• Journal of Physiology & Pathology in Korean Medicine
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• v.38 no.1
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• pp.32-37
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• 2024

The aim of this study was to investigate the role of Daesiho-tang (Da Chai Hu-Tang) water extract (DSTE) in regulating chronic stress-induced cancer progression, focusing on its activity in modulating tumor-associated macrophages (TAMs). Different stimuli can polarize TAMs into immune-stimulating M1 macrophages or immunosuppressive M2 macrophages. During cancer progression, M2 phenotype increases and supports tumor growth, angiogenesis and metastasis. Notably, chronic stress-induced catecholamines promote M2 macrophage polarization. In this study, we investigated whether DSTE regulates norepinephrine (NE)-induced M2 macrophage polarization in RAW 264.7 mouse macrophage cells. Even though NE itself did not increase the expression of M2 markers, the conditioned media of NE-treated 4T1 mouse breast cancer cells (NE CM) significantly up-regulated M2 markers in RAW 264.7 cells, suggesting that NE-regulated cancer cell secretome stimulated M2 polarization. However, such increase was abrogated by DSTE. NE CM also induced phosphorylation of signal transducer and activator of transcription 6 (STAT6) in RAW 264.7 cells, which was clearly reversed by pretreatment with DSTE, demonstrating that DSTE inhibited M2 polarization by inactivating STAT6. Finally, M2-polarized RAW264.7 cells by NE CM markedly increased the migration of 4T1 cells. However, such increase was completely reversed by co-treating RAW264.7 cells with NE CM and DSTE, indicating that DSTE attenuated cancer cell migration by blocking M2 polarization. Taken together, our results suggest a probable use of DSTE for cancer patients under chronic stress by regulating M2 macrophage polarization.

• The Korea Journal of Herbology
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• v.21 no.2
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• pp.165-173
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• 2006

Objectives : Coptidis Rhizoma has been known traditional medicine with antimicrobial activities. We investigated inhibitory effects of Coptidis Rhizoma extract on lipopolysaccharide(LPS)-induced nitric oxide production from mouse macrophages. Methods : After Coptidis Rhizoma extract was pretreated in BV2, mouse brain macrophages and RAW264.7 mouse macrophages, cells were activated with LPS. To investigate cytotoxicity Coptidis Rhizoma extract, cell viability was measured by MTT assay. The production of nitric oxide(NO) and inducible nitric oxide synthase(iNOS) was determined in each culture supernatant and mRNA by Griess reaction and RT-PCR. The production of $TNF-{\alpha}$ from cells was measured by ELISA. Results : Coptidis Rhizoma extract significantly inhibited LPS-induced NO production in BV2 and RAW264.7 cells. Coptidis Rhizoma extract also greatly suppressed mRNA expression of iNOS in BV2 and RAW264.7 cells activated by LPS. Conclusion : These data suggests that Coptidis Rhizoma extract may have an anti-inflammatory effect through the inhibition of NO production.

• Journal of Digital Convergence
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• v.16 no.11
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• pp.613-620
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• 2018

This study was conducted to investigate of Mori Folium Water Extract (MF) on anti-inflammation activity. MF Water extracts after 24 houres cultivation were examined to ascertain the cell viability of mouse macrophage RAW 264.7 cells. The influence of the Water extracts in RAW 264.7 macrophage cells treated with LPS was investigated. nitric oxide (NO) production, nterleukin$(IL)-1{\alpha}$ IL-6 and IL-10 increased generation of cytokines. mouse macrophage RAW 264.7 cells cell viability changes were no decreas after MTT assay of MF Water extract. The MF water extracts inhibited NO generation caused by LPS in the macrophages over $25{\mu}g/mL$. The MF water extracts increased in the control group the $IL-1{\alpha}$ and IL-6 activation generated by LPS in the macrophages over $50{\mu}g/mL$. Accordingly, it was found that different MF water extract concentrations significantly influenced certain anti-inflammation activities in RAW 264.7 macrophage cells. The results of this study are expected to be highly applicable to health - friendly functional materials. Further studies are needed to confirm the signaling pathways associated with anti-inflammation of macrophages through continuous studies.

• The Korea Journal of Herbology
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• v.37 no.5
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• pp.53-61
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• 2022

