• Environmental Analysis Health and Toxicology
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• 제15권3호
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• pp.69-80
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• 2000

ADP-rubosylation may be involved in the process of macrophage activation. Nitric oxide (NO) has emerged as an important intracellular and interacellular regulatory molecule with function as diverse as vasodilation, neural communication or host defense. NO is derived from the oxidation of the terminal guanidino nitrogen atom of L-arginine by the NADPH -dependent enzyme, nitric oxide synthase (NOS) which is one of the three different isomers in mammalian tissues. Since NO can exert protective or regulatory functions in the cell at a low concentration while toxic effects at higher concentrations, its role may be tightly regulated in the cell. Therefore, this paper was focused on signal transduction pathway of NO synthesis, role of endogenous TGF-$\beta$ in NO production. effect of NO on superoxide formation. Costimulation of murine peritoneal macrophages with interferon-gamma (IFN-γ) and phorbol 12-myristate 13-acetate (PMA) increased both NO secretion and mRNA expression of inducible nitric oxide synthase (iNOS) when PMA abolished costimulation. Pretreatmnet of the cells with PMA abolished costimuation effects due to the depletion of protein kinase C (PKC) activities . The involvement of PKC in NO secretion could be further confirmed by PKC inhibitor, stauroprine, and phorbol ester derivative, phorbol 12,13-didecanoate. Addition of actinomycine D in IFN-γ plus PMA stimulated cells inhibited both NO secretion and mRNA expression of iNOS indication that PMA stabilizes mRNA of iNOS . Exogenous TGF-$\beta$ reduced NO secretion in IFN -γ stimulated murine macrophages. However addition of antisense oligodeoxynucleotide (ODN) to TGF-$\beta$ to this system recovered the ability of NO production and inhibited mRNA expression of TGF-$\beta$. ACAS interactive laser cytometry analysis showed that transportation of FITC -labeled antisense ODN complementary to TGF-$\beta$ mRNA could be observed within 5 min and reached maximal intensity in 30 min in the murine macrophage cells. NO released by activated macrophages inhibits superoxide formation in the same cells . This inhibition nay be related on NO-induced auto -adenosine diphosphate (ADP) -ribosylation . In addition, ADP-ribosylation may be involved in the process of macrophage activation .

• 동의생리병리학회지
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• 제22권1호
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• pp.171-175
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• 2008

In the recently, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Here we investigated the effect of the water extract of Loranthi Ramulus (LR) in production of inflammatory mediators and expression of toll-like receptor (TLR)-4, CD14 from peritoneal macrophage. We assayed the effect of LR water extract in cell proliferation in vitro and Th1/Th2 cytokine level in vivo. In peritoneal macrophages, water extract of LR water extract increased the production of Nitric oxide (NO) and $TNF-{\alpha}$. Also, LR water extract increased Con A-induced cell proliferation and IgG1, IgG2a level in serum. However, i.p. injection of water extract of LR water extract did not affect the level of $TNF-{\alpha}$, $IFN-{\gamma}$, IL-2, IL-4 and IL-5 in serum of mice. These studies indicate that LR water extract induces macrophage activation and suggest the possible use of LR water extract in macrophage-based immunotherapies.

• 한국식품영양과학회지
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• 제32권1호
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• pp.149-154
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• 2003

백작약은 한의학에서 보기ㆍ보혈을 위한 탕제에 이용되는 구성 생약재 중의 하나로서 본 연구에서는 백작약이 면역세포를 활성화시켜 암세포의 생장을 억제할 수 있는 능력이 있는지를 확인하고자 하였다. 그 결과 백작약 조다당분획(PJ-P)는 대식 세포를 활성화시켜 그 고유기능인 탐식 기능을 항진시켰다. 또한 암세포를 저해하는데 중요한 작용을 하는 NO와 TNF-$\alpha$, IL-1 그리고 IL-6의 분비를 향상시켰다. 이렇게 PJ-P에 의해 활성화된 대식 세포는 cytokine들과 NO를 생산함으로써 시험관 내에서 암세포를 살해하였으며, 또한 PJ-P는 암세포를 이식한 마우스의 생존기간을 연장시켰다 이같은 실험결과는 백작약의 조다당분획이 항암보조제 및 면역반응조절제로 활용될 가능성이 있음을 시사한다.

• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.376-395
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• 1996

The neuropeptide substance P(SP) has been recognized to modulate immune systems, with close proximity between peptidergic sensory nerve endings and immune cells. These include the macrophage and neutrophil activation, IL-2 production in T cell, augmentation of Ig synthesis, mast cell degranulation, $PGE_2$ and collagenase secretion in synoviocytes. In this study I examined SP-induced various biological activities such as antimicrobial action, cytokine production, and mast cell degranulation in the presence or absence of other inflammatory cell activators. Antimicrobial studies showed that undifferentiated HL-60 cells were not affected by SP. However, SP significantly enhanced antimicrobial action of TPA-treated or dbcAMP-treated HL-60 cells which had been differentiated into PMN or macrophage/monocyte. I could not find synergistic relationship between SP and LPS in parallel experiments of the above. SP did not induce IL-l production from murine macrophage cell line RAW264.7 whether costimulated with LPS or not. Mast cell degranulation was occured only when stimulated with high dose ($10^{-5}M$) of SP and the degree of this activation was slightly reduced by simultaneous application of $MIP-1{\alpha}$. In addition, CGRP which is known to be a common coexisting neuropeptide with SP within specific fibers did not augment the function of SP on mast cell degranulation. These results suggest that immunoregulatory activities of SP could be mediated through direct upregulation of various functions of immune cells and also upregulation of responsiveness of immune cells to other immune activators.

• Journal of Ginseng Research
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• 제43권1호
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• pp.154-160
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• 2019

Background: Compound K (CK) is an active metabolite of ginseng saponin, ginsenoside Rb1, that has been shown to have ameliorative properties in various diseases. However, its role in inflammation and the underlying mechanisms are poorly understood. In this report, the antiinflammatory role of CK was investigated in macrophage-like cells. Methods: The CK-mediated antiinflammatory mechanism was explored in RAW264.7 and HEK293 cells that were activated by lipopolysaccharide (LPS) or exhibited overexpression of known activation proteins. The mRNA levels of inflammatory genes and the activation levels of target proteins were identified by quantitative and semiquantitative reverse transcription polymerase chain reaction and Western blot analysis. Results: CK significantly inhibited the mRNA expression of inducible nitric oxide synthase and tumor necrosis factor-${\alpha}$ and morphological changes in LPS-activated RAW264.7 cells under noncytotoxic concentrations. CK downregulated the phosphorylation of AKT1, but not AKT2, in LPS-activated RAW264.7 cells. Similarly, CK reduced the AKT1 overexpression-induced expression of aldehyde oxidase 1, interleukin-$1{\beta}$, interferon-${\beta}$, and tumor necrosis factor-${\alpha}$ in a dose-dependent manner. Conclusion: Our results suggest that CK plays an antiinflammatory role during macrophage-mediated inflammatory actions by specifically targeting the AKT1-mediated signaling pathway.

• 대한한방부인과학회지
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• 제34권3호
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• pp.15-28
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• 2021

Objectives: This study was designed to examine the anti-cancer activity by innate immunomodulating and anti-inflammatory effects of liriopis tuber water extract (LPE). Methods: Cell cytotoxicity was tested with 4T1 mouse mammary carcinoma cells, spleen cells, macrophage, and RAW264.7 cells. To investigate innate immunomodulating effects of LPE on macrophage, we measured tumor necrosis factor-alpha (TNF-α), interleukin-12 (IL-12), and interleukin-10 (IL-10). To investigate innate immunomodulating effects of LPE on RAW264.7 cell, we measured TNF-α, interleukin-6 (IL-6). In addition, TNF-α and nitric oxide (NO) induced by lipopolysaccharide (LPS) were measured after treating with LPE to observe innate immunomodulating effect of LPE on RAW264.7 cell. Also, mitogen-activated protein kinase (MAPK) and nuclear factor κB (NF-κB) were examined by western blot analysis. Results: In an in vitro cytotoxicity analysis, LPE affected tumor cell growth above specific concentration. As compared with the control group, the production of TNF-α, IL-12, and IL-10 were increased in macrophage. As compared with the control group, the production of TNF-α and IL-6 were increased in RAW 264.7 cell. The expression of TNF-α and NO induced by LPS after treating LPE was decreased. In addition, treatment of RAW 264.7 cell with LPE increased the phosphorylation levels of p-extracellular signal-regulated kinase (p-ERK), p-Jun N-terminal kinase (p-JNK), and p-p38. Conclusions: LPE might have impact on the anti-cancer effect by activation of innate immune system and inflammation control.

