• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2020년도 춘계학술대회
• /
• pp.87-87
• /
• 2020

In this study, we investigated the immune-enhancing activity of water extracts from Hydrangea macrophylla subsp. serrata (WE-HML). WE-HML increased cell viability and production of immunomodulators, which contributed to activating phagocytic activity in RAW264.7 cells. Inhibition of JNK and NF-κB reduced the production of immunomodulators by WE-HML. ROS inhibition suppressed the production of immunomodulators, and the activation of JNK and NF-κB signaling by WE-HML. TLR4 inhibition attenuated the production of immunomodulators, and activation of JNK and NF-κB signaling by WE-HML. In the immunosuppressed mouse model, WE-HML increased the spleen index, the levels of the cytokines, the numbers of white blood cells, lymphocytes, and neutrophils. However, WE-HML inhibited LPS-mediated overproduction of pro-inflammatory mediators in RAW264.7 cells, which indicated that WE-HML may have anti-inflammatory activity under excessive inflammatory conditions. Taken together, WE-HML may be considered to have immune-enhancing activity and expected to be used as a potential immune-enhancing agent.

• Journal of Ginseng Research
• /
• 제39권4호
• /
• pp.365-370
• /
• 2015

Background: The beneficial effects of ginsenoside species have been well demonstrated in a number of studies. However, the function of ginsenoside Ro (GRo), an oleanane-type saponin, has not been sufficiently investigated. Thus, the aim of the present study was to investigate the anti-inflammatory effects of GRo in vitro using the Raw 264.7 mouse macrophage cell line treated with lipopolysaccharide (LPS), and to clarify the possible mechanism of GRo involving heme oxygenase-1 (HO-1), which itself plays a critical role in self-defense in the presence of inflammatory stress. Methods: Raw 264.7 cells were pretreated with GRo (up to $200{\mu}M$) for 1 h before treatment with 1 mg/mL LPS, and both cell viability and inflammatory markers involving HO-1 were evaluated. Results: GRo significantly increased cell viability in a dose dependent manner following treatment with LPS, and decreased levels of reactive oxygen species and nitric oxide. GRo decreased inflammatory cytokines such as nitric oxide synthase and cyclooxygenase-2 induced by LPS. Moreover, GRo increased the expression of HO-1 in a dose dependent manner. Cotreatment of GRo with tin protoporphyrin IX, a selective inhibitor of HO-1, not only inhibited upregulation of HO-1 induced by GRo, but also reversed the anti-inflammatory effect of GRo in LPS treated Raw 264.7 cells. Conclusion: GRo induces anti-inflammatory effects following treatment with LPS via upregulation of HO-1.

• 한국자원식물학회지
• /
• 제34권3호
• /
• pp.203-215
• /
• 2021

Bok choy is one of Brassica vegetables widely consumed worldwide. Brassica vegetables have been reported to exert various pharmacological activities such as antioxidant, anti-cancer and cardioprotective activity. However, studies on immunostimulatory activity of bok choy sprout have not been conducted properly. Thus, in this study, we investigated in vitro immunostimulatory activity of bok choy sprout extract (BCS) using mouse macrophage RAW264.7 cells. Our results showed that BCS increased the production of immunomodulators such as NO, iNOS, IL-1β, IL-6, IL-12, TNF-α and MCP-1, and phagocytic activity in RAW264.7 cells. BCS activated MAPK, NF-κB and PI3K/AKT signaling pathways. However, BCS-mediated production of immunomodulators was dependent on JNK, NF-κB and PI3K/AKT signaling pathways. the mRNA expression of TLR2 were significantly increased by BCS, TLR2 inhibition by anti-TLR2 dramatically suppressed the production of immunomodulators by BCS. In addition, TLR2 inhibition by anti-TLR2 significantly reduced BCS-mediated phosphorylation level of AKT, JNK and NF-κB. From these results, BCS may have immunostimulatory activity via TLR2-MAPK, NF-κB and PI3K/AKT signaling pathways. Therefore, BCS expected to be used as a potential immune-enhancing agent.