Objectives : The aim of this study was to investigate the effect of Baicalein (BA) on the production of hydrogen peroxide in lipoteichoic acid-stimulated RAW 264.7 mouse macrophages. Methods : Lipoteichoic acid-stressed RAW 264.7 mouse macrophages were incubated with baicalein at concentrations of 50 and 100 µM. Incubation time is 30 minutes, 2 h, 12 h, and 18 h. After incubation, The production of hydrogen peroxide in RAW 264.7 mouse macrophages was measured with dihydrorhodamine 123 assay. Streptococcus aureus lipoteichoic acid and Streptococcus pyogenes lipoteichoic acid were used as cell-stimulating lipoteichoic acid. Cell viabilities were measured with a modified MTT assay. Berberine, indomethacin, and gallic acid were incubated for the same time as the comparative materials. Results : BA at the concentration of 50 and 100 µM did not show cytotoxicity on RAW 264.7 mouse macrophages for 24 h incubation. For 30 minutes, 2 h, 12 h, and 18 h incubation, BA at the concentration of 50 and 100 µM significantly inhibited the production of hydrogen peroxide in RAW 264.7 mouse macrophages stimulated by Streptococcus aureus lipoteichoic acid (p < 0.05); also, BA at the concentration of 50 and 100 µM also inhibited the productions of hydrogen peroxide in RAW 264.7 mouse macrophages stimulated by Streptococcus pyogenes lipoteichoic acid significantly (p < 0.05). Conclusions : BA might have anti-bacterial activity related to its inhibition of hydrogen peroxide production in lipoteichoic acid-stimulated RAW 264.7 mouse macrophages.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.5-14
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• 2003

Objective: Present study was aimed to verify the effect of granulocyte macrophage-colony stimulating factor (GM-CSF) in the preimplantation development of mouse embryos and the involvement of the mitogen activated protein kiase (MAPK) in the GM-CSF signaling. Methods: Two-cell embryos were cultured for 96 h in the presence or absence of GM-CSF (0, 0.4, 2, 10 ng/ml) and PD98059, a MEK inhibitor (10 ${\mu}M$). Morphological development, cell number per blastocyst, and apoptotic nuclei, were eamined. MAPK activity of embryonic immunoprecipitate by MAPK (ERK1/2) antibody was measured by in vitro phosphorylation of myelin basic protein. Results: At post hCG 122 h the embryonic development among the experimental groups was significantly different (p=0.018). The rate of blastocyst development and cell number per embryo were the highest in 2 ng/ml GM-CSF treatment group. The percent of apoptotic cells of the GM-CSF-treated embryos was the lowest among the group. In blastocysts, GM-CSF treatment transiently increased MAPK activity. PD098059 attenuated the effect of GM-CSF on the morphological development, increase in cell number per blastocyst, down regulation of apoptosis, and upregulation of MAPK activity, suggesting that activation of MAPK cascade possibly mediated the embryotropic effect of GM-CSF. Conclusion: This result suggested that GM-CSF potentiated the development of preimplantation mouse embryos by activation of MAPK.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.20 no.1
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• pp.25-36
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• 1994

It is weal known that bacterial lipopolysaccharide (LPS) stimulates prostaglandin synthesis in various experimental system via enhancing the expression of cylooxygenase-2 (COX-2). This study was designed to characterize U)5-induced prostaglandin synthesis in mouse peritoneal macrophages LPS-stimulated prostaglandin synthesis in macrophages with short term exposure was not so much prominent, but there was a burst in prostaglandin synthesis 8 hours after the LPS treatment and this u·as accompanied with the increase of cyclooxygenase activity, Dexamethasone markedly inhibited prostaglandin synthesis in this system. Metabolic label ins data supported above observations and thus, it could be concluded that LPS induces the do novo synthesis of COX-2 by which it stimulates the prostaglandin synthesis in mouse peritoneal macrophages, These data suggested that this experimental model system could be used for the screening procedure of COX-2 selective inhibitors. Ketoprofen, a non steroidal anti inflammatory agent, appeared to inhibit COX-1 relatively more selectively than COX-2.

• Tuberculosis and Respiratory Diseases
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• v.47 no.4
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• pp.451-459
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• 1999

Background : In sepsis, excessive generation of reactive oxygen species plays key roles in the pathogenesis of acute lung injury. The serum antioxidants such as catalase and MnSOD are elevated in sepsis and considered as predictors of acute respiratory distress syndrome(ARDS) and prognostic factors of sepsis. Peroxiredoxin(Prx) has recently been known as an unique and major intracellular antioxidant. In this study, we evaluated the expression of Prx I and Prx II in mouse monocyte-macrophage cells(RAW 267.7) after treatment of oxidative stress and endotoxin and measured the amount of Prx I, Prx II and thioredoxin(Trx) in peritoneal and bronchoalveolar lavage fluid of septic animal model. Methods : Using immunoblot analysis with specific antibodies against Prx I, Prx II and Trx, we evaluated the distribution of Prx I and Prx II in human neutrophil, alveolar macrophage and red blood cell. We evaluated the expression of Prx I and Prx II in mouse monocyte-macrophage cells after treatment of $5\;{\mu}M$ menadione and $1\;{\mu}g/ml$ lipopolysaccharide(LPS) and measured the amount of Prx I, Prx II and Trx in peritoneal lavage fluid of intraperitoneal septic animals(septic animal model induced with intraperitoneal 6 mg/Kg LPS injection) and those in bronchoalveolar lavage fluid of intraperitoneal septic animals and intravenous septic animals(septic animal model induced with intravenous 5 mg/Kg LPS injection) and compared with the severity of lung inflammation. Results : The distribution of Prx I and Prx II were so different among human neutrophil, alveolar macrophage and red blood cell. The expression of Prx I in mouse monocyte-macrophage cells was increased after treatment of $5\;{\mu}M$ menadione and $1\;{\mu}g/ml$ lipopolysaccharide but that of Prx II was not increased. The amount of Prx I, Prx II and Trx were increased in peritoneal lavage fluid of intraperitoneal septic animals but were not increased in bronchoalveolar lavage fluid of intraperitoneal and intravenous septic animals regardless of the severity of lung inflammation. Conclusion : As intracellular antioxidant, the expression of Prx I is increased in mouse monocyte-macrophage cells after treatment of oxidative stress and endotoxin. The amount of Prx I, Prx II and Trx are increased in local inflammatory site but not increased in injured lung of septic animal model.