• 대한한방부인과학회지
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• 제22권1호
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• pp.41-52
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• 2009

Purpose: This study was designed to investigate the anti-tumor metastasis by immunomodulating effects of extracts of Prunellae Spica. Methods: Antimetastatic experiment was conducted in vivo by using colon 26-M3.1 carcinoma. And we observed cytotoxicity of Prunellae Spica on colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell, hela cell and macrophage. To observe the immnomodulating effects of Prunellae Spica, we estimated IL-6, IL-10, IL-12, TNF-${\alpha}$ from peritoneal macrophages. And we evaluated the activation of NK cell by using anti-asialo-GM1 serum. Results: We found that the administration of Prunellae Spica extracts significantly inhibited tumor metastasis in vivo. In an in vitro cytotoxicity analysis, cell growth are closer to 100% in case of colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell, hela cell at low concentration. In case of macrophage, cell proliferation is closer to 100% less than $62.5{\mu}g/m{\ell}$ of Prunellae Spica extracts. The level of cytokine such as IL-6, IL-10, IL-12 which stimulates Prunellae Spica extracts was increased in dose-dependent manner compared to the control group. TNF-${\alpha}$ is hardly secreted less than $250{\mu}g/m{\ell}$ The depletion of NK cells by anti-asialo GM1 serum partly abolished the inhibitory effect of Prunellae Spica on tumor metastasis. Conclusion: Prunellae Spica appears to have considerable activity on the anti-metastasis by activation the immune system such as macrophage and NK cell.

• 대한약리학회지
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• 제29권1호
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• pp.107-120
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• 1993

입자 또는 용해성 자극 물질들은 칼슘 이동의 변화와 protein kinase C의 활성화를 초래하여 식 세포의 반응을 자극하는 것으로 추정하고 있다. 이에 비해서 protein kinase C가 활성화되면 호중구에서 agonist에 의한 세포 칼슘 농도의 증가가 억제된다고 보고하고 있다. PAF는 peritoneal macrophage에서 세포내 칼슘 농도를 용량에 따라 증가시켰으며 칼슘의 유출이 동반되었다. PAF에 의한 세포내 칼슘 농도의 증가는 TMB-8, verapamil과 TTX의 영향을 받지 않았다. TEA는 PAF에 의한 세포내 칼슘 이동을 자극하였으며 세포내 칼슘 농도의 감소를 지연시켰다. 5mM EGTA는 거의 완전히 PAF에 의한 세포내 칼슘 이동을 억제하였다. PAF의 첨가 후에 세포막 투과성은 반응 5분까지 현저하게 증가하였으며 이후 느리게 증가하였다. PAF에 의한 LDH 유리는 EGTA와 TMB-8에 의하여 약간 감소하였다. PAF에 의하여 자극된 superoxide 생성은 EGTA, TMB-8과 verapamil에 의하여 억제되었으나 TTX와 TEA의 영향은 받지 않았다. PAF에 의한 세포내 칼슘 농도의 증가, 세포막 투과성의 증가와 superoxide 생성은 IQSP, chlorpromazine과 propranolol에 의하여 억제되었다. PAF에 의한 LDH 유리는 chlorpromazine에 의하여 유의하게 그리고 propranolol에 의하여 다소 적게 억제되었다. PMA 전처리 후에 macrophage에서 세포내 칼슘 농도의 상승과 LDH 유리에 대한 PAF의 자극 효과는 유의하게 감소되었다. 이상의 결과로 부터 PAF는 세포내 칼슘 농도를 증가시키고 protein kinase C를 활성화시킴에 의하여 마우스 peritoneal macrophage에 자극 작용을 나타낼 것으로 시사된다. Protein kinase C를 미리 활성화시키면 macrophage 반응에 대한 PAF의 자극 작용은 억제될 것으로 추정된다.

