• Journal of Microbiology and Biotechnology
• /
• v.18 no.4
• /
• pp.686-694
• /
• 2008

The inhibitory effects of 5,6,3',5'-tetramethoxy 7,4'-hydroxyflavone (labeled as p7F) were elucidated on the productions of proinflammatory cytokines as well as inflammatory mediators in human synovial fibroblasts and macrophage cells. p7F inhibited IL-1${\beta}$ or TNF-${\alpha}$ induced expressions of inflammatory mediators (ICAM-1, COX-2, and iNOS). p7F also inhibited LPS-induced productions of nitric oxide and prostaglandin $E_2$ in RAW 264.7 cells. In order to investigate whether p7F would inhibit IL-1 signaling, p7F was added to the D10S Th2 cell line (which is responsive to only IL-1${\beta}$ and thus proliferates), revealing that p7F inhibited IL-1${\beta}$-induced proliferation of D10S Th2 cells in a dose-response manner. A flow cytometric analysis revealed that p7F reduced the intracellular level of free radical oxygen species in RAW 264.7 cells treated with hydrogen peroxide. p7F inhibited IkB degradation and NF-${\kappa}$B activation in macrophage cells treated with LPS, supporting that p7F could inhibit signaling mediated via toll-like receptor. Taken together, p7F has inhibitory effects on LPS-induced productions of inflammatory mediators on human synovial fibroblasts and macrophage cells and thus has the potential to be an anti-inflammatory agent for inhibiting inflammatory responses.

• Korean Journal of Medicinal Crop Science
• /
• v.14 no.2
• /
• pp.63-69
• /
• 2006

The methanolic (MeOH) extract of A. fruticosa bark, which showed immune-regulatory activities, was separated to purify an active compared by means of a multi-stage column chromatography. This resulted in the isolation and characterization of an isoflavone glycoside named 4', $6-Dimethoxyisoflavone-7-O-{\beta}-D-glucopyranoside$. Immuno-regulatory activities of the crude extract of Amorpha fruticosa LINNE bark were compared with that of an isoflavone glycoside (4', $6-dimethoxyisoflavone-7-O-{\beta}-D-glucopyranoside$). The crude methanolic extract of A. fruticosa and purified single compound showed 16% of relatively low cytotoxicity at a maximum concentration of 1.0 g/L in cultivated normal human lung cell line (HEL299). Cell growth of human T cells was increased up to 15%, 0.5 g/L of the crude extract added group. This was higher than a single compound added one. On the other hand, specific production rates of IL-6 and $TNF-{\alpha}$ from T cell were higher in the purified compound treat group ($0.82{\times}10^{-4}\;pg/cell$ and $1.08{\times}10^{-4}\;pg/cell$, respectively), compared to 0.5 g/L of the crude extract added group ($0.65{\times}10^{-4}\;pg/cell$ and $0.84{\times}10^{-4}\;pg/cell$, respectively). In addition, the growth of NK-92MI cells incubated with the crude extract was higher up to 56% over the cells grown with a single compound (0.5 g/L). In overall, the crude extract showed relatively higher immuno-regulatory activities compared with a single compound, probably due to the synergic effect given by other substances existed in the crude extract. Even though the siolated compound stimulated higher secretion of cytokines from human T cells.

• Natural Product Sciences
• /
• v.18 no.1
• /
• pp.60-66
• /
• 2012

Human skin is the first line of defense for the protection of the internal organs of the body from different stimuli. Ultraviolet B (UVB) irradiation induces skin damage and inflammation through the secretion of various cytokines, which are immune regulators produced by cells. To prevent the initiation of skin inflammation, keratinocytes that have been irreversibly damaged by radiation must be removed through the apoptotic mechanism. Ixeris dentata (family: Asteraceae) is a perennial medicinal herb indigenous to Korea. It has been used in Korea, China, and Japan to treat in digestion, pneumonia, diabetes, hepatitis, and tumors. To gain insight into the anti-inflammatory effects of I. dentata, we examined its influence on UVB-induced pro-inflammatory cytokine production in human keratinocytes (HaCaT cells), by observing cells that were stimulated with UVB in the presence or absence of I. dentata. In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay, reverse transcription polymerase chain reaction, and western blot analysis to measure the activation of mitogen-activated protein kinase (MAPKs). I. dentata inhibited UVBinduced production of the pro-inflammatory cytokine interleukin (IL)-6 in a dose-dependent manner. Further, I. dentata inhibited the UVB-induced expression of cyclooxygenase (COX)-2. Furthermore, I. dentata inhibited the phosphorylation of c-Jun NH2-terminal kinase and p38 MAPKs, suggesting that it inhibits the secretion of the pro-inflammatory cytokines IL-6 and IL-8, and COX-2 expression, by blocking MAPK phosphorylation. These results suggest that I. dentate can potentially protect against UVB-induced skin inflammation.

