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

• 동의생리병리학회지
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• 제36권1호
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• pp.1-6
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• 2022

The tuber of Pinellia ternata (Thunb.) Brei (TPT) used in traditional Oriental medicine for the treatment of cough, sputum, vomiting, and insomnia, possesses antioxidant, antibacterial, and anti-inflammatory effects. Although recent studies have reported the anticancer effects of TPT in several cancer cells, it is still unclear whether TPT regulates tumor-associated macrophage (TAM) characterized by the immunosuppressive M2 macrophage phenotype. Our results showed that the ethanol extract of TPT (ETPT) suppressed the migration of RAW264.7 mouse macrophage cells and THP-1 human monocytes differentiated into macrophages towards the conditioned media (CM) collected from lung cancer cells, suggesting that ETPT would attenuate the recruitment of macrophages into tumors. In addition, ETPT suppressed the interleukin (IL)-4 or IL-6-induced M2 macrophage polarization in RAW264.7 cells. ETPT treatment not only downregulated the mRNA expression of M2 macrophage markers including arginase-1, mannose receptor C type 1 (MRC-1), and IL-10, but also inhibited the phosphorylation of signal transducer and activator of transcription 3 (STAT3) and STAT6, general regulators of M2 macrophage polarization. Finally, the transwell assay results showed that the CM from M2-polarized RAW264.7 cells increased the migration of mouse lewis lung carcinoma (LLC) cells, while those from RAW264.7 cells co-treated with ETPT and IL-6 significantly reduced the migration of LLC cells. Taken together, our observations clearly demonstrate that ETPT suppressed the cancer cell migration by regulating macrophage recruitment and M2 macrophage polarization.

• 대한한방부인과학회지
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• 제22권2호
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• pp.79-93
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• 2009

Purpose: This research aimed to study the effect of FAE(Ferment Artemisiae Argyi Folium and Epimedii Herba) on the mouse macrophage cell activity. Methods: Effect of FAE, which was fermented by Sacchromyces cerevisiae STV89, on cell viability, amount of $H_2O_2$ within cells, amount of NO was measured and compaperd by using mouse macrophage cells. Results: 1. Result of MTT assay conducted to observe the effect of FAE on the survival rate of mouse macrophage cells illustrated that, when FAE was proccessed for each concentration, there was no significant decrease of the survival rate. 2. FAE increased the amount of $H_2O_2$ within macrophage cells and increased inhibition of amount of $H_2O_2$ in macrophage induced by LPS. 3. FAE inhibited amount of NO in macrophage cells, and significantly inhibited increase of amount of NO in mcacrophage induced by LPS. Conclusion: FAE produced by Artemisiae Argyi Folium and Epimedii Herba did not induce the decrease of macrophage cell survival rate, increased amount of $H_2O_2$ within cells, and reduced amount of NO. FAE significantly increase by LPS, reduced the increase of amount of NO in macrophage induced by LPS. These results signify FAE has significant effect on immuno modulating activity of macrophage.

• 대한약침학회지
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• 제22권4호
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• pp.253-259
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• 2019

Objectives: It is reported that tumor-associated macrophages (TAMs) contribute to cancer progression by promoting tumor growth and metastasis. The purpose of this study is to investigate the effect of different fractions of Adenophora triphylla var. japonica (AT) on the polarization of macrophages into the M2 phenotype, a major phenotype of TAMs. Methods: We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AT. The cytotoxicity of AT in RAW264.7 cells was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RAW264.7 cells were polarized into the M2 phenotype by treatment with interleukin (IL)-4 and IL-13. The expression of M2 macrophage marker genes was detected by reverse transcription polymerase chain reaction (RT-PCR). The phosphorylation level of signal transducer and activator of transcription 6 (STAT6) was investigated by western blot analysis. The migration of Lewis lung carcinoma (LLC) cells was examined by transwell migration assay using conditioned media (CM) collected from RAW264.7 cells as a chemoattractant. Results: Among various fractions of AT, the ethyl acetate fraction of AT (EAT) showed the most significant suppressive effect on the mRNA expression of M2 macrophage markers, including arginase-1, interleukin (IL)-10 and mannose receptor C type 1 (MRC-1), up-regulated by treatment of IL-4 and IL-13. In addition, EAT suppressed the phosphorylation of STAT6, a critical regulator of IL-4 and IL-13-induced M2 macrophage polarization. Finally, the increased migration of Lewis lung carcinoma (LLC) cells by CM from M2-polarized RAW264.7 cells was reduced by CM from RAW264.7 cells co-treated with EAT and M2 polarization inducers. Conclusion: We demonstrated that EAT attenuated cancer cell migration through suppression of macrophage polarization toward the M2 phenotype. Additional preclinical or clinical researches are needed to evaluate its regulatory effects on macrophage polarization and anti-cancer activities.