• Archives of Pharmacal Research
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• 제25권6호
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• pp.910-916
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• 2002

Endotoxin tolerance reduces the capacity of monocytes to produce proinflammatory cytokines, results in cellular immune paralysis, and down-regulates the production of helper T (Th)1 type cytokines with a shift toward a Th2 cytokine response. Prostaglandin (PG)E$_2$ in the immune system also results in macrophage inactivation and the suppression of Th1 activation and the enhancement of Th2 activation. However, the inhibitory effects of PGE$_2$ on the altered polarization of the Th cell and macrophage interleukin (IL)-6 production characterized in part by cellular immune paralysis in a state of endotoxin tolerance is unclear. This study was undertaken, using indomethacin, to investigate the role of endogenous PGE$_2$ on the Th cytokines and macrophage IL-6 production in a state of endotoxin tolerance compared to those with endotoxemia mice, wherein, in this latter case, the increased production of proinflammatory cytokines and PGE$_2$ is exhibited. Endotoxemia was induced by injection of lipopolysaccharide (LPS; 10 mg/kg in saline) i.p. once in BALB/c mice, and endotoxin tolerance was induced by pretreatment with LPS (1 mg/kg in saline) injected i.p. daily for two consecutive days and then with LPS 10 mg/kg on day 4. Splenocytes or macrophages were obtained from endotoxemia and endotoxin tolerance models pretreated with indomethacin, and then cytokine production was induced by Con A-stimulated splenocytes for the Th cytokine assays and LPS-stimulated macrophages for the IL-6 assay. Our results showed that endotoxemia led to significantly reduced IL-2 and IL-4 production, to significantly increased IL-6 production, whereas interferon $(IFN)-{\gamma}$ production was not affected. Indomethacin in the case of endotoxemia markedly attenuated $IFN-{\gamma}$ and IL-6 production and didnt reverse IL-2 and IL-4 production. Endotoxin tolerance resulted in the significantly reduced production of IL-2 and $IFN-{\gamma}$ and the significantly increased production of IL-4 and IL-6. Indomethacin in endotoxin tolerance greatly augmented IL-2 production, significantly decreased IL-4 production, and slightly attenuated IL-6 production. These findings indicate that endogenous PGE$_2$ may mediate the suppressed Th1 type immune response, with a shift toward a Th2 cytokine response in a state of endotoxin tolerance, whereas endotoxemia may be regulated differentially. Also, endogenous PGE$_2$ may mediate macrophage IL-6 production in the case of endotoxemia to a greater extent than in the case of endotoxin tolerance.

• 생명과학회지
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• 제15권5호
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• pp.703-706
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• 2005

본 연구는 Wistar-Koyto Rat과 Spontaneously Hypertensive Rat을 대상으로 10주간의 L-arginine 투여와 규칙적인 수영 훈련을 하였을 때, 심박수, 혈압, 간조직의 eNOS 활성 및 대식세포의 활성수준을 비교 분석하여 고혈압 질환상태에서 심혈관계 및 eNOS 수준에 대한 L-arginine 투여와 훈련의 효과를 규명하는데 목적이 있었다. 결과적으로 고혈압 통제군의 심박수, 혈압, eNOS, 및 대식세포의 활성은 정산군에 비해 현저하게 높은 수치를 보였는데, 이는 고혈압으로 인한 혈관내피세포 손상으로 산화질소 생성기전에 불안정을 초래하여 심혈관계에 부정적인 영향을 주었을 것으로 해석한다. 하지만 L-arginine 투여와 규칙적인 수영훈련은 심박수, 혈압 및 eNOS 수준을 안정 수준으로 회복하였고, 대식세포의 활성 역시 안정화함으로 심혈관계에 긍정적인 영향력을 미치는 것으로 나타났다.