• Korean Journal of Medicinal Crop Science
• /
• v.10 no.2
• /
• pp.109-115
• /
• 2002

The immune activation activities of ethanol, ethanol : water(1 : 1,v/v) and water extracts from Dendropanax morbifera Lev. were compared. The crude extracts showed relatively low cytotoxicity as less than 26% by using human normal liver cell(WRL-68). For example, the ethanol extract inhibited. in MCF7 and Hep3B cells in adding 1.0 mg/mL ethanol extracts. The growth of human immune T and B cells was enhanced up to $1.22{\sim}1.27$ times by adding the crude extracts. These are a trend of increasing secretion of $cytokines(IL-6,\;TNF-{\alpha})$ from human B/T cell for 6 day cultivation. It was found that the immune activation activities of the crude extracts seemed to be higher than those obtained by using other solvents.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.46 no.2
• /
• pp.195-204
• /
• 2020

The purpose of this study was to investigate the effects of 14 Chung-bu medicinal materials described in the Dongui Bogam on inflammatory cytokine production in HaCaT human keratinocyte cells. In order to confirm this possibility, we screened inhibition activity of 17 cytokines using Bio-Plex Pro^TM Human Cytokine 17-plex assay in HaCaT cell lines. Of the 14 Chung-bu medicinal materials, Holotrichia (Ho) and Scorpio (Sc) exerted inhibitory effects on interleukin (IL)-5 production; Ho, Mantidis Ootheca (MO), and Hirudo (Hi) exerted inhibitory effects on IL-6 production; Ho, MO, Lumbricus (Lu), Hi, and Meretricis Concha (MC) showed significant inhibitory effects on IL-8 production; Gecko (Ge), Bombycis Faeces (BF), Cicadidae Periostracum (CP), and MC showed significant inhibitory effects on IL-13 production; and Testudinis Chinemis Plastrum et Carapax (TCPC), BF, and Lu exerted significant inhibitory effects on MIP-1β production. Results indicated that the Chung-bu medicinal materials might be a good candicate as potential anti-inflammatory agents for inhibition of skin inflammation. However, further investigations on these materials, including mechanistic studies, should be carried out to validated the effects in human skin equivalent models of dermatitis.

• Journal of Life Science
• /
• v.21 no.12
• /
• pp.1778-1783
• /
• 2011

Hematopoietic cytokines regulate production of blood cells by stimulating proliferation and differentiation of bone marrow cells. Among these hematopoietic cytokines, called hematopoitic growth factors, glranulocyte-colony stimulating Factor (G-CSF), which regulates growth of neutrophils, is one of important therapeutic factors because cancer patients suffer with neutropenia which is severe reduction of neutrophils after chemotherapy. Two groups of recombinant G-CSF have approved and used for therapeutic purposes and many researches are still on-going to produce recombinant G-CSF by different techniques. We engineered human G-CSF with Bombyx specific endoplasmic reticulum (ER) signal sequence, therefore, secretion of human G-CSF protein was improved in Bombyx mori-origined cell line, Bm5. The Bombyx ER signal sequence and human G-CSF matured protein region chimera was further remodeled at the N-terminus of matured G-CSF protein to understand roles of N-terminus on outer cellular secretion and/or production. Three different mutants were generated deleting three amino acids in non alpha-helical region in N-terminus in order to scan important amino acids for G-CSF secretion. One of 3 different N-terminal deletion mutants showed dramatically reduction of secreted amount of G-CSF indicating its important role on secretion. The data suggest that remodeling in non alpha-helical region of N-terminus is also important for recombinant G-CSF production.

• IMMUNE NETWORK
• /
• v.23 no.6
• /
• pp.48.1-48.21
• /
• 2023

Mesenchymal stromal/stem cells (MSCs) possess immunoregulatory properties and their regulatory functions represent a potential therapy for acute lung injury (ALI). However, uncertainties remain with respect to defining MSCs-derived immunomodulatory pathways. Therefore, this study aimed to investigate the mechanism underlying the enhanced effect of human recombinant bone morphogenic protein-2 (rhBMP-2) primed ES-MSCs (MSC^BMP2) in promoting Tregs in ALI mice. MSC were preconditioned with 100 ng/ml rhBMP-2 for 24 h, and then administrated to mice by intravenous injection after intratracheