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.

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.

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.

Lipopolysaccharide로 유도한 RAW 264.7 세포에 대한 Meyerozyma guilliermondii YJ34-2와 Rhodotorula graminis YJ36-1의 항염활성과 Nitric Oxide 생성 저해물질의 생산

배상민 ( Sang-min Bae ),한상민 ( Sang-min Han ),이종수 ( Jong-soo Lee ) 한국균학회 2017 韓國菌學會誌 Vol.45 No.4

The anti-inflammatory effects of cell-free extracts from wild yeasts, Meyerozyma guilliermondii YJ34-2 and Rhodotorula graminis YJ36-1, caused by the inhibition of nitric oxide (NO) activity and cytotoxic effects were determined. Cell-free extracts from these two yeast strains had dose-dependent inhibitory effects on the production of lipopolysaccharide-induced NO and there were no cytotoxic effects on the treated cells or negative effects on their proliferation. Their cell-free extracts were also shown to have inhibitory effects on pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α and prostaglandin-E₂, in a dose-dependent manner. Maximal inhibitory activity on NO production occurred in cell-free extracts of Meyerozyma guilliermondii YJ34-2 cultivated at 30°C for 24 hr and Rhodotorula graminis YJ36-1 cultivated at 25°C for 24 hr in the yeast extract-peptone-dextrose (YPD) media.

Lipopolysaccharide로 자극시킨 방사선 조사 치은 섬유아 세포에서 granulocytemacrophage colony-stimulating factor와 transforming growth factor-β1 생성

김홍식(Hong-Sik Kim),이성근(Seong-Geun Lee),김광혁(Kwang-Hyuk Kim),김욱규(Uk-Kyu Kim),김종렬(Jong-Ryoul Kim),정인교(In-Kyo Chung),양동규(Dong-Kyu Yang) 대한구강악안면외과학회 2002 대한구강악안면외과학회지 Vol.28 No.3

Purpose: Irradiation in the oral cancer patients causes early and late complications such as intraoral mucositis and fibrosis, with a various expression of GM-CSF and TGF-β. The purpose of this study was to investigate the production of GM-CSF and TGF-β1 by the irradiated human gingival fibroblasts cultivated with lipopolysaccharide. Materials and Methods: Irradiated (total dose, 60 Gy) human gingival fibroblasts were incubated with LPS. Culture supernatants that were collected at 24, 48, and 72 hours were assessed for GM-CSF and TGF-β1 by enzyme-linked immunosorbent assay. Results: 1. GM-CSF production in nomal gingival fibroblasts was increased with incubation time, but decreased with incubation time in irradiated gingival fibroblasts. GM-CSF production in both normal and irradiated gingival fibroblasts induced with LPS was higher than the control. 2. TGF-β1 production in normal gingival fibroblasts was decreased after 24 hours, but, it was increased until 48 hours in irradiated gingival fibroblasts. TGF-β1 production in normal gingival fibroblasts exposed with LPS was higher than the control. Conversely, It was lower than the control in irradiated gingival fibroblasts exposed with LPS. Conclusion: This indicates that irradiation in gingival fibroblasts may play an important role in radiation-induced intraoral mucositis and fibrosis. However, LPS decreases the production of TGF-β1 in the irradiated gingival fibroblasts.

Lipopolysaccharide를 투여한 흰쥐 콩팥에서 iNOS 발현과 NADPH-diaphorase 활성의 비교

최재연(Jae youn Choi),이소영(So yeong Lee),차정호(Jung-ho Cha) 대한해부학회 2009 Anatomy & Cell Biology Vol.42 No.4

