-dose lipopolysaccharide (LPS) induces a hyporesponsive state to subsequent secondary challenge with high-dose LPS in innate immune cells, whereas super-low-dose LPS results in augmented expression of pro-inflammatory cytokines. However, little is known about the difference between super-low-dose and low-dose LPS pretreatments on immune cell-mediated inflammatory and hepatic acute-phase responses to secondary LPS. In the present study, RAW 264.7 cells, EL4 cells, and Hepa-1c1c7 cells were pretreated with super-low-dose LPS (SL-LPS: 50 pg/mL) or low-dose LPS (L-LPS: 50 ng/mL) in fresh complete medium once a day for 2~3 days and then cultured in fresh complete medium for 24 hr or 48 hr in the presence or absence of LPS ($1{\sim}10{\mu}g/mL$) or concanavalin A (Con A). SL-LPS pretreatment strongly enhanced the LPS-induced production of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, TNF-${\alpha}$/IL-10, prostaglandin E2 ($PGE_2$), and nitric oxide (NO) by RAW 264.7 cells compared to the control, whereas L-LPS increased IL-6 and NO production only. SL-LPS strongly augmented the Con A-induced ratios of interferon (IFN)-${\gamma}$/IL-10 in EL4 cells but decreased the LPS-induced ratios of IFN-${\gamma}$/IL-10 compared to the control, while L-LPS decreased the Con A- and LPS-induced ratios of IFN-${\gamma}$/IL-10. SL-LPS enhanced the LPS-induced production of IL-6 by Hepa1c1c-7 cells compared to the control, while L-LPS increased IL-6 but decreased IL-$1{\beta}$ and C reactive protein (CRP) levels. SL-LPS pretreatment strongly enhanced the LPS-induced production of TNF-${\alpha}$, IL-6, IL-10, $PGE_2$, and NO in RAW 264.7 cells, and the IL-6, IL-$1{\beta}$, and CRP levels in Hepa1c1c-7 cells, as well as the ratios of IFN-${\gamma}$/IL-10 in LPS- and Con A-stimulated EL4 cells compared to L-LPS. These findings suggest that pre-conditioning of SL-LPS may contribute to the mortality to secondary infection in sepsis rather than pre-conditioning of L-LPS.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1120-1126
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• 2007

This study was performed to screen probiotic bifidobacteria for their ability to bind and neutralize lipopolysaccharides (LPS) from Escherichia coli and to verify the relationship between LPS-binding ability, cell surface hydrophobicity (CSH), and inhibition of LPS-induced interleukin-8 (IL-8) secretion by HT-29 cells of the various bifidobacterial strains. Ninety bifidobacteria isolates from human feces were assessed for their ability to bind fluorescein isothiocyanate (FITC)-labeled LPS from E. coli. Isolates showing 30-60% binding were designated LPS-high binding (LPS-H) and those with less than 15% binding were designated LPS-low binding (LPS-L). The CSH, autoaggregation (AA), and inhibition of LPS-induced IL-8 release from HT-29 cells of the LPS-H and LPS-L groups were evaluated. Five bifidobacteria strains showed high levels of LPS binding, CSH, AA, and inhibition of IL-8 release. However, statistically significant correlations between LPS binding, CSH, AA, and reduction of IL-8 release were not found. Although we could isolate bifidobacteria with high LPS-binding ability, CSH, AA, and inhibition of IL-8 release, each characteristic should be considered as strain dependent. Bifidobacteria with high LPS binding and inhibition of IL-8 release may be good agents for preventing inflammation by neutralizing Gram-negative endotoxins and improving intestinal health.

• 한국식품영양과학회지
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• 제37권9호
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• pp.1126-1135
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• 2008