• 약학회지
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• 제44권2호
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• pp.175-181
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• 2000

Pedunculagin is an ellagitannin purified from Alnus hirsuta var. microphylla, Betulaceae. The effects of pedunculagin on the immune system have been characterized to induce enhancement of NK (natural killer) cell cytotoxicities against tumor cells. The present study investigated whether pedunculagin can enhance macrophage cytotoxicity against P8l5 tumor cells. Macrophage cultured with pedunculagin enhanced cytotoxicity in a dose dependent manner In addition, the same treatments increased NO production, which plays important roles in the immune system. liken together these results demonstrate that pedunculagin significantly enhances cytolytic activities of macrophage.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2022년도 추계학술대회
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• pp.88-88
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• 2022

In this study, we investigated the effect of Solanum nigrum aerial parts (SNAP) on macrophage activation and macrophage autophagy in RAW264.7 cells. SNAP increased the production of immunostimulatory factors and phagocytosis in RAW264.7 cells. TLR4 inhibition blocked SNAP-mediated production of immunostimulatory factors. In addition, the JNK inhibition reduced the SNAP-mediated production of immunostimulatory factors, and the SNAP-mediated JNK activation was blocked by the TLR4 inhibition. SNAP activated macrophage autophagy. TLR4 inhibition blocked SNAP-mediated macrophage autophagy and inhibition of p38 and JNK attenuated SNAP-mediated macrophage autophagy. These findings indicate that SNAP may induce TLR4/JNK-mediated macrophage activation and TLR4/p38 and JNK-mediated macrophage autophagy.

• 방사선산업학회지
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• 제4권3호
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• pp.227-231
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• 2010

The purpose of this study was to investigate the expression mechanism of MCP-1 in gamma-irradiated RAW 264.7 macrophage cells. MCP-1 plays an important role in attracting monocyte to injured site at the early inflammation stage. However the production mechanism of MCP-1 by gamma-irradiation in RAW 264.7 macrophage cells was almost undiscovered. We found that MCP-1 was produced in RAW 264.7 macrophage cells by irradiation with 5 Gy. And these inceases were attenuated by specific inhibitors treatment, such as $NF-{\kappa}B$, JNK, ERK, JAK2, and Pyk2. These results indicate that radiation-induced MCP-1 production is mediated by MyD88- and TRIF-dependent pathways in RAW 264.7 macrophage cells. Furthermore, gamma-irradiation induced heme oxygenase-1 (HO-1) expression in RAW 264.7 macrophage cells. However this induction level was reduced before MCP-1 and $IFN-{\beta}$ production.

