Lipopolysaccharide from <i>Prevotella nigrescens</i> stimulates osteoclastogenesis in cocultures of bone marrow mononuclear cells and primary osteoblasts

Chung, Y.-H.,Chang, E.-J.,Kim, S.-J.,Kim, H.-H.,Kim, H.-M.,Lee, S.-B.,Ko, J. S. Blackwell Publishing Ltd 2006 Journal of periodontal research Vol.41 No.4

<P>Background and Objective: </P><P>Lipopolysaccharide is thought to be a major virulence factor of pathogens associated with periodontal diseases and is believed to stimulate bone resorption <I>in vivo</I>. Although <I>Prevotella nigrescens</I> has been implicated in periodontitis, its role in osteoclastogenesis has not been reported. In this study, we investigated the effects of lipopolysaccharide from <I>P. nigrescens</I> on the formation of osteoclasts and the production of cytokines related to osteoclast differentiation.</P><P>Material and Methods: </P><P>Mouse bone marrow mononuclear cells were cultured in the presence of macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor &kgr;B ligand (RANKL), with or without lipopolysaccharide. Bone marrow mononuclear cells were also cocultured with calvarial osteoblastic cells in the presence or absence of lipopolysaccharide. Osteoclast formation was determined by tartrate-resistant acid phosphatase cytochemistry. The production of osteoprotegerin (OPG), M-CSF, tumor necrosis factor alpha (TNF-&agr;), transforming growth factor-beta (TGF-&bgr;) and prostaglandin E<SUB>2</SUB> (PGE<SUB>2</SUB>) was determined by enzyme-linked immunosorbent assay (ELISA).</P><P>Results: </P><P><I>P. nigrescens</I> lipopolysaccharide inhibited osteoclast differentiation from bone marrow mononuclear cells cultured in the presence of M-CSF and RANKL. However, in the coculture system, <I>P. nigrescens</I> lipopolysaccharide stimulated osteoclastogenesis. Notably, <I>P. nigrescens</I> lipopolysaccharide decreased OPG production but increased TGF-&bgr; secretion. In addition, treatment with <I>P. nigrescens</I> lipopolysaccharide increased PGE<SUB>2</SUB> production during the late stage of the culture period. There was no difference in M-CSF and TNF-&agr; production.</P><P>Conclusion: </P><P>These results demonstrate that <I>P. nigrescens</I> lipopolysaccharide stimulates osteoclastogenesis in the coculture system by decreasing the production of OPG and increasing the production of TGF-&bgr; and PGE<SUB>2</SUB>. Through the mechanisms involving these factors, <I>P. nigrescens</I> lipopolysaccharide may cause alveolar bone resorption in periodontal diseases.</P>

지모의 수용성 추출물이 생쥐 소교세포에서 Lipopolysaccharide로 유발된 Cyclooxygenase-2와 Inducible Nitric Oxide Synthase 발현에 미치는 영향

윤종태 ( Jong Tae Yun ),송윤경 ( Yun Kyung Song ),임형호 ( Hyung Ho Lim ) 한방재활의학과학회 2006 한방재활의학과학회지 Vol.16 No.4

목적 : 지모는 임상에서 해열, 항염, 진정, 이뇨 그리고 항당뇨 작용을 가지고 있어 한의학에서 청열자음의 목적으로 사용되어 왔다. 본 연 구는 생쥐 BV2 신경교세포에서 lipopolysaccharide에 의해 유발되는 염증유도산물인 PGs와 NO의 신경염증반응에 대한 지모의 효과를 살펴보기 위해 시행하였다. 방법 : 지모의 항염증 효과를 알아보기 위하여 lipopolysaccharide는 24시간 반응시키는 한편, 지모는 lipopolysaccharide 처치 1시간 전에 전처치 한 이후 reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, PGE2 immunoassay, NO detection 등의 분석방법을 사용하였다. 결과 : lipopolysaccharide는 cyclooxygenase-2 와 inducible nitric oxide synthase의 mRNA와 단백질의 발현을 증가시켜 prostaglandin E2 합성과 nitric oxide 생성을 증가시켰다. 지모를 전처치 하였을 때에는 lipopolysaccharide에 의하여 발현이 증가되던 cyclooxygenase-2와 inducible nitric oxide synthase mRNA와 단백질의 발현이 억제되었고 그 결과 prostaglandin E2 의 합성과 nitric oxide의 생성도 억제되었다. 결론 : 본 연구 결과, 지모는 여러 가지 염증성 질환에 진통 및 항소염 작용이 있는 것으로 보여진다.

In vitro treatment of lipopolysaccharide increases invasion of Pasteurella multocida serotype B:2 into bovine aortic endothelial cells

Seng Kar Yap,Zunita Zakaria,Siti Sarah Othman,Abdul Rahman Omar 대한수의학회 2018 Journal of Veterinary Science Vol.19 No.2

Pasteurella multocida serotype B:2 causes hemorrhagic septicemia in cattle and buffalo. The invasion mechanism of the bacterium wheninvading the bloodstream is unclear. This study aimed to characterize the effects of immunomodulatory molecules, namely dexamethasoneand lipopolysaccharide, on the invasion efficiency of P. multocida serotype B:2 toward bovine aortic endothelial cells (BAECs) and theinvolvement of actin microfilaments in the invasion mechanism. The results imply that treatment of BAECs with lipopolysaccharide at 100ng/mL for 24 h significantly increases the intracellular bacteria number per cell (p < 0.01) compared with those in untreated anddexamethasone-treated cells. The lipopolysaccharide-treated cells showed a significant decrease in F-actin expression and an increase inG-actin expression (p < 0.001), indicating actin depolymerization of BAECs. However, no significant differences were detected in theinvasion efficiency and actin filament reorganization between the dexamethasone-treated and untreated cells. Transmission electronmicroscopy showed that P. multocida B:2 resided in a vacuolar compartment of dexamethasone-treated and untreated cells, whereas thebacteria resided in cellular membrane of lipopolysaccharide-treated cells. The results suggest that lipopolysaccharide destabilizes the actinfilaments of BAECs, which could facilitate the invasion of P. multocida B:2 into BAECs.

Morphological features and lipopolysaccharide attachment of coliphages specific to Escherichia coli O157:H7 and to a broad range of E. coli hosts

김은진,이혜인,이주훈,류상렬,박종현 한국응용생명화학회 2016 Applied Biological Chemistry (Appl Biol Chem) Vol.59 No.1

The objective of the present study was to analyze host-phage adsorption of bacteriophages infecting Escherichia coli O157:H7 and the other E. coli strains. Out of 55 coliphage strains, we selected seven coliphages infectious only to 23 E. coli O157 and seven other coliphages of broad specificity to E. coli O157:H7 and other 61 E. coli. Escherichia coli O157-specific phages and the broadly specific phages all belonged to the Siphoviridae and Myoviridae family, respectively. Escherichia coli O157-specific phages infected E. coli O157:H7, but not E. coli O157:H7△rfaL, deletion mutant of O-antigen ligase gene for lipopolysaccharide. Five coliphages among the broadly specific phages infected E. coli O103, but not E. coli O103△rfaG, deletion mutant of the glycosyltransferase gene. E. coli O157:H7-specific phages among Siphoviridae recognized O-antigen of E. coli O157, but the broadly specific coliphages of Myoviridae may recognize O-antigen and/or a part of the lipopolysaccharide core as an adsorption site in various E. coli. The receptor of the two coliphage groups interacts with some part of lipopolysaccharide, and the tail morphology of the coliphages may be related to their adsorption to and recognition of a different part of lipopolysaccharide. In particular, specificity of E. coli O157:H7-specific phages carrying the long tail of Siphoviridae for O-antigen as a receptor seems to be high.

Lipopolysaccharide와 어성초 혼합 추출물의 지질대사 상관성

권륜희,강금석,김인덕,하배진 한국독성학회 2007 Toxicological Research Vol.23 No.4

흰쥐 뇌에서의 Lipopolysaccharide-유도 산화적 스트레스에 대한 OQ21과 Melatonin의 작용

배미경(Mee Kyung Bae),최신규(Shinkyu Choi),고문정(Moon Jeong Ko),하헌주(Hun Joo Ha),김화정(Hwa Jung Kim) 대한약학회 2005 약학회지 Vol.49 No.4

Lipopolysaccharide (LPS) induces synthesis of several inflammatory cytokines and nitric oxide (NO). NO in brain is involved not only in the regulation of important metabolic pathways via intracellular cyclic GMP-dependent pathways, but also in neurotoxic damage by reacting with superoxide ion leading to form peroxynitrite radical. Oxidative stress has suggested to be related to the inhibition of NO synthase/cyclic GMP pathway. OQ21 is a new fluorinated quinone com- pound that is recently known to have inhibitory effects on both NO synthase (NOS) and guanylyl cyclase (GC). In this study, we examined effects of OQ21, other known NOS or GC inhibitors, or an antioxidant, melatonin, on the oxidative stress pro- duced by LPS in rat brain. Oxidative stress was observed by using the 2',7'-dichlorofluorescin diacetate to measure intracellular reactive oxygen species (ROS) production and by measuring the formation of thiobarbituric acid reactive substances to measure lipid peroxidation. LPS induced significant increase in both ROS produdction and lipid peroxidation in all brain regions tested (striatum, hippocampus and cortex), which were dissected 6 hr after intraperitoneal administration of LPS to rats. Direct striatal injection of two NOS inhibitors, N-nitro-L-arginine methyl ester and diphenyleneiodonium, or a GC inhibitor, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one, produced no significant RO S increase. However, OQ21 enhanced ROS formation in striatal tissues from LPS-treated rats. Melatonin decreased LPS-induced ROS formation and decreased ROS formation increased by OQ21 in striatum of LPS-treated rats.

Wistar-Kyoto Rat와 Spontaneously Hypertensive Rat, 적출 대동맥에서 Lipopolysaccharide가 Phenylephrine에 의한 수축에 미치는 효과

한형수,성영호,김중영 慶北大學校 醫科大學 1994 慶北醫大雜誌 Vol.35 No.4

Lipopolysaccharide로 산화 스트레스를 유도한 Mouse에서 사보환(四補丸)의 Peroxynitrite 억제 효과

권율,정지천,Kweon, Youl,Jeong, Ji-Cheon 대한한방내과학회 2007 大韓韓方內科學會誌 Vol.28 No.1

Objectives : Peroxynitrite (ONOO-), superoxide anion radical (?O2-) and nitric oxide (NO) are cytotoxic because they can oxidize several cellular components such as proteins, lipids and DNA. They have been implicated in the aging process, and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate Sabohwan's activity for scavenging ONOO- and its precursors. NO and ?02-. Methods : For this study, the fluorescent probes, namely 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4.5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Results : Sabohwanblocked tert-butylhydroperoxide (t-BHP)-induced cell death in a dose-dependent fashion. It scavenged t-BHP-induced ONOO-, NO and ?O2- in YPEN cells. Sabohwan inhibited the generation of ONOO-, NO and ?O2- in the lipopolysaccharide (LPS)-treated mouse kidney postmitochondria both in vitro and in vivo. The lipid peroxide level increased and glutathione level decreased in the LPS-treated mice, whereas the ones in the Sabohwanadministered group among the LPS-treated mice reversed toward their natural levels. Conclusions : These results suggest that Sabohwanis an effective ONOO-, ?O2- and NO scavenger, and thereby it might have a potential role as a therapy against the aging process and age-related diseases.