본 연구에서는 내독소인 LPS로 산화적 스트레스를 유발시킨 BALB/c mice에 genistein을 투여하였을 때 TNF-$\alpha$, TBARS, superoxide anion 농도와 GSH, 항산화 효소계 활성, 그리고 NF-${\kappa}B$ transactivation에 미치는 영향을 조사하여 genistein이 내독소에 의한 산화적 스트레스와 염증반응을 억제하는 효과가 있는지 알아보고자 하였다. 평균체중 20 g인 BALB/c mice 암컷 120수를 30수씩 완전임의배치로 4군으로 분류하여 PBS군(대조군)은 PBS를 복강 속으로 투여 후 30분경과 후에 다시 PBS를 투여하였으며, genistein군은 genistein을 체중 kg당 200 mg으로 투여한 30분 후 PBS를 투여하였다. LPS군은 LPS를 처리한 군으로 PBS 투여 30분 후 LPS를 체중 kg당 100 mg 농도로 복강 투여하였고, genistein+LPS군은 체중 kg당 genistein 200 mg을 투여 30분 후 LPS를 체중 kg당 100 mg을 투여하였다. 마지막 투여 1시간, 4시간, 8시간 경과 후 mouse의 안와정맥으로부터 혈액을, 복강으로부터 복강대식세포와 간을 취하였다. LPS 투여 8시간 경과 후 LPS군의 ${O_2}^-$ 생성량은 현저하게 증가하였으며 geinstein+LPS군은 genistein 대조군, PBS군과 비슷한 수준을 유지하였다. 반면 SOD 활성은 geinstein+LPS군이 LPS군에 비해 유의적으로 높은 수준이었다. 혈장에서의 TNF-$\alpha$ 수준은 LPS 투여 8시간 후 genstein+LPS군이 LPS군보다 유의적으로 낮은 수준을 보였다. LPS 투여는 항산화 효소계 활성과 GSH의 수준을 감소하였으나, genistein을 투여한 genistein+LPS군은 LPS군에 비해 GSH 농도와 catalase, GSH-px, GSH-reductase 활성이 모두 유의적으로 증가하는 결과를 보였다. 간에서의 NF-${\kappa}B$ transactivation 정도는 LPS 투여 후 1시간, 4시간 경과 후에 PBS군에 비해 LPS군과 genistein+LPS군에서 유의적으로 높은 수준이었으나 8시간 경과 후 LPS군은 증가하는 반면 genistein+LPS군은 변화하지 않았다. 이상의 결과를 요약해보면 LPS의 투여는 혈장과 간의 산화적 스트레스와 염증반응을 촉진하는 것으로 나타났으며 LPS 투여 전 공급한 genistein은 LPS로 유도된 산화적 스트레스와 염증반응을 항산화 효소계 활성 증가와 NF-${\kappa}B$ transactivation 억제, TNF-$\alpha$ 생성 저하 등의 기작으로 세포내의 과산화수준을 수준을 낮추고 GSH를 증가시켜 산화적 스트레스를 억제하는 것으로 사료된다.

• Journal of Animal Science and Technology
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• 제63권1호
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• pp.114-124
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• 2021

The objective of this study was to characterize the enzymatic hydrolysis of lipopolysaccharide (LPS) by wheat phytase and to investigate the effects of wheat phytase-treated LPS on in vitro toxicity, cell viability and release of a pro-inflammatory cytokine, interleukin (IL)-8 by target cells compared with the intact LPS. The phosphatase activity of wheat phytase towards LPS was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine. In vitro toxicity of LPS hydrolyzed with wheat phytase in comparison to intact LPS was assessed. Cell viability in human aortic endothelial (HAE) cells exposed to LPS treated with wheat phytase in comparison to intact LPS was measured. The release of IL-8 in human intestinal epithelial cell line, HT-29 cells applied to LPS treated with wheat phytase in comparison to intact LPS was assayed. Wheat phytase hydrolyzed LPS, resulting in a significant release of inorganic phosphate for 1 h (p < 0.05). Furthermore, the degradation of LPS by wheat phytase was nearly unaffected by the addition of L-phenylalanine, the inhibitor of tissue-specific alkaline phosphatase or L-homoarginine, the inhibitor of tissue-non-specific alkaline phosphatase. Wheat phytase effectively reduced the in vitro toxicity of LPS, resulting in a retention of 63% and 54% of its initial toxicity after 1-3 h of the enzyme reaction, respectively (p < 0.05). Intact LPS decreased the cell viability of HAE cells. However, LPS dephosphorylated by wheat phytase counteracted the inhibitory effect on cell viability. LPS treated with wheat phytase decreased IL-8 secretion from intestinal epithelial cell line, HT-29 cell to 14% (p < 0.05) when compared with intact LPS. In conclusion, wheat phytase is a potential therapeutic candidate and prophylactic agent for control of infections induced by pathogenic Gram-negative bacteria and associated LPS-mediated inflammatory diseases in animal husbandry.

• 한국자원식물학회지
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• 제25권4호
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• pp.482-489
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• 2012

부화 초기의 육계 병아리에게 천화분추출물을 급여한 후, LPS 처리에 의한 염증유도로 천화분추출물이 염증반응에 미치는 영향을 검토했다. 각 처리군 별 혈장 ceruloplasmin 및 ${\alpha}$-산성단백 농도의 경시적 변동경향은 전 처리군 동일하게 LPS 처리 후 증가하였다. 그러나 각 처리군 별 농도는 LPS 처리 후 3h, 9h 및 24h째 모두에서 천화분 추출물의 첨가량이 증가함에 따라 감소하는 경향을 보였다. 혈장 일산화질소 농도는 전 처리군 모두가 LPS 처리 후, 3h째에 급격하게 증가하였다. 각 처리군 별 수치는 LPS 처리 후, 3h 및 9h째 모두에서 천화분 추출물의 첨가량이 증가함에 따라 하락했다. LPS 처리 후, 간장 내의 IL-2 mRNA량은 모든 처리군들이 동일하게 2h째에 급격한 증가 현상을 보였다가, 그 후 점진적으로 하락하여 9h째에는 LPS처리 전 수준으로 회복했다. 각 처리군 별 경시적 변동 경향은 LPS 처리 후, 2h, 3h 및 4h째에서 천화분 첨가군 모두가 대조군보다 낮은 값을 보였다. LPS 처리 후, 비장 내의 IL-2 mRNA 량은 모든 처리군 들이 4h째에서 가장 높은 수치를 보였다. 처리군 별 변동 값은 천화분 0.2% 및 0.3%첨가군들이 LPS 처리 후, 3h 및 4h에서 대조군과 0.1% 천화분 첨가군들보다 높은 수치를 나타내었다. LPS 처리 후, 간장 및 비장내의 IFN-${\gamma}$ mRNA량은 모든 처리군들이 LPS 처리 후, 2h 및 3h째까지 증가하였으나, 4h에서 하락하여, 9h째에서는 LPS 처리 전 수준으로 하락했다. 그러나 각 처리군별 경시적 변동치는 최고치를 나타낸 LPS 처리 후 2h 및 3h에서, 천화분 추출물 첨가량이 증가함에 따라 하락하였다. LPS 처리 후, 간장 및 비장 내의 iNOS mRNA량은 전 처리군이 3h째에서 가장 높은 수치를 나타내었으며, 9h째에서 LPS 처리 전 수준으로 회복되었다. 처리군별 2h째, 3h째 및 4h째의 수치는 천화분 추출물 첨가군이 대조군보다 낮은 수치를 나타내었다.