• 대한본초학회지
• /
• 제27권4호
• /
• pp.99-107
• /
• 2012

Objectives : The aim of this study was to authenticate whether fractionated extract of Rumex japonicus HOUTT. (RJ) has anti-inflammatory effects in mouse macrophage, RAW264.7 cells. Methods : Roots of RJ were extracted by methanol for 48hours. The methanol that gained was filtered and freeze dried. The methanol extract was dissolved in water and dichloromethane (DCM). After that, two layers were separated. Ethyl acetate (EA) added to the water layer and separated again. All the layers were filtered and freeze dried and the extracts were tested. Cytotoxic activity of extracts on RAW 264.7 cells was measured using MTS assay. The nitric oxide (NO) production was measured and proinflammatory cytokines and $PGE_2$ were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), I ${\kappa}$-B-${\alpha}$ and nuclear NF-${\kappa}B$ p65 expression were detected by western blot. Results : Our results indicated that DCM and EA extracts of RJ inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 production in RAW 264.7 cells most effectively. DCM and EA extracts also had suppression effects of LPS-induced NF-${\kappa}B$ and MAPKs activation. Conclusions : This results demonstrate that fractionated extract of RJ has anti-inflammatory effects and among the fractioned extract, dichloromethane and ethyl acetate extract have best anti-inflammatory effects.

• The Korean Journal of Physiology and Pharmacology
• /
• 제27권3호
• /
• pp.257-265
• /
• 2023

Kidney ischemia/reperfusion (I/R) injury, a common cause of acute kidney injury (AKI), is associated with the migration of inflammatory cells into the kidney. Ras-related C3 botulinum toxin substrate 1 (Rac1), a member of the Rho family of small GTPase, plays an important role in inflammatory cell migration by cytoskeleton rearrangement. Here, we investigated the role of Rac1 on kidney I/R injury and macrophage migration. Male mice were subjected to either 25 min of bilateral ischemia followed by reperfusion (I/R) or a sham operation. Some mice were administrated with either NSC23766, an inhibitor of Rac1, or 0.9% NaCl (vehicle). Kidney damage and Rac1 activity and expression were measured. The migration and lamellipodia formation of RAW264.7 cells, mouse monocyte/macrophage, induced by monocyte chemoattractant protein-1 (MCP-1, a chemokine) were determined using transwell migration assay and phalloidin staining, respectively. In sham-operated kidneys, Rac1 was expressed in tubular cells and interstitial cells. In I/R-injured kidneys, Rac1 expression was decreased in tubule cells in correlation with the damage of tubular cells, whereas Rac1 expression increased in the interstitium in correlation with an increased population of F4/80 cells, monocytes/macrophages. I/R increased Rac1 activity without changing total Rac1 expression in the whole kidney lysates. NSC23766 administration blocked Rac1 activation and protected the kidney against I/R-induced kidney damage and interstitial F4/80 cell increase. NSC23766 suppressed monocyte MCP-1-induced lamellipodia and filopodia formation and migration of RAW 264.7 cells. These results indicate Rac1 inhibition protects the kidney against I/R via inhibition of monocytes/macrophages migration into the kidney.

• 한국자원식물학회지
• /
• 제27권3호
• /
• pp.209-214
• /
• 2014

In this study, we investigated whether A. distichum decreases the production of inflammatory mediators through downregulation of the NF-${\kappa}B$ and ERK pathway. Our data indicated that A. distichum leaf inhibits the overexpression of iNOS in protein and mRNA levels, and subsequently blocked LPS-mediated NO overproduction in RAW264.7 cells. A. distichum leaf inhibited $I{\kappa}B-{\alpha}$ degradation and p65 nuclear translocation, and subsequently suppressed transcriptional activity of NF-${\kappa}B$ in LPS-stimulated RAW264.7 cells. In addition, A. distichum leaf suppressed LPS-induced ERK1/2 activation by decreasing phosphorylation of ERK1/2. These findings suggest that A. distichum leaf shows anti-inflammatory activities through suppressing ERK-mediated NF-${\kappa}B$ activation in mouse macrophage.

• 대한한의학방제학회지
• /
• 제20권2호
• /
• pp.65-82
• /
• 2012

Objectives : The aim of this study was verification of the anti-oxidative and anti-inflammatory effects of Odukhwan(ODH) and Sasinhwan(SSH) in mouse macrophage, RAW264.7 cells. Methods : To investigate the anti-oxidative effect and scavenging activities of DPPH radical, superoxide anions, nitric oxide and peroxynitrite were measured. Cytotoxic activity of extract of ODH and SSH on RAW264.7 cells was measured using MTS assay. To proof the reductive activity of intracellular oxidation, DCFH-DA assay was performed. The nitric oxide(NO) production was measured and pro-inflammatory cytokines and $PGE_2$ were measured by ELISA kit. The levels of inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2) and nuclear NF-${\kappa}B$ p65 expression were detected by western blot. Results : After those analyses, we bring to a conclusion as follows. Both herbal formulations scavenged DPPH radical and nitric oxide. But ODH had no scavenging activity of superoxide anions and SSH had low scavenging activity in peroxynitrite. And the results indicated that ODH and SSH inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of IL-$1{\beta}$ and IL-6 production in RAW264.7 cells. They also have suppression effects of LPS-induced NF-${\kappa}B$ activation. Conclusions : ODH and SSH have anti-oxidative and anti-inflammatory effects and they may be a part of database for development of new anti-oxidative and anti-inflammatory drugs.

