• Fisheries and Aquatic Sciences
• /
• 제6권3호
• /
• pp.130-134
• /
• 2003

Proinflammatory effects of bacterial lipopolysaccharide (LPS) have been assessed by analysing the induction of two inflammatory genes, $interleukin-1\beta$ $(IL-1\beta)$ and cyclooxygenase-2 (COX-2), in rainbow trout (Oncorhynchus mykiss) macrophage cells. Production of a metabolite of arachidonic acid by COX-2, prostaglandin $E_2\;(PGE_2)$, was also analysed in macrophage cells after LPS stimulation. Northern blot analysis revealed that LPS $(5{\mu}g/mL)$ significantly upregulated $IL-1\beta$ (54 times) and COX-2 (40.7 times) gene expression in macrophage cells after 4 h stimulation. According to RT-PCR (Reverse Transcription Polymerase Chain Reaction) analysis, $IL-1\beta$ gene induction in LPS stimulated macrophage cells was started within 1h and significantly increased thereafter until 4h. Meanwhile, COX-2 gene induction by LPS was delayed in comparison with $IL-1\beta$ gene induction as a faint band was observed after 4h stimulation in head kidney macrophage cells. LPS also significantly increased $PGE_2$ production in head kidney leucocytes, presumably via activating COX-2 expression that metabolites arachidonic acid to $PGE_2$. In conclusion, it was demonstrated that LPS could induce two main inflammatory and immune related genes, $IL-1\beta$ and COX-2, and increase $PGE_2$ production in trout head kidney macrophage cells, representing a strong inflammatory activity.

• The Korean Journal of Physiology and Pharmacology
• /
• 제10권3호
• /
• pp.161-165
• /
• 2006

NO and cyclooxygenase-2 (COX-2) are contributes to vascular inflammation induced by various stimulation. The mechanism, which explains a linkage between NO and COX-2, could be of importance in promoting pathophysiological conditions of vessel. We investigated the effects of NO donors on the COX-l and COX-2 mRNA/protein expression, as well as the nitrite production in culture medium of vascular smooth muscle cell (VSMC). VSMC was primarily cultured from thoracic aorta of rat. In this experiments, COX-l and COX-2 mRNA/protein expressions were analysed and nitrite productions were investigated using Griess reagent. VSMC did not express COX-2 protein in basal condition (Nonlipopolysaccharide (LPS) stimulated). In LPS-stimulated experiments, after 3 hours of NO donor pretreatment, LPS $10{\mu}g/ml$ was treated for 24 hours. COX-l protein expressions were unchanged by SNP and NOR-3. NOR-3 significantly increased COX-2 mRNA/protein expression under LPS stimulation. In contrast, SNP did not increase COX-2 mRNA/protein expression under LPS stimulation. Nitrite production was higher in NOR-3 treatment than SNP treatment under LPS stimulation. These results suggest that the expression of COX-2 in VSMC is regulated by NOR-3, COX-2 expressions were depending on the types of NO donor and LPS stimulation in VSMC.

• Journal of Microbiology
• /
• 제39권4호
• /
• pp.314-320
• /
• 2001

Macrophages stimulated by lipopolysaccharide(LPS) from gram negative bacteria undergo activation of a group of immediate early genes including Rantes. The mouse Rantes gene promoter region contains an LPS rsponsive element(LPE) We detected 3 specific bands termed B1, B2 and 3 formed by the interaction of the LPE and proteins found in LPS-stimulated RAW 367.7 cells. An additional band B4 was determined to be an Ap-1 binding protein. The B1 band appears within 1 hour of LPS nuclear extracts from LPS-stimulation, and this protein kinase enhances B1 and formation. The B1 band can be converted to band B2/B3 by adding specific heparin column fraction purified Ser/Thr specific protein phosphatases PP-1 and PP-2A can stimulate the same conversion to about the same extent. Thus, the formation of the LRE sequence binding complex appears to be regulated by Ser/Thr protein kinase and one or more Ser/Thr specific phosphatases. At least four proteins are involved in the trgulation of the LRE-dependent Rants experssion: two binding factors that bind directly to the target sequences. and two factors that control their binding. The future purification and characterization of these binding pro-teins will reveal in detail the mechanism of Rantes gene activation after LPS stimulation.

• Journal of Microbiology and Biotechnology
• /
• 제28권3호
• /
• pp.491-497
• /
• 2018

Lipopolysaccharide (LPS), a component of the cell wall of gram-negative bacteria, elicits the secretion of cytokines, such as interferons, that stimulate the host defense system. Previously, we demonstrated that interferons induce interferon regulatory factors (IRFs) 1, 3, and 7, which regulate the transcription of Noxa and alter the expression profiles of Bcl-2 family proteins in tumors. However, the immediate consequences of LPS stimulation on Noxa and BH3 expression in tumor cells remain uncharacterized. In this study, we determined that LPS induced Noxa expression in CT26 cells. Furthermore, studies in HCT116 parental and HCT116 p53-deficient cells revealed that LPS-mediated Noxa was independent of p53. Meanwhile, IRF1, 3, and 7 in CT26, HCT116 parental, and HT116 p53-deficient cells were upregulated by LPS stimulation, suggesting that LPS induces the expression of these IRFs in a p53-independent manner. The responsiveness of IRF1, 3, 4, and 7 binding to the Noxa promoter region to LPS indicated that IRF1, 3, and 7 activated Noxa expression, whereas IRF4 repressed Noxa expression. Together, these results suggest that LPS directly affects Noxa expression in tumor cells through IRFs, implicating that it may contribute to LPS-induced tumor regression.

• Asian-Australasian Journal of Animal Sciences
• /
• 제19권5호
• /
• pp.713-719
• /
• 2006

The effect of dietary iodine and selenium supplementation, alone or in combination, on peripheral blood lymphocyte function was determined in laying hens. Eight-hundred-and-sixty-four New-Loman laying hens were randomly allotted into 12 dietary treatments with different inclusion levels of iodine (0, 0.1 and 0.2 mg/kg), selenium (0, 0.05, 0.1 and 0.2 mg/kg) or their combinations for 24 weeks. The lipopolysaccharide (LPS) stimulation index, concanavalin A (ConA) stimulation index, peroxide enzyme activity and phagocytosis to neutral red particles were tested. There were significant differences in LPS stimulation index, ConA stimulation index, peroxide enzyme activity and phagocytosis to neutral red particles in different iodine or selenium supplementation levels (p<0.05). The highest iodine and selenium supplementation both resulted in highest LPS-/ConA-stimulation indices (p<0.05). However, when iodine was lower than 0.2 mg/kg, the additional effect of different levels of selenium did not always result in significant differences in these indices. The results indicated that iodine and selenium may affect immunity in laying hens and, when the iodine level in the laying hen is lower than 0.2 mg/kg, a selenium allowance higher than 0.1 mg/kg may be necessary to improve immunity.

• Toxicological Research
• /
• 제18권4호
• /
• pp.363-367
• /
• 2002

Reactive of gen species (ROS) contribute to several cellular function and are involved in the regulation of signal transduction, gene expression, and proliferation. In the present study, we investigated the effect of $H_2O_2$ treatment on IgM secretion in LPS-stimulated murine B Iymphoma, CH12.LX. Cells were treated directly With $H_2O_2$ and stimulated with LPS. $H_2O_2$ treatment during 72 h time period inhibited IgM secretion in LPS-stimulated CH12.LX cells in a dose- and time-dependent manners. After treatment with 50 $\mu\textrm{M}$ $H_2O_2$ during 72 h time period, the level of IgM in LPS-stimulated CH12.LX cells was markedly decreased, whereas cell viability was not significantly changed. Addition of $H_2O_2$ concomitantly with LPS, or 12 h post-LPS stimulation, produced a significant inhibition of IgM secretion, Whereas inhibitory effect of $H_2O_2$ on IgM secretion was not observed when added 24 h after LPS stimulation. These findings suggest that $H_2O_2$ can inhibit the secretion of IgM in LPS-stimulated CH15.LX cells, and may alter the events necessary for terminal B cell differentiation.

• Applied Microscopy
• /
• 제43권1호
• /
• pp.27-33
• /
• 2013

Inflammatory cells express the inflammatory cytokines and growth factors induced by lipopolysaccharide (LPS). Odontoblasts are located at the pulp-dentin interface and extend their cell processes far into the dentin where they are the first cells to encounter microorganisms or their products. Therefore, this study examined the expression of some growth factors related to the signal pathway, such as growth factor receptor binding protein 2 (Grb2)-Ras in odontoblast-like dental pulp cells, after a treatment with LPS. After 60 minutes, the mRNA and protein expression levels of Grb2 and Ras were higher in the LPS-treated cells than in the control cells. The level of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA expression was increased significantly to a level similar to that of Grb2 and Ras at 60 minutes. The platelet-derived growth factor-AA (PDGF-AA) mRNA level was expressed strongly in the odontoblast like dental pulp cells without an association with LPS stimulation. Scanning electron microscopy revealed many extensions of the cytoplasmic processes and the number of processes increased gradually at 30, 60 and 90 minutes after LPS stimulation. From these results VEGF and bFGF expression might be induced through the Grb2-Ras signal transduction pathway in LPS treated odontoblasts.

• Tuberculosis and Respiratory Diseases
• /
• 제62권4호
• /
• pp.299-307
• /
• 2007

배경: HMGB1은 염증반응의 후기에 분비되는 중요한 염증유발물질 중 하나이다. 본 연구에서는 기존에 염증유발물질로 잘 알려진 LPS와 새롭게 염증유발물질로 관심을 받고 있는 HMGB1의 염증유발작용을 생체 외 및 생체 내 실험을 통해 비교하고자 하였다. 또한 HMGB1의 자극에 의한 IL-8 promoter region의 활성화에 중요한 역할을 수행하는 전사인자들을 확인하고자 하였다. 방법: RAW264.7 세포에 LPS(100 ng/ml) 또는 HMGB1(500 ng/ml)을 투여하고 각각 0, 2, 4, 8, 12 그리고 24시간 뒤에 세포상층액의 $TNF-{\alpha}$, MIP-2 그리고 $IL-1{\beta}$의 농도를 ELISA법으로 측정하였다. 생쥐의 복강에 LPS(5 mg/kg) 또는 HMGB1(2.5 mg/kg)을 주입하여 급성폐손상을 유발한 후에 폐의 사이토카인의 발현과 MPO 활성도를 측정하였다(LPS는 4시간 뒤, HMGB1은 24 시간 뒤). IL-8 promoter 부위에 있는 NF-IL6, $NF-{\kappa}B$ 그리고 AP-1에 대한 결합부위에 대해 돌연변이를 일으킨 후에 각각의 돌연변이체를 pIL-6luc에 결합시킨 뒤 RAW264.7 세포에 삽입하였다. 이 세포들을 36시간 배양한 후에 HMGB1(500 ng/ml)으로 자극하고, 한 시간 뒤에 세포를 녹인 후 luciferase 활성도를 측정하였다. 결과: LPS 투여 후에 RAW264.7 세포 배양상층액의 $TNF-{\alpha}$농도는 24시간 뒤에, MIP-2 농도는 8시간 뒤에 최고치를 보였다. 한편 HMGB1 투여 후에는 $TNF-{\alpha}$와 MIP-2 농도 모두 24시간 뒤에 최고치를 나타내었다. LPS 복강 내 투여 후 4시간 뒤에 생쥐의 폐의 $TNF-{\alpha}$, MIP-2 그리고 $IL-1{\beta}$의 농도는 대조군에 비해 현저히 증가하였으나, HMGB1 복강 내 투여 후 24시간 뒤에 생쥐의 폐에서는 $IL-1{\beta}$의 농도만 약간 증가하였다. MPO 활성도는 LPS와 HMGB1 투여 후에 모두 증가하였으며, LPS 투여 후가 더 의미있게 증가하였다. $NF-{\kappa}B$ 돌연변이체와 AP-1 돌연변이체에서 luciferase 활성도가 의미있게 감소하였다. 결론: 이상의 결과를 살펴볼 때 HMGB1은 염증유발효과는 LPS에 비해 강도가 떨어지나 지속시간은 오래 계속되는 것으로 보이며, HMGB1에 의한 IL-8의 활성화에 $NF-{\kappa}B$ 뿐만 아니라 AP-1도 중요한 역할을 수행하는 것으로 판단된다.

• Journal of Acupuncture Research
• /
• 제18권1호
• /
• pp.100-112
• /
• 2001

Objective : The purpose of this study is to investigate the immunological effect of Ramulus Cinnamomum aqua-acupuncture on the cellular immune response in mice with LPS induced arthritis. Methods : All the BALB/C mice used in this study were bred and maintaned in our pathogen-free mouse colony and were 6wk of age at the start of the experiment. The experimental model of arthritis was induced by injeciton of 300${\mu}g$/kg LPS in mice knee joint. Ramulus Cinnamomum aqua-acupuncture was injected into Yangnungchon(Gb34) of mice 2daily for 14days. Immunohistochemical analysis was carried out to assess CD4+, CD8+, CD11b, IL-$1{\beta}$, IL-2R and CD106 expression in common iliac lymph nodes and synovial menbrane after stimulation with Ramulus Cinnamomum. Electron microscopy was carried out to assess change of synovial membrane. Ramulus Cinnamomum aqua-acupuncture stimulation group was compared to control group and non stimutated with aqua-acupuncture. Resutts : At day 14 post arthritis onset, Immunohistological studies using monoclonal antibodies showed that Ramulus Cinnamomum aqua-acupuncture goup had decreased expression of CD4+, CD8+, CD11b, IL-$1{\beta}$, IL-2R and CD106 at common illiac lymph nodes and synovial membrane compared with control group. Conclusions : Ramulus Cinnamomum aqua-acupuncture stimulation inhibited the development of cellular immunity to LPS-induced arthritis in mice. Thus, aqua-acupuncture stimulation may have preventive effects on autoimmune inflammatory joint diseases. The effects of AA on immune function and disease activity in patients with RA warrant further investigation.

• 미생물학회지
• /
• 제27권2호
• /
• pp.147-154
• /
• 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.