유도성 산화질소합성효소(inducible nitric oxide synthase, iNOS)는 다양한 생리작용에 관여함으로 많은 연구가 이루어져 있다. 그러나 형태학적 연구보고는 연구자에 따라 매우 달라서 iNOS의 발현에 대한 검토가 필요하다. 본 연구에서는 4개 회사(Chemicon, CH; Sigma, SI; Transduction Laboratorie, TL; Upstate, UP)의 iNOS 항체의 면역염색결과를 비교하고, 모든 아형의 NOS에 대한 염색이 가능한 NADPH-diaphrase에 대한 광학적 및 전자현미경적 효소조직화학법을 병용함으로써 iNOS 발현을 확인하고자 하였다. SD계 흰쥐에 LPS를 투여하고 iNOS의 발현을 유도하였다. LPS 투여군에서 NADPH-d 전자현미경적 효소조직화학법의 결과 반응산물의 위치가 세포질기질에 있는 세포와 사립체에 있는 세포로 구별되었다. 반응산물이 세포질기질에 있는 세포는 치밀반점, 내림가는세관(DTL), 주머니상피(CE) 및 사이질의 유주세포(WC)이며, 대부분 세뇨관의 세포에는 사립체에 반응산물이 위치하였다. LPS 투여군에서 DTL, CE 및 WC가 염색된 항체는 TL과 UP이나, 사립체에 반응산물이 있는 세뇨관세포도 일부 염색되었다. 본 실험의 결과로서 NDAPH-d 효소조직화학법을 병용하면 iNOS를 발현하는 세포를 정확하게 동정할 수 있을 것으로 생각된다. Inducible nitric oxide synthase(iNOS) has been known to be involved in the various physiological metabolim and has been attracting topic. However, there are extensive differences in the reports about the localization of iNOS expression. To resolve this discrepancy, we compared immunohistochemical data from four iNOS antibody produced by different company(Chemicon, CH; Sigma, SIl Transduction Laboratories, TL; Upstate, UP), and NADPH-diaphorase(NADPH-d) enzyme-histochemical results using light- and transmission electorn-microscope in the lipo-polysaccharide(LPS)-treated rat kidney. Electron microscopical examination revealed two different distribution of the NADPH-d reaction product. In the majority of NADPH-d reaction-positive cells, reaction depositions were restricted to the mitochondia, and in the cells of macula densa, descending thin limb(DTL), capsular epithelium(CE) and interstitial wandering cells(WC), NADPH-d positivities were found in the cytoplasm. In immunohistochemical results from LPS-treated animal, DTL, CE and WC were positively stained with TL and UP antibodied but with CH and SI antibodies. We conclude that NADPH-d histochemistry may be usefull for identifing the iNOS-positive cells morphologically.

Lipopolysaccharide로 활성화시킨 흰쥐 혈관의 iNOS 발현에 대한 Higenamine의 효과

강영진,이균우,구의본,이회영,장기철,Kang Young-Jin,Lee Goun-Woo,Ku Eui-Bon,Lee Hoi-Young,Chang Ki-Churl 대한약리학회 1997 The Korean Journal of Physiology & Pharmacology Vol.1 No.3

Higenamine was widely used as traditional remedy for the treatment of rhumatoid arthritis. Nitric oxide(NO) may be a critical mediator in this inflammatory disease. Synovial tissue from humans with inflammatory arthritis expresses NOS2(iNOS) mRNA and protein, and generates NO in vitro. We therefore, investigated the effect of higenamine on the induction of nitric oxide synthase(NOS) promoted by lipopolysaccharide(LPS). Prophylactic application of higenamine selectively prevented LPS-primed initiation of L-arginine-induced relaxation and restored rhenylephrine(PE)-induced contraction in rat aorta. LPS-stimulated nitrite production in the incubation medium was reduced by higenamine. Furthermore, RT-PCR and Northern analysis indicated that higenamine reduced iNOS expression primed by LPS in rat aorta. These results suggest that higenamine prevents LPS-promoted induction of NOS in vascular smooth muscle.

면역억제제가 Lipopolysaccharide에 의한 생쥐의 간 및 뇌조직의 Nitric Oxide Synthase 활성도의 변화에 미치는 영향

민병우(Byung Woo Min),한형수(Hyng Soo Han),박정숙(Jung Sook Park),김중영(Choong Young Kim) 대한약리학회 1995 대한약리학잡지 Vol.31 No.2

To verify the effect of immunosuppressants on the endotoxin-induced increase in iNOS activity, the action of immunosuppressants, dexamethasone (1.5 mg/kg), azathioprine (5 mg/kg/day) and cyclosporine (10 mg/kg), were evaluated in mice pretreated with LPS. The intraperitoneal injection of lipopolysaccharide (10 mg/kg) increased the nitric oxide synthase (NOS) activity in the brain and liver to maximum at 1 and 3 hours, respectively. The increase in NOS activity was blocked by the treatment with NOS inhibitor, LNAME(300 mg/kg) and aminoguanidine(100 mg/kg); a protein inhibitor, cycloheximide (10 mg/kg); and a transcription inhibitor of inducible NOS(iNOS), dexamethasone(1.5 mg/kg). Immunosuppressants, azathioprine (5 mg/kg) and cyclosporine (10 mg/kg), effectively blocked the increase in NOS activity. These results suggest that iNOS expression plays an important role in LPS-induced the increase in NOS activity and that immunosuppressants can be used as candidate for therapeutic agents in endotoxemia.