• Current Optics and Photonics
• /
• 제8권5호
• /
• pp.441-455
• /
• 2024

This study objective was to evaluate the effects and mechanisms of low-level laser therapy in H₂O₂-induced cell death in mouse macrophage RAW264.7 cell. After irradiation with 630 and 850 nm wavelength diode lasers with an intensity of 10 mW/cm² in RAW264.7 cells treated with 0.7 Mm H₂O₂, the effects and mechanisms of the two wavelengths on cell death inhibition were evaluated using MTT assay, ROS staining, TUNEL assay, flow cytometry analysis, and Western blot analysis. As a result, 630 or 850 nm light-emitting diodes (LED) were irradiated for 10 or 40 minutes to increase cell viability with H₂O₂ by about 1.7- or 1.6-fold, respectively. In addition, irradiation with two LEDs showed significant ROS scavenging effects, and TUNEL-positive cells were significantly reduced by 45.7% (630 nm) and 37.8% (850 nm) compared to cells treated with H₂O₂ alone. The Bax/Bcl-2 ratio of cells irradiated with both LEDs was significantly lower than that of cells treated with H₂O₂ only, and the expression of procaspase-3 and cleaved PARP was also significantly expressed in the direction of suppressing cell death. In conclusion, ROS scavenging activity by both LEDs irradiation leads to the expression of cell death pathway proteins in the direction of inhibiting cell death.

• IMMUNE NETWORK
• /
• 제10권6호
• /
• pp.230-238
• /
• 2010

Background: The MK1 strain, a novel bacterial isolate from soft-rotten tissue of the Neungee mushroom, produces copious amounts of exopolysaccharide (EPS) in a dextrose minimal medium. This study examined the molecular characteristics and immunomodulatory activity of MK1 EPS. Methods: The EPS in the culture supernatant was purified by cold ethanol precipitation, and characterized by SDS- PAGE/silver staining and Bio-HPLC. The immunomodulatory activities of the EPS were examined using the mouse monocytic cell line, RAW 264.7 cells. Results: The molecular weights of the purified EPS were rather heterogeneous, ranging from 10.6 to 55 kDa. The EPS was composed of glucose, rhamnose, mannose, galactose, and glucosamine at an approximate molar ratio of 1.00 : 0.8 : 0.71 : 0.29 : 0.21. EPS activated the RAW cells to produce cytokines, such as TNF-${\alpha}$ and IL-$1{\beta}$, and nitric oxide (NO). EPS also induced the expression of co-stimulatory molecules, such as B7-1, B7-2 and ICAM-1, and increased the phagocytic activity. The macrophage-activating activity of EPS was not due to endotoxin contamination because the treatment of EPS with polymyin B did not reduce the macrophage-activating activity. Conclusion: The EPS produced from the MK1 strain exerts macrophage-activating activity.

• 동의생리병리학회지
• /
• 제21권3호
• /
• pp.765-770
• /
• 2007

Forsythiae fructus has traditionally been used for the treatment of erysipelas, skin rash and acute or chronic inflammatory disorders. The effect of Forsythiae fructus against lipopolysaccharide-induced inflammation was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcription-polymerase chain reaction (RT-PCR), on mouse RAW 264.7 macrophages. Forsythiae fructus extract suppressed the expression of iNOS, COX-2 and NF-$_K$B mRNAs on the lipopolysaccharide-stimulated enhancement in RAW 264.7 macrophages. We examined the expression of iNOS and COX-2 in both mRNA and protein levels to investigate the mechanism by which Forsythiae fructus extract inhibits NO production. Forsythiae fructus extract significantly reduced iNOS, NF-$_K$B and PGE$_2$, but didn't inhibit COX-2 expression which was induced by LPS treatment in Raw 264.7 cells. These results suggest that Forsythiae fructus exerts anti-inflammatory effects probably by suppression of the iNOS and NF-$_K$B expressions.