• 대한한방부인과학회지
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• 제22권1호
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• pp.31-40
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• 2009

Purpose: This study was carried out to investigate the anti-tumor metastasis effect and activation of innate immunity by extracts of Mori radicis cortex. Methods: Anti-tumor metastatic experiment was conducted in vitro and in vivo by using colon 26-M3.1 carcinoma cell, L5178Y-R lymphoma cell and HeLa cell. To observe the activation of innate immunity by extracts of Mori radicis cortex, 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 Mori radicis cortex extracts significantly inhibited tumor metastasis. In an in vitro cytotoxicity analysis, Mori radicis cortex affected tumor cell growth above specific concentration. Mori radicis cortex also stimulated peritoneal macrophage, which was followed by the production of various cytokines such as IL-6, IL-10, IL-12, TNF-${\alpha}$. The depletion of NK cells by anti-asialo GM1 serum partly abolished the inhibitory effect of Mori radicis cortex on tumor metastasis. Conclusion: Mori radicis cortex appears to have considerable activity on the anti-metastasis by activation of innate immunity.

• Journal of Microbiology and Biotechnology
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• 제16권4호
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• pp.584-589
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• 2006

Bifidobacteria has been suggested to exert health promoting effects on the host by maintaining microbial flora and modulating immune functions in the human intestine. We assessed modulatory effects of the different cell fractions of Bifidobacterium sp. BGN4 on macrophage cells and other immune cells from the spleen and Peyer's patches (PP) of mouse. Cell free extracts (CFE) of the BGN4 fractions induced well-developed morphological changes in the macrophages and increased the phagocytic activity more effectively than other fractions in the mouse peritoneal cells. Nitric oxide (NO) production was significantly reduced by both the cell walls (CW) and CFE in the cultured cells from the spleen and PP. The production of interleukin-6 (IL-6) and interleukin-10 (IL-10) was eminent in the spleen cells treated with experimental BGN4 cell fractions. However, in the PP cells, IL-6 was slightly decreased by the treatment with the whole cell (WC) and CW, whereas IL-10 was significantly increased by the treatment with the CW and CFE. These results suggest that different types of bifidobacterial cell fractions may have differential immunomodulatory activities depending on their location within the host immune system.

• 대한한방내과학회지
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• 제19권2호
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• pp.208-218
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• 1998

To clarify the activating effects of Buthus martensi Karsch on immunological function, its effect on primary and secondary antibodies production in mice of various ages was investigated. Buthus martensi Karsch increased the number of both antibody producing cells(anti-IgM and anti-IgG producing plaque forming cells, PFC) and phagocytic activity of peritoneal macrophage. Futhermore, these phenomena were significantly increased with aging in mice. Buthus martensi Karsch also increased natural killer cell activity concerning to cancer immunology. These results suggest that Buthus martensi Karsch markedly increases the reduced activity in the elderly and activates the immune response in senescence mice.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.242-242
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• 1994

Cortex mori (Morus alba L.), the root bark of mulberry tree, has been used as an antiphlogistic, diuretic, and expectorant in herbal medicine. The purpose of this study was to determine whether Cortex mori could inhibit the ovalbumin (OA) -induced late asthmatic reaction in guinea pigs. Guinea pigs were sensitized by two exposures to an aerosol of OA(1.0%) and then challenged with aerosolized antigen(2.0%), The animals were pretreated by three inhalations of the aerosoled Cortex mori either before antigen sensitization or cahllenge. Bronchoalveolar lavage fluid(BALF) and peripheral blood were collected at 17 hours after OA challenge. The cell populations in BALF and peripheral blood were examined to determine the changes of the relative proportions of eosinophils,neutrophils and mononuclear cells etc. Beta-glucuronidase activity in BALF was measured to evaluate the alveolar macrophage activation. OA-induced histamine release from guinea pig peritoneal fluid cells was measured by radioisotope enzymatic asssay. Results were as follows. The number of eosinophils, neutriphils and lymphocytes recovered in BALF were significantly increased in the 17h following aerosol challenge with OA. Among them, eosinophil and neutriphils were decreased remarkably in group that had been preinhalated with Cortex mori. The number of lymphocytes in BALF were not decreased in group pretreated with CM before sensitization but decreased in Group pretreated with CM before challenge. After OA challenge, the number of eosinophils in peripheral blood were markedly increased, but Cortex mori inhibited significantly the OA-induced eosinophilia. Beta-glucuronidase activity in the supernatants of BALF were significantly increased in the 17h following aerosol challenge with OA, however, pretreatment of Cortex mori had no influence on Beta-glucuronidase activity, suggesting that Cortex mori had no inhibitory effect on OA-induced alveolar macrophage activation. Cortex mori inhibited the OA-induced histamine release from guinea pig peritoneal fluid cells. From the above results, it is suggested that Cortex mori contains some substances with an activity to inhibit the the OA-induced late phase reaction of the bronchial asthma in guinea pigs.