Lipopolysaccharide에 의해 유발된 뇌 조직의 염증성 시토카인에 대한 Fenofibrate의 보호 효과

박현욱,유수진,박재황 대한노인병학회 2012 Annals of geriatric medicine and research Vol.16 No.4

Background: Increasing evidence suggests that the entry of proinflammatory factors into the brain induces the activation of microglia, astrocytes, and oligodendrocytes and then produces the more proinflammatory cytokines and reactive oxygen species that can ultimately lead to neuronal death. This has clinical implications and provides a link between periperal inflammation and neuroinflammation. Peroxisome proliferator-activated receptors (PPARs), a member of the nuclear hormone receptor superfamily, has been implicated in the regulation of immunity and inflammation in rodents and humans. The objective of this study was to investigate whether the increased expression of PPAR with fenofibrate treatment was altered in the immune system of mice brain tissue after Lipopolysaccharide (LPS) was injected intraperitoneally. Methods: PPARs expression was investigated in the brain tissues of BalB/c mice (n=6) under four situationssterile saline injection (control), LPS injection to generate periperal inflammation, LPS injection (5 mg/kg) with pretreated fenofibrate, and fenofibrate injection (100 mg/kg). Results: Fenofibrate has protective effects against the increase of LPS-induced proinflammatory cytokines including interleukin (IL)-1β, IL-6, and tumer necrosis factor-α in serum and brain tissue, measured using the enzyme linked immunosorbent assay, reverse transcriptase-polymerase chain reaction, and Western blot analysis. Also fenofibrate pretreatment markedly suppressed the reduction of PPARs mRNA and protein level by LPS injection. Moreover, the increased activation of PPARs abrogated the reduction of ABCD3, ACOX1, and catalase expression leading to an increase in the antioxidative capacity in brain tissue blocking the production of lipid peroxidation after LPS administration. Conclusion: The antiinflammatory and antioxidative properties of fenofibrate may be useful in ameliorating the progression of neuronal diseases. 연구배경: 다양한 신경계 이상 증상을 야기하는 Alzheimer’s disease, Parkinson’s disease와 같은 신경계 퇴행성 질환이나뇌졸중, 뇌 손상 및 자가 면역 질환들이 중추신경계의 염증반응과 연관이 있다는 논문이 발표되고 있다. 본 연구는 쥐의복강에 LPS를 주사함으로써 유발된 염증성 시토카인이 뇌조직에서 염증성 시토카인 발현을 증가시키고, 이 때문에전사 조절 인자인 PPARs 및 이들의 표적 유전자인 항산화효소의 발현이 감소하는지 확인하였으며, 이후 PPARs 활성화를 유발하는 fenofibrate가 중추신경계에서 이러한 현상을효과적으로 억제하는 지를 알아 보고자 하였다. 방법: Balb C 생쥐에 LPS (5mg/kg)와 fenofibrate를 단독 또는 병합 처리하여 4개군(LPS 단독 처리군, 대조군, fenofibrate 단독 처리군, LPS 병합 처리군)으로 구분하였다. 이후각 실험군에서 정해진 시간에 혈액과 뇌 조직을 채취하여RT-PCR, 웨스턴 블롯 및 면역조직 화학 염색법을 시행하였고,시토카인 및 PPARs의 발현 정도를 비교하였다. 결과: 혈액 및 뇌 조직 검사에서 fenofibrate가 LPS에 의한염증성 시토카인(IL-1β, IL-6 및 TNF-α)의 증가를 억제하는것으로 나타났다. 또한 fenofibrate는 뇌조직 검사에서 LPS에의한 PPARs의 발현 억제를 방지할 뿐만 아니라, 과산화소체와관련된 유전자(ABCD3, ACOX1 및 Catalase) 발현 감소도억제하는 효과가 있었고, oxidative stress 결과 산물인 3-NT와4-HNE의 발현을 감소시키는 효과도 나타내었다. 결론: 이번 실험을 통해 중추신경계 염증 반응을 유발할수 있는 알츠하이머병과 파킨슨병과 같은 만성 신경계 퇴행성질환 및 뇌졸중, 뇌 손상, 자가 면역질환 등에서 앞으로 fenofibrate의항염증 효과를 이용한 예방 및 치료제를 개발하는데도움이 될 수 있기를 기대한다.