• 대한미생물학회지
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• 제35권3호
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• pp.251-261
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• 2000

V. vulnificus is an estuarine bacterium which causes septicemia and shock in susceptible patients. The organism produces a hemolytic cytolysin (VvH), which has a membrane damaging effect on erythrocytes. To clarify the mechanisms by which VvH might contribute to virulence, we examined its effect on macrophages. When mouse peritoneal macrophages were harvested and co-cultured with hemolysin-positive V. vulnificus strains (100 bacteria/cell), about 60% of the macrophages were killed; macrophages were not killed when co-cultured V. vulnificus strain CVD 707, a VvH-negative deletion mutant. Exposure of macrophages to filtered culture supernatants (2.5 HU/ml) and purified VvH (3 HU/ml) resulted in an increase in dead cells (80 and 90%, respectively), as determined by the trypan blue dye exclusion method and LDH release from macrophages was also increased (70 and 65.5%, respectively). The cytotoxic effect of VvH on macrophages was both the dose- and time-dependent. The VvH caused damage to the macrophage membrane and was blocked significantly by preincubation with cholesterol (p<0.01). Fetal bovine serum showed remarkable inhibition of VvH synthesis by V. vulnificus and inhibited VvH activity in culture supernatant. Cell viability was increased by 35% (p<0.01) and LDH release decreased by 28% (p<0.01) when macrophages were incubated with V. vulnificus (100 bacterial cell) in DMEM-10% FBS for 2 hr. Bacterial clearance activity of mice against V. vulnificus CVD 707 was decreased by pretreatment with 10 HU of VvH. This result suggests that the VvH can impair the membrane of macrophages and may playa role in the pathogenesis of V. vulnificus septicemia.

• Tuberculosis and Respiratory Diseases
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• 제47권4호
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• pp.451-459
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• 1999

배 경 : 패혈증에서 과도한 활성산소종의 생성은 급성 폐손상의 병리 기전에 중요한 역할을 한다. Catalase 및 MnSOD 등의 항산화 단백은 패혈증 환자의 혈청내 증가하며 급성호흡곤란증후군의 발생 예측 인자 및 패혈증의 예후 인자로 알려져 있다. Peroxiredoxin(Prx) 는 최근 독특하고 중요한 세포내 항산화 단백으로 알려져 있다. 본 연구는 대식세포인 mouse monocyte-macrophages(RAW 264.7) 세포에 산화 스트레스 및 내독소 처리후 Prx I 및 Prx II의 발현 평가하고 패혈증 동물 모델에서 복강세척액 및 기관지폐포세척액내 Prx I, Prx II 및 Trx의 양을 측정하였다. 방 법 : Prx I, Prx II 및 Trx에 대한 특이 항체를 이용하여 immunoblot 분석으로 호중구, 대식세포 및 적혈구에 이들의 분포를 평가하였다. Mouse monocyte-macrophages 세포에 $5\;{\mu}M$ menadione 및 1 mg/ml lipopolysaccharide(LPS) 을 처치하여 Prx I 및 Prx II의 발현을 평가하였으며 6 mg/Kg LPS를 복강내 투여하여 유발한 복강내 패혈증 동물의 복강세척액내 Prx I, Prx II 및 Trx의 양을 측정하였으며 복강내 패혈증 동물 및 5 mg/Kg LPS를 정맥내 투여하여 유발한 정맥내 패혈증 동물에서 기관지폐포세척액내 Prx I, Prx II 및 Trx을 측정하였으며 폐장의 염증 정도와 비교하였다. 결 과 : Prx I 및 Prx II의 분포는 호중구, 폐포대식세포 및 적혈구에서 서로 다른 양상을 보였다. Mouse monocyte-macrophages 세포에 $5\;{\mu}M$ menadione 및 $1\;{\mu}g/ml$ lipopolysaccharide을 처치하였을 때 Prx I 발현은 증가하였으나 Prx II 발현은 변화하지 않았다. 복강내 패혈증 동물에서 복강세척액내 Prx I, Prx II 및 Trx의 양은 증가하였으나 복강내 패혈증 및 정맥내 패혈증 동물에서 기관지폐포세척액내 폐장 염증 정도와 관계없이 Prx I, Prx II 및 Trx의 양은 증가하지 않았다. 결 론 : 세포내 항산화 단백으로서 mouse monocyte-macrophages 세포에서 Prx I의 발현은 산화 스트레스 및 내독소 처치후 증가한다. Prx I, Prx II 및 Trx양은 패혈증 동물 모델에서 국소 염증 부위에서 증가하나 손상된 폐장에서는 증가하지 않는다.

• 동의생리병리학회지
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• 제16권6호
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• pp.1284-1290
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• 2002

Scrophularia buergeriana Miquel (Scrophulariaceae) has been used as an anti-inflammatory drug in the folk medicine recipe and been proved its anti-inflammatory effect in the oriental medicine. Since nitric oxide (NO) and superoxide anion (O/sub 2//sup -/) are ones of the major inflammatory parameters, we studied the effect of aqueous extracts of Scrophularia buergeriana (SB) on NO and O/sub 2//sup -/ production in lipopolysaccharide (LPS)-stimulated mouse peritoneal macrophages. NO, O/sub 2//sup -/production and inducible NO synthase (iNOS) level were significantly reduced in LPS-activated macrophages by SB compared to those without. Electrophoretic mobility shift assay (EMSA) indicated that SB blocked the activation of NF-kB, which was considered to be a potential transcription factor for the iNOS expression. SB also blocked degradation of IkBα. Furthermore, IkB kinase alpha (IKKα), which phosphorylates serine residues of IkB directly, is inhibited by SB. These results suggest that SB could exert its anti-inflammatory actions by suppressing the activation of NF-kB through inhibition of IKK activity.

• Journal of Ginseng Research
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• 제42권1호
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• pp.68-74
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• 2018

Background: Ginsenosides have been reported to have many health benefits, including anti-inflammatory effects, and the resolution of inflammation is now considered to be an active process driven by M2-type macrophages. In order to determine whether ginsenosides modulate macrophage phenotypes to reduce inflammation, 11 ginsenosides were studied with respect to macrophage polarization and the resolution of inflammation. Methods: Mouse peritoneal macrophages were polarized into M1 or M2 phenotypes. Reverse transcription-polymerase chain reaction, Western blotting, and measurement of nitric oxide (NO) and prostaglandin $E_2$ levels were performed in vitro and in a zymosan-induced peritonitis C57BL/6 mouse model. Results: Ginsenoside $Rg_3$ was identified as a proresolving ginseng compound based on the induction of M2 macrophage polarization. Ginsenoside $Rg_3$ not only induced the expression of arginase-1 (a representative M2 marker gene), but also suppressed M1 marker genes, such as inducible NO synthase, and NO levels. The proresolving activity of ginsenoside $Rg_3$ was also observed in vivo in a zymosan-induced peritonitis model. Ginsenoside $Rg_3$ accelerated the resolution process when administered at peak inflammatory response into the peritoneal cavity. Conclusion: These results suggest that ginsenoside $Rg_3$ induces the M2 polarization of macrophages and accelerates the resolution of inflammation. This finding opens a new avenue in ginseng pharmacology.

• 한방안이비인후피부과학회지
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• 제17권1호
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• pp.163-171
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• 2004

The flowers of Lonicera japonica Thunb. (Caprifoliaceae) has been used as anti-inflammatory drug in the folk medicine recipe and been proved its anti-inflammatory effect in the oriental medicine. However, the action mechanism of Lonicera japonica that exhibits anti-inflammatory effects has not been determined. Since nitric oxide (NO) is one of the major inflammatory parameter, we studied the effect of aqueous extracts of Lonicera japonica (AELJ) on NO production in lipopolysaccharide (LPS)-stimulated mouse peritoneal macrophages. NO and inducible NO synthase (iNOS) level were significantly reduced in LPS-stimulated macrophages by AELJ compared to those without Electrophoretic mobility shift assay (EMSA) indicated that AELJ blocked the activation of nuclear factor kappa B (NF-kB), which was considered to be a potential transcription factor for the iNOS expression. AELJ also blocked the phosphorylation and degradation of inhibitor of kappa B-alpha (IkB-${\alpha}$). Furthermore, IkB kinase alpha (IKK${\alpha}$), which is known to phosphorylate serine residues of IkB directly, is inhibited by AELJ in vivo and in vitro. These results suggest that AELJ could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of NF-kB activity.

• 대한한의학회지
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• 제20권2호
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• pp.177-186
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• 1999

Okbyungpoongsan(OBPS) has long been known to have anti-allergic effect. In order to evaluate the influence on innate and specific immune response, the effects of OBPS on vascular permeability. hypersensitivities and phagocytic functions were measured. As the results, OBPS increased phagocytic activity of peritoneal macrophages in vitro and in vivo. But OBPS depressed formation of reactive oxygen intermediates(ROI) in vitro and in vivo, while the drug enhanced generation macrophages. Foot pad swelling in the mouse and contact hypersensitivity against dinitroflouorobenzene were decreased. OBPS had no effect on NK cells. But OBPS decreased vascular permeability induced by histamine without statistical significance. These results demonstrate that OBPS suppresses hypersensitivity reactions without affecting phagocytic functions and formation of ROI from macrophages. It also means that OBPS acts as a effective inducer to synthesis of nitric oxide which is effective for the infectious disease while it does damage to tissue less as it suppresses ROI, So we can conclude that OBPS could be used for the treatment of the disease related with immune function.

• Archives of Pharmacal Research
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• 제22권3호
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• pp.267-273
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• 1999

In this study we examined the potential for the synergistic augmentation of the antitumor activity of inflammatory mouse peritoneal macrophages by stimulation with protein A combined with $IFN-\gamma$. The moderate augmentative effect induced by preincubation with protein A was demonstrated to be concentration-dependent, whereas IFN-, had a very low activating effect. Following preincubation with both protein A and $IFN-\gamma$, a marked enhancement of macrophage activity was noted. In addition, based on the utilization of neutralizing antibody to TNF-$\alpha$ or the inhibition of NO Production, TNF-$\alpha$ and NO were proven to be involved as mediators during the activation of tumoricidal macrophages by protein A in combination with $IFN-\gamma$. We also demonstrated that supernatants from macrophages treated with protein A plus $IFN-\gamma$ contained both TNF-$\alpha$ and NO at markedly increased levels. Thus, tumor cell lysis in the combined system was mediated via TNF-$\alpha$ or NO. These results demonstrate the synergistic effects on mouse pertioneal macrophage function of protein A in combination with $IFN-\gamma$ and suggest that combinations of such agents may serve as the basis for future in vivo immunotherapy.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7289-7294
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• 2013

We here document discovery of a new and simple model of tumor seeding involving the mouse peritoneum. Irradiated tumor cells administered by i.p. injection provided effective vaccination against peritoneal carcinomatosis and distal metastasis with colorectal carcinomas. In flow cytometric analysis, CD4+ and CD8+ T lymphocytes, macrophages and myeloid-derived suppressor cells (MDSCs), which are easy to obtain in the peritoneal cavity, were revealed to have significant differences between immunized and non-immunized mice and these contributed to antitumor responses. We also observed that both serum and peritoneal lavage fluid harvested from immunized mice showed the presence of CT26-specific autoantibodies. In addition, increase in level of TGF-${\beta}1$ and IL-10 in serum but a decrease of TGF-${\beta}1$ in peritoneum was found. Taken together, these findings may provide a new vaccine strategy for the prevention of peritoneal and even systemic metastasis of carcinomas through induction of an autoimmune response in the peritoneum.

• BMB Reports
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• 제40권6호
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• pp.928-935
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• 2007

Three Angelica sinensis polysaccharide fractions (APFs), named APF1, APF2 and APF3, were isolated and purified from Radix A. sinensis and their antioxidant activities were evaluated in isolated mouse peritoneal macrophages by pretreatment with APFs before exposure to 0.2 mM tertbutylhydroperoxide (t-BHP). The results showed that pretreatment of the macrophages with APFs as low as $10{\mu}g$/ml could significantly enhance t-BHP-decreased cell survival, intracellular glutathione (GSH) content and superoxide dismutase (SOD) activity, and also inhibited t-BHP-increased lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) formation (p < 0.05), and APF3 was the most active fraction, followed by APF2 and APF1 in decreasing order. Furthermore, we found for the first time that the bound-protein in APF3 was associated closely with the protective effects and the polysaccharide inhibited the excess NO release from t-BHP-activated macrophages to protect host cells.

• 한국식품영양학회지
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• 제25권4호
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• pp.755-761
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• 2012

최근 산삼근의 배양이 실험실에서 가능해짐에 따라 산삼에 대한 많은 연구가 이루어지고 있다. 본 연구에서는 산삼 배양근으로부터 다당을 분리하여 보체계 및 macrophage에 대한 면역자극활성을 측정하였다. 산삼 배양근을 열수 추출한 뒤, 80% ethanol 침전을 통하여 조다당을 추출하여 특성을 검토한 결과, 산삼 배양근 조다당체(WGAR-CP)의 neutral sugar 함량은 64.77%였고, uronic acid 함량은 34.32%이었다. WGAR-CP의 구성당 조성을 확인한 결과, glucose를 높은 비율로 함유하고 있었으며, rhamnose, arabinose, galactose, glucose를 미량 함유하고 있었다. Uronic acid에서는 galacturonic acid를 높은 비율로 함유하고 있었다. 한편, WGAR-CP는 인체 초기 면역반응에 있어 중요한 역할을 담당하는 보체계에 대하여 농도 의존적인 활성을 나타냈다. WGAR-CP를 0.1, 1, 10, 50, $100{\mu}g/m{\ell}$ 농도로 mouse peritoneal macrophage에 처리하여 면역자극을 유도한 후, NO 생성 및 cytokine 분비활성을 평가하였다. 그 결과 WGAR-CP의 NO생성은 저농도에서는 유의적인 활성을 보이지 않았으나, 50, $100{\mu}g/m{\ell}$에서 농도별로 유의적인 차이를 보였다. 또한 50, $100{\mu}g/m{\ell}$ 용량에서 IL-6 생성을 농도 유의적으로 증가시켰고, 10, $50{\mu}g/m{\ell}$에서 농도 유의적으로 IL-12의 생성을 증가시켰다. 이상의 결과를 종합해 볼 때, WGAR-CP는 자연면역계를 구성하고 있는 보체계와 macrophage의 활성인자로 작용할 수 있음이 확인되었고, WGAR-CP에 의해 활성화된 macrophage는 NO 및 IL-6, IL-12와 같은 염증성 cytokine의 분비를 유도함으로써 체내 면역기능을 증강시킬 수 있는 가능성이 있을 것으로 사료되었다.