• 대한본초학회지
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• 제27권6호
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• pp.23-28
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• 2012

Objectives : Samhwangsashim-tang(SHSST), a mixture of Rhei radix et rhizoma, Scutellariae radix, and Coptidis rhizoma, has been regarded as being able to treat bleeding, gastric discomfort, dry mouth, insomnia and purpura due to Blood Heat. Currently, this herbal formula is applied to gastritis, gastric ulcer, hypertension, atherosclerosis or other types of vascular inflammatory disorders. Methods : We extracted this herbal mixture with 30% ethanol and examined for its effects on systemic inflammatory responses and in vitro macrophage activity. Mice were orally given to SHSST for 7 days and then lipopolysaccharide(LPS) was intraperitoneally injected. Tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) levels in serum were measured 1 h after LPS challenge. Peritoneal macrophages were isolated from thioglycollate-injected mice and used for in vitro cellular activity. Cell death was measured using the MTT method and annexin V/propidium iodide staining. LPS-stimulated signaling molecules necessary for TNF-${\alpha}$ expression were determined by Western blotting. Results : Oral administration of SHSST for 7 days resulted in a significant reduction in LPS-stimulated TNF-${\alpha}$ release into serum. In vitro treatment of SHSST was cytotoxic in a concentration-dependent manner. However, SHSST caused a concentration-dependent reduction in necrosis and increase in apoptosis in mouse peritoneal macrophages. SHSST inhibited the activation of NF-${\kappa}B$, p38 and JNK signaling molecules in response to LPS. Conclusion : Taken together, our results demonstrated that SHSST was effective in lowering LPS-stimulated TNF-${\alpha}$ serum levels, possibly through its modulation of NF-${\kappa}B$, p38 and JNK in macrophages.

• 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 .

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.350-356
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• 2017

We previously reported that the CopA3 peptide (LLCIALRKK, ${\small{D}}-form$) originally isolated from the Korean dung beetle has antimicrobial and immunosuppressive effects. However, the high cost of producing the synthetic peptide, especially the ${\small{D}}-form$, has limited the development of CopA3 for therapeutic purposes. Here, we investigated whether the CopA3 deletion derivative, CopA5, which is composed of only five amino acids (LLCIA) and has the ${\small{L}}-form$ structure, could inhibit the lipopolysaccharide (LPS)-induced activation of macrophages. Peritoneal exudate macrophages (PEM) were isolated from mice and exposed to LPS in the presence or absence of CopA5, and biomarkers of macrophage activation were measured. Our results revealed that LPS-induced nitric oxide (NO) production, tumor necrosis factor $(TNF)-{\alpha}$ secretion, and phagocytic activity of PEM were significantly inhibited by CopA5 treatment. Similar to CopA3, the structurally modified CopA5 peptide had no cell toxicity (as assessed by measurement of cell viability loss and apoptosis) in PEM. Moreover, the LPS-induced upregulation of the activating phosphorylation of signal transducer and activator of transcription 1 (STAT1) was markedly inhibited by CopA5 treatment. These results suggest that, similar to CopA3, CopA5 inhibits macrophage activation by inhibiting STAT1 phosphorylation and blocking the release of NO and $TNF-{\alpha}$. CopA5 may therefore prove therapeutically useful in the realm of immune suppression.