• Tuberculosis and Respiratory Diseases
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• 제61권4호
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• pp.374-383
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• 2006

연구배경: 급성폐손상에서 활성산소종에 의한 산화 손상은 주요한 역할을 한다. Ethyl pyruvate (EP) 는 체내에서 생성되는 pyruvate의 유도체로 항산화 및 항염증 효과가 있음이 알려졌다. 저자들은 리포다당질에 의한 급성폐손상 모델에서 EP가 염증반응에 미치는 영향을 연구하고자 하였다. 방 법: 5주 령의 BALB/c 생쥐를 이용하여 리포다당질을 기관 내로 투여하여 급성폐손상을 유도하였다. 대조군, LPS군, EP+LPS군, LPS+EP군으로 나누어 기관지폐포세척액에서 TNF-$\alpha$, IL-6 및 myeloperoxidase (MPO)의 활성을, 폐조직에서 급성폐손상 지수와 NF-$\kappa$B의 농도를 측정하였다. 결 과: EP+LPS군에서 TNF-$\alpha$ 및 IL-6의 농도는 LPS군과 비교하여 감소하였고 (p<0.05) 이들 염증성 시토카인의 농도의 변화는 NF-$\kappa$B의 농도의 변화와 상관 관계를 보였다 (p<0.01). 급성폐손상 지수는 EP+LPS군 및 LPS+EP군에서 LPS군과 비교하여 낮았고 (p<0.05) MPO활성은 EP+LPS군에서 LPS군에 비해 낮았다 (p<0.05). 결 론: EP는 LPS로 인한 급성폐손상에 있어서 예방 및 치료 효과가 있는 것으로 판단된다.

• Journal of Microbiology
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• 제34권3호
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• pp.255-262
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• 1996

The lipopolysaccharide (LPS) yields were measured in Rhodobacter capsulatus under several conditions by the ELISA method. The purification of LPS was done by affinity chromatography of IgG coupled CNBr-activated sepharose-4B instead of ultra-centrifugation. The purity of the LPS didn't show much difference between affinity chromatography and ultra-centrifugation method, but affinity chromatography method required much fewer organisms and was more convenient. LPS yield was measured in ng units by the ELISA method. Mannitol was a better single carbon source than other sugars, but mixing two carbon sources resulted in greater LPS yields than any sugar alone. LPS yield was directly proportional to $NH_ 4CI$ concentration, with optimum yields at 0.05% nitrogen. In contrest to LPS yields, which decreased at 0.005% nitrogen concentration total protein was increased 16 times. Calcium influenced LPS yields. At 0.7 mM $CaCI_ 2$, the LPS yield was 16.5 $\mu$g/mg DW, five times the yield without calcium.

• BMB Reports
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• 제50권2호
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• pp.55-57
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• 2017

Toll-like receptor 4 (TLR4) together with MD2, one of the key pattern recognition receptors for a pathogen-associated molecular pattern, activates innate immunity by recognizing lipopolysaccharide (LPS) of Gram-negative bacteria. Although LBP and CD14 catalyze LPS transfer to the TLR4/MD2 complex, the detail mechanisms underlying this dynamic LPS transfer remain elusive. Using negative-stain electron microscopy, we visualized the dynamic intermediate complexes during LPS transfer-LBP/LPS micelles and ternary CD14/LBP/LPS micelle complexes. We also reconstituted the entire cascade of LPS transfer to TLR4/MD2 in a total internal reflection fluorescence (TIRF) microscope for a single molecule fluorescence analysis. These analyses reveal longitudinal LBP binding to the surface of LPS micelles and multi-round binding/unbinding of CD14 to single LBP/LPS micelles via key charged residues on LBP and CD14. Finally, we reveal that a single LPS molecule bound to CD14 is transferred to TLR4/MD2 in a TLR4-dependent manner. These discoveries, which clarify the molecular mechanism of dynamic LPS transfer to TLR4/MD2 via LBP and CD14, provide novel insights into the initiation of innate immune responses.