• The Korea Journal of Herbology
• /
• v.32 no.6
• /
• pp.9-15
• /
• 2017

Objectives : Traditional medicines isolated from natural products often have positive effects in the prevention and healing of various immune disorders, such as allergy and atopic inflammation. Scrophulariae Radix (SR) been used in oriental medicine used for treatment of acute and chronic inflammatory diseases. Mast cells are known to play important roles in the initiation of allergic reactions. In this study, we investigated the effects of SR ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : Rat basophilic leukemia RBL-2H3 cells were purchased from Korean Cell Line Bank (KCLB No. 22256). Cell viability was measured by MTT assay. Assays for ${\beta}-Hexosaminidase$ Secretion : RBL-2H3 cells were sensitized with dinitrophenyl-ImmunoglobulinE (DNP IgE). The next antigen DNP-BSA ($25ng/m{\ell}$) was added for 10 minutes and the reaction was terminated after 5 minutes in the ice bath. To determine ${\beta}-Hexosaminidase$ release, supernatants were aliquoted into 96-well plates. Samples were mixed with substrate solution and incubated for 1 h at $37^{\circ}C$. Absorbance was measured with a spectrophotometer at 405 nm. IL-4 and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) concentrations in cell culture supernatants were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results : The cytotoxicity of SRE in RBL-2H3 cells was less than 5%. SRE inhibited DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. Also significantly decreased the levels of inflammatory cytokine, IL-4 and TNF-alpha. In this study, the SRE showed potential anti-allergic and antiinflammatory. Conclusions : These results indicate that SRE could be inhibit the allergic response through suppressing the mast cell activation.

• The Korea Journal of Herbology
• /
• v.32 no.2
• /
• pp.107-114
• /
• 2017

Objectives : Tribulus terrestris $Linn{\acute{e}}$ (Tribuli Fructus; TF) has been used to treat hypochondrium, agalactia, nebula, itching and vitiligo in traditional Korean medicine. In this study, we investigated the effects of TF 30% ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : TF extract was prepared by 30% ethanol. RBL-2H3 cells, a rat mast cell line, were treated with TF extract at different concentrations for 1 hr and then stimulated with DNP-IgE/HSA for indicated times. Cell viability was measured by WST-1 assay. The expression of inflammatory cytokines (IL-4, IL-13 and $IFN-{\gamma}$) mRNA was determined by reverse transcriptase-PCR, and the phosphorylation of ERK1/2, p38 and JNK MAP kinases (MAPKs) was determined by Western blot. The nuclear expression of $NF-{\kappa}B$ p65 in the cells was detected by Western blot and immunocytochemistry, respectively. Results : The treatment of TF extract at 0.1 and $0.2mg/m{\ell}$ significantly decreased the expression of IL-4 and IL-13 mRNA in IgE-stimulated RBL-2H3 mast cells, while significantly increased the expression of $IFN-{\gamma}$ mRNA. TF extract treatment was also inhibited the phosphorylation of ERK1/2, p38 and JNK MAPKs in IgE-stimulated RBL-2H3 mast cells in a dose-dependent manner. In addition, TF extract significantly blocked the translocation of $NF-{\kappa}B$ p65 into the nuclear of cells after IgE stimulation. Conclusions : These results indicate that TF extract inhibits inflammatory response in IgE-stimulated mast cells through blocking MAPKs/$NF-{\kappa}B$ pathway. This suggests that TF extract has an anti-inflammatory activity in mast cell activation.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.19 no.6
• /
• pp.1647-1655
• /
• 2005

Gamimahaenggamsuk-tang (GMHGST) is used for treatment of inflammatory diseases including rheumatoid arthritis (RA). Here, regulatory activity of GMHGST on RA-mediated inflammatory responses was investigated in cultured human fiDroblast-like synoviocytes (FLS), Levels of mRNAs encoding for inflammatory cytokines such as $IL-1{\beta}$, IL-6 and IL-8 and NOS-II enzyme, which had been induced by $TNF-{\alpha}$ and $IL-1{\beta}$ cotreatment, were decreased to the similar levels as those in cells treated with anti-inflammatory agent MTX. mRNA expressions of matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinases (TIMPs) as well as intercellular adhesion molecule (ICAM) were also downregulated by increasing doses of GMHGST in activated FLS. Moreover, GMHGST appeared to protect cells by decreasing NO levels, and inhibited cell proliferation which had been induced by inflammatory stimulation by $TNF-{\alpha}$ and IL-1. These results suggest that GMHGST is effective as an inhibitory agent for regulating inflammatory responses in activated FLS.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.25 no.6
• /
• pp.982-988
• /
• 2011

This research seeks a basis for developing new anti-inflammatory medicine by investigating Cornus Walteri extract for its anti-inflammatory effects. After the injection of LPS in to rats with Cornus Walteri extract, its anti-inflammatory effects were compared among the treatment groups. The plasma concentration of IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ peaked at 5h after LPS injection, and the values of the Cornus Walteri extract groups were lower than those of the control group. In the increment of concentration of these cytokines at 2h and 5h after LPS injection, the Cornus Walteri groups were lower than that of control group. The plasma concentration of IL-10 peaked at 5h after LPS injection, and the values of the Cornus Walteri extract groups were higher than those of the control group. In the increment of cytokines concentration at 2h and 5h after LPS injection, the Cornus Walteri groups were higher than that of control group. Liver cytokines measurement was done at 5h after LPS injection. The concentration of liver IL-$1{\beta}$ and IL-6 in the Cornus Walteri groups was lower than that of the control group. The concentrations of liver TNF-${\alpha}$, and IL-10 showed no significant differences among all the treatment groups. In the studies of lipopolysaccharide-exposed Raw 264.7 cells, the concentration of IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ in the lipopolysaccharide-exposed cells groups was higher than that of control group (normal group). However, in lipopolysaccharide-exposed cells groups, they showed lower values than those of control group and these values showed a tendency to decrease in the Cornus Walteri groups. The concentration of IL-10 in the lipopolysaccharide-exposed cells groups was higher than that of control group (normal group), and among the lipopolysaccharide-exposed cells groups, all Cornus Walteri extract groups showed higher values than single lipopolysaccharide-exposed cells groups. This studies have shown that in vitro and in vivo Cornus Walteri extracts are significantly more sensitive to inflammatory cytokines and LPS induced lethality. We conclude that the Cornus Walteri extracts have an functional material for inflammatory activities.

• Korean Journal of Acupuncture
• /
• v.39 no.2
• /
• pp.43-53
• /
• 2022

Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.

• Journal of Korean Medicine Rehabilitation
• /
• v.32 no.4
• /
• pp.9-18
• /
• 2022

Objectives This study was conducted to confirm the anti-inflammatory effect of Naetakbaekryeom-san (NTB), and whether it could be another treatment for inflammatory diseases. Methods The NTB water extract was extracted with hot water at 100℃ for 2 hours, concentrated at 80℃ under reduced pressure, and used. After 2 hours of pretreatment with NTB and positive control Bay11-7082, nitric oxide (NO), inducible NO synthase (iNOS), interleukin (IL)-6, IL-1𝛽, tumor necrosis factor alpha (TNF-𝛼) were measured in RAW264.7 cells activated with lipopolysaccharides (LPS) 500 ng/mL. After 2 hours of pretreatment with NTB, the anti-inflammatory effect of NTB was evaluated by measuring nuclear factor kappa-light-chain-enhancer of activated B cells (NF-𝜅B) in RAW264.7 cells and 293T cells activated with phorbol 12-myristate 13-acetic acid (PMA) 30 ng/mL. Results In RAW264.7 cells activated with LPS, NTB at concentrations of 0.1, 0.3, and 1.0 mg/mL showed no cytotoxicity, significantly inhibited NO production and inhibition of iNOS expression. TNF-𝛼 cytokine levels was not regulated, but NTB at each concentration inhibited the production of IL-1𝛽 and IL-6, and the effect was higher than that of the positive control Bay11-7082 (20 𝜇M). In PMA-activated RAW264.7 cells and 293T cells, each concentration of NBT decreased the NF-𝜅B transcriptional activity, with the greatest decrease at 1 mg/mL. Conclusions These results demonstrated the anti-inflammatory effect of NTB water extracts, but further studies such as comparison of anti-inflammatory effects and antioxidant effects by NTB component, comparison of effects according to extraction solvents, and clinical studies are needed.

• IMMUNE NETWORK
• /
• v.14 no.1
• /
• pp.30-37
• /
• 2014

Collaboration of TLR and non-TLR pathways in innate immune cells, which acts in concert for the induction of inflammatory cytokines, can mount a specific adaptive immune response tailored to a pathogen. Here, we show that murine DC produced increased IL-23 and IL-6 when they were treated with LPS together with curdlan that activates TLR4 and dectin-1, respectively. We also found that the induction of the inflammatory cytokine production by LPS and curdlan requires activation of IKK. However, the same treatment did not induce DC to produce a sufficient amount of TGF-${\beta}$. As a result, the conditioned media from DC treated with LPS and curdlan was not able to direct $CD4^+$ T cells to Th17 cells. Addition of TGF-${\beta}$ but not IL-6 or IL-$1{\beta}$ was able to promote IL-17 production from $CD4^+$ T cells. Our results showed that although signaling mediated by LPS together with curdlan is a potent stimulator of DC to secrete many pro-inflammatory cytokines, TGF-${\beta}$ production is a limiting factor for promoting Th17 immunity.

• The Journal of Internal Korean Medicine
• /
• v.25 no.1
• /
• pp.46-58
• /
• 2004

Objectives : This study was designed to investigate the effects of Injinchunggan-tang(Yinchenqinggan-tang) on the expression of inflammatory cytokine genes and proteins in kupffer cells. Materials and Methods : The mRNA expression level and protein secretion level were measured using quantitative RT-PCR and ELISA assay respectively in Injinchunggan-tang-treated and untreated kupffer cells after exposed to ethanol, acetaldehyde and lipopolysaccharide. Results : Injinchunggan-tang(Yinchenqinggan-tang) reduced mRNA expression level and protein secretion level of $TNF-{\alpha},\;TGF-{\beta}1,\;IL-1{\beta},\;IL-6,\;IL-8$ that are induced by ethanol, acetaldehyde and lipopolysaccharide in kupffer cells and that mediate inflammation and fibrosis of liver. Conclusion : The result indicates that Injinchunggan-tang (Yinchenqinggan-tang) blocks alcohol-induced liver injury and protects liver by reducing production of inflammatory cytokines.

• The Journal of Pediatrics of Korean Medicine
• /
• v.21 no.2
• /
• pp.97-107
• /
• 2007

Objectives This experiment is about Jojoongikgi-tang(JIT) exerts anti-inflammatory effects in BV2 microglial cells, and the effect of JIT on Nitric oxide(NO) production in lipopolysaccharide(LPS)-stimulated BV2 microglial cells were also demonstrated. Methods To investigate the anti-inflammatory effects of JIT, NO production, expression level of iNOS mRNA, PGE2 synthesis, expression COX-2 mRNA, cell viability, $TNF-{\alpha}$ mRNA expression were examined. Results The expression level of inducible nitric oxide synthase(iNOS) was decreased by JIT, and the production of Prostaglandin E2(PGE2) and the expression of Cox-2 mRNA also were inhibited by JIT. Proinflammatory mediators, such as $TNF-{\alpha}$, $IL-1{\beta}$, IL-12, were inhibited by JIT in a dose-dependent manner. Conclusions JIT have anti-inflammatory effects in BV2 microglial cells and could be used in inflammatory disease.

• Applied Biological Chemistry
• /
• v.49 no.1
• /
• pp.65-69
• /
• 2006

With extracted Aloe vera and Aloe arborescens, anti-inflammatory effect was examined in phosphatidic acid-stimulated Raw 264.7 cells. Phosphatidic acid $(50\;{\mu}g/ml)$ treatment increased inflammation related 264.7 cells. The extracts of $80\;{\mu}g/ml$ concentration from Aloe arborescens reduced the expression of iNOS. Aloe vera and Aloe arborescens also reduced the expression of COX-2, it seems that anti-inflammatory effects of Aloe extracts is partly due to the inhibition of COX-2 expression by inhibiting in Raw 264.7 cells.