Lipopolysaccharide을 이용한 생쥐 중이염모델에서의 Interleukin-10의 역할

이주형,변정아,여상원,김준규,박용수 대한이비인후과학회 2011 대한이비인후과학회지 두경부외과학 Vol.54 No.1

Background and Objectives Interleukin (IL)-10 is a major regulator of innate immunity. It interferes with the production of inflammatory mediators by reacting with polymorphonuclear neutrophils, monocytes and macrophages. In addition, it also up-regulates the ex-pression of molecules that amplify the anti-inflammatory effect of IL-10. This study tested the hypothesis that injection of various concentrations of exogenous IL-10 in the middle ear cleft can inhibit the acute stage of otitis media induced by lipopolysaccharide (LPS). And if so, the level of concentration for inhibition can be determined. Materials and Method IL-10 and/or LPS were injected transtympanically into the middle ear cavity of normal mice. Histopathological measurement of submucosal thickness and num-ber of inflammatory cell infiltration was performed at 24 hours after inoculation of IL-10 and/or LPS. Results At 24 hours after inoculation, every group of IL-10 inoculation reduced both thickening of the mucous and submucosal infiltration of inflammatory cells in a dose-dependent manner. As the concentrations of IL-10 inoculated increased, mucosal thickness and submucosal infiltration of inflammatory cells were more reduced. Conclusion This study demonstrated that exogenous IL-10 injection into the middle ear can directly reduce LPS-induced otitis media in acute stage in a dose-dependent manner. These findings also indicate that IL-10 may be a candidate for local modulator of acute otitis media.

CD11c항체가 Lipopolysaccharide에 의한 국소적 Shwartzman 반응의 유발에 미치는 영향

박소영,강대영 충남대학교 의과대학 지역사회의학연구소 2000 충남의대잡지 Vol.27 No.2

Lipopolysaccharide(LPS), a component of the cell walls of gram negative bacilli, may induce sepsis, disseminated intravascular coagulation, and multisystem organ failures. The, Shwartzman reaction is an experimental model of LPS-inducing inflammation with localized or generalized intravascular coagulation. This study was carried out to investigate the effects of CD11/CD18 blocking with anti-CD11c antibody on induction of the localized Shwartzman reaction. The control group consisted of mice subcutaneously injected with E.coli LPS(0.01mg). The experimental groups were divided into two subgroups: One consisted of the mice intraperitoneally injected with LPS(2.5mg/kg) with a 24 hour interval after subcutaneous injection with LPS. The other group consisted of mice intraperitoneally injected with LPS with a 24 hour interval after intradermal injection with LPS and with pretreatment of anti-CD11c antibody(20ug). The mice were sacrificed at 24 hours after the final injection. The skins were observed by light and electron microscopy. TNF-α mRNA expression of the experimental group was determined by RTPCR. With light microscopy. the skin of the control group revealed moderate infiltration of neutrophils and monocytes in the dermis. The experimental group injected with LPS in the subcutaneous and intrap:ritoneal areas demonstrated diffuse and heavy infiltration of neutrophils and monocytes with focal necrosis and hemorrhage in the dermis. These findings were consistent with the Shwartzman reaction. The mice subcutaneously and intraperitoneally injected LPS with pretreatment of anti-CD11c antibody showed moderate infiltration of neutrophils and monocytes in the dermis without hemorrhage and necrosis. With electron microscopy, the control group showed the accumulation of a fine granular substance, as seen in edema, and infiltration of neutrophils and monocytes. The capillaries revealed mild endothelial edema without fibrin thrombi formation. In the experimental groups, the mice with the induced Shwartzman reaction demonstrated necrotic inflammatory cell debris and accumulation of edema fluid substance in dermal tissues. The capillaries showed swelling, separation and denudation of endothelial cells with formation of microthrombi composed of fibrin and platelets. However, the anti-CD11c antibody pretreated experimental group showed intradermal infiltration of neutrophils and monocytes and edema without necrosis or microthrombi. On RT-PCR, TNF-a mRNA was definitely expressed in dermal tissues of Shwartzman reaction-induced group but was not in anti-CD11c antibody-pretreated group. According to the above results, it can be concluded that the CD11c/CD18 is a LPS receptor and that the LPS-induced Shwartzman reaction is effectively suppressed with the anti-CDllc antibody.