• 한국식품과학회지
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• 제48권3호
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• pp.289-295
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• 2016

국내 시판중인 인스턴트 커피에서 다당류를 추출하여 새로운 면역 활성 물질을 확인하고자 하였다. 시판중인 인스턴트 커피를 구입하여 냉수추출 후, 에탄올 침전과 투석을 통하여 약 24%의 높은 수율로 다당류(ICP-0)를 얻을 수 있었다. ICP-0은 중성당 89.9%, 산성당 10.1%로 이루어진 다당체로써, 구성 성분으로는 갈락토스 55.5%, 만노스 25.7%와 소량의 아라비노스, 람노스 등으로 이루어진 것을 확인하였으며, 갈락토만난(galactomannan)이나 팩틴 다당류(pectic polysaccharide)가 혼합된 구조일 것이라고 추정되었다. 인스턴트 커피 유래 ICP-0에 대하여 대식세포 자극에 대한 세포증식능을 확인한 결과, NC 대비 유의적으로 높은 증식활성을 보였으며, IL-6, IL-12 및 TNF-${\alpha}$의 경우, $10{\mu}g/mL$에서도 NC 대비 우수한 사이토카인 분비 유도능을 보임이 확인되었다. 또한 NO, ROS를 분석한 결과에서도 NO의 경우 $10{\mu}g/mL$ 이상에서 NC 대비 양호한 활성을 보여 주었다. ICP-0에 대하여 2차 면역기관 중 하나인 비장세포상의 증식능을 확인한 결과, NC 대비 115% 정도 증식하는 것이 확인되어 면역세포를 직접 증진시키는 마이토겐 활성이 있음을 나타내었다. 인스턴트 커피 유래 ICP-0가 장관 내 분포되어 있는 Peyer's patch를 경유하여 장관면역에 공헌할 수 있는지 여부를 확인한 결과, NC 대비 유의적으로 골수세포의 증식능을 확인하였으며 GM-CSF와 같은 관련 사이토카인의 분비 유도가 확인되었다. 이상의 결과는 시판 인스턴트 커피로부터 분리한 다당류가 대식세포의 기능과 장관면역계를 증진시킬 수 있는 가능성을 제시한다.

• 한국식품과학회지
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• 제46권1호
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• pp.68-72
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• 2014

본 연구는 동충하초(Cordyces militaris) 유래의 기능성 물질인 코디세핀의 면역활성을 검증하기 위하여 C57BL6 마우스 복강 대식세포를 이용하여 코디세핀이 대식세포의 활성화에 미치는 영향에 대하여 시험하였다. 그 결과 LPS에 의해 유도된 마우스 복강세포는 코디세핀의 작용에 의해 IL-$1{\beta}$, IL-12, TNF-${\alpha}$의 염증성 사이토카인의 생성이 증대되어 초기 염증매개 반응을 유도하여 선천면역반응의 활성화와 그리고 면역작용에 있어 후기 적응면역의 전환으로의 T 림프구의 활성화가 예상된다. 또한 IL-6의 생성증대로 활성화된 T 림프구에 의해 B 림프구의 항체생성반응을 매개하는 면역반응도 상승할 것으로 사료된다. 그리고 대식세포에 의한 염증반응에서 염증매개인자인 NO와 $H_2O_2$의 생성을 증대시킴에 따라 대식세포의 독성작용을 활성화시켜 염증반응을 효과적으로 유도할 것으로 보이며, 또한 $H_2O_2$의 후기 생성을 저해하였는데 이는 염증반응에 유도될 수 있는 세포의 손상으로부터 세포를 보호할 수 있을 것으로 사료된다. 따라서 코디세핀은 외부인자로부터 염증매개성 면역반응의 증강작용을 나타내는 것으로 사료된다.

• 한국군사과학기술학회지
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• 제16권3호
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• pp.390-397
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• 2013

Primary-cultured immune cells are widely used in research to elucidate the mechanism of inflammation including chemotaxis, production of reactive oxygen species, cytokine release and antigen presenting. Mice are one of the species of experimental animals commonly used for such studies. Immune cells can be isolated and cultured from various organs such as bone marrow, peritoneal cavity, lung, spleen. For elaborated experimental studies, immune cells should be elicited with inflammatory substances or proliferated in vitro with special media. This paper details methods of obtaining immune cells from various organs of mice and investigating immune mechanism using isolated immune cells. It contains standard protocols of isolating and culturing immune cells from bone marrow, peritoneal cavity and lymphoid organs. It also covers the methods of investigating immune mechanism such as ELISA, western blotting, confocal microscopy and ELISPOT assay. With the works in this study, we established the standardized isolation and analysis methods of primary-cultured immune cells.