• 대한본초학회지
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• 제23권2호
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• pp.151-157
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• 2008

Objectives : The purpose of this study is to investigate the biological Effect of multi-herbal drug 'KOCO-P1' on mouse macrophage Raw 264.7 cells. Methods : Multi-herbal drug 'KOCO-P1' was composed of Ginseng Radix, Astragali Radix, Polygonati Rhizoma, Liriopis Tuber, and Scrophulariae Radix. Cytotoxicity and cytoprotective activity of K0C0-P1 was verificated by MTT assay. And antioxidative effect of K0C0-P1 against EtOH, Nicotine was inspected by Hydroperoxide assay. Results : K0C0-P1 showed no cytotoxicity on RAW 264.7 cells for 24, 48, 72 hours. KOCO-P1 at 200, 100, and 50 ug/mL reduced the production of H202 in Raw 264.7 cells by EtOH. KOCO-P1 at 50 ug/mL reduced the production of H202 in Raw 264.7 cells by Nicotine. Conclusions : KOCO-P1 could be supposed to have antioxidative effect on macrophage with no cytotoxicity.

• 한국환경농학회지
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• 제28권3호
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• pp.295-300
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• 2009

포름알데히드는 농약 노출 시에 나타나는 중요한 물질로 천식 및 알러지등의 호흡 질환을 일으키는 물질로 알려져 있으며, 폐에서 대식세포는 면역 반응에 있어서 방어 기능을 담당하는 세포로 알려져 있다. 그러나 대식세포에서 포름알데히드에 대한 효과는 알려져 있지 않고 있어서 대식 세포주인 Raw 264.7 세포를 이용하여 실험하였다. 실험 결과 포름알데히드는 세포 생존율을 감소 시켰으며, 이러한 반응은 항산화제인 vitamin C, NAC, 및 catalase 처리 시 차단되었다. 실제로 포름알데히드 처리시 산화성 스트레스 지표인 lipid peroxide 형성이 증가하였으며 이들 반응 역시 항산화제들에 의해 차단되었다. 한편 포름알데히드 처리시 세포 사멸 촉진 단백질인 Bax 발현은 증가하였으며 세포 사멸 억제 단백질인 Bcl-2의 발현은 억제되었으며 이러한 반응은 항산화제 처리시 차단되었다. 세포사멸 실행 단백질인 casapse-3의 활성형 역시 증가하였으며, 항산화제 처리시 차단되었다. 결론적으로 포름알데히드는 폐 대식세포에서 산화성 스트레스 증가를 통해 세포 사멸을 일으키는 것으로 나타났다.

• 대한본초학회지
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• 제29권5호
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• pp.83-90
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• 2014

Objectives : This research has been done to investigate the anti-inflammatory effect of Coptidis Rhizoma extracts. Method : Coptidis Rhizoma was extracted by $100^{\circ}C$ water. The extract (CC : Extract of Coptis chinensis rhizome) was used to examine its effects on the cell viability of mouse macrophage Raw 264.7 cell line. Also the production of nitric oxide (NO), the c-jun N-terminalkinase (JNK) activation and the production of cytokines such as (IL)-5 were evaluated in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells. After the CC and LPS were applied to Raw 264.7 cells which were cultured for 24 hours, the MTT assay was performed. Result : The CC extracts didn't affect the viability of macrophage cells. However, the extracts inhibited the NO production and the JNK activation significantly in LPS-stimulated macrophage cells treated with 100 and $200{\mu}g/mL$ concentrations. The CC extract, also, impeded the production of inflammation-related factors and cytokines such as KC, VEGF, MCP-1, GM-CSF, IL-$1{\alpha}$, IL-5, IL-6, and IL-12p40 in LPS-stimulated macrophage cells at the concentration higher than $25{\mu}g/mL$. The production of basic-FGF concentration of 50 and $100{\mu}g/mL$, the production of IP-10 at $100{\mu}g/mL$, and the production of IFN-${\gamma}$ at $25{\mu}g/mL$, respectively. Conclusion : The CC prepared using $100^{\circ}C$ water showed the significant anti-inflammatory effect such as the inhibition not only on the production of NO, KC, VEGF, MCP-1, GM-CSF, IL-$1{\alpha}$, IL-5, IL-6, and IL-12p40 in LPS-stimulated macrophage cells at or higher than the concentration of $25{\mu}g/mL$, but also on the JNK activation at 100 and $200{\mu}g/mL$.