• Journal of the Korean Dietetic Association
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• v.12 no.1
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• pp.82-88
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• 2006

This study was performed to evaluate the effect of Sorghum bicolor L. Moench(Sorghum, su-su) extracts on mouse immune cell activation. As ex vivo experiment, different concentrations(0, 50, 500mg/kg B.W.) of Sorghum bicolor L. Moench water extracts were orally administrated into mouse every other day for four weeks. The proliferation of mouse splenocytes, the number of plaque forming cells(PFC) and the cytokine IL-1β production by activated macrophage were used as indices for immunocompetence. Splenocyte proliferation was enhanced in mouse orally administrated with 50mg/kg B.W./day concentration compared to that of control group. Especially, the highest proliferation of spleoncyte was seen in the mouse orally administrated at the concentration of 50mg/kg B.W./day. The number of plaque forming cells(PFC) to SRBC were significantly enhanced when compared with control group. Also, the mouse of Sorghum bicolor L. Moench water extracts 50mg/kg B.W./day supplementation group with LPS stimulation enhanced level of IL-1$\beta$ cytokine production. This study suggest that supplementation of Sorghum bicolor L. Moench water extracts may enhance the immune function by regulating the splenocytes proliferation, increasing the number of PFC and enhancing the cytokine production by activated macrophage.

• Herbal Formula Science
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• v.5 no.1
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• pp.169-178
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• 1997

To investigate effect of water extract of Asparagi Tuber(天門冬) on human cancer cell-lines and immunocytes, this research estimated proliferation of A431 cell line, KHOS-NP cell line, mouse thymocytes and mouse splenocytes, Nitric Oxide(NO) from macrophage, apoptosis and subpopulation of the mouse thymocytes. The result were obtained as follows ; 1. Asparagi Tuber inhibited the proliferation of A431 cell line. 2. Asparagi Tuber inhibited the proliferation of KHOS-NP cell line. 3. Asparagi Tuber accelerated the proliferation of mouse thymocytes. 4. Asparagi Tuber inhibited the proliferation of mouse splenocytes. 5. Asparagi Tuber $100{\mu}g/m{\ell}$ inhibited the production of NO from macrophages in vitro, being compared NPS+IFN treated group. 6. Asparagi Tuber inhibited the production of NO from macrophages in vivo, being compared LPS+IFN treated group. 7. Asparagi Tuber accelerated the induction of apoptosis of the mouse thymocytes. 8. In subpopulation Asparagi Tuber increased $T_H$ of the mouse thymocytes, but decreased $T_C/T_S$ of the mouse thymocytes.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.5
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• pp.914-920
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• 2002

This experiment is carried out for test whether the addition temperament drugs of Danchisoyo-san have an anti-inflammatory effect and have suppression effect on immunocyte in the state of inflammation which induced by carageenan and zymosan. The freezing lyophilization powder which were extracted from Danchisoyo-san divided low dose group(200mg/kg-DSL) and high dose group(500mg/kg-DSH) and after melting in water, it was orally administered to the mouse. Compared with inflammation induced group which were induced by triggering-inflammation reagent carageenan and zymosan and normal contrast group, we measured the edema decrement effect, macrophage and spleen cell activation. 1. Danchisoyo-san has suppress inflammatory reaction induced by carrageenan. 2. Danchisoyo-san has suppress increasing activation of abdominal cavity macrophage cell in the carrageenan induced inflammation. 3. Danchisoyo-san has suppress increasing activation of spleen cell in the carrageenan induced inflammation. 4. Danchisoyo-san has suppress increasing activation of abdominal cavity macrophage in the zymosan induced inflammation. 5. Danchisoyo-san has suppress increasing activation of spleen cell in the zymosan induced inflammation. Based on the above result, Danchisoyo-san was improved its suppression effect to the inflammatory reaction through the suppression of spleen cell and macrophage activation. So we concluded that Danchisoyo-san is prospected as a anti-inflammatory agent to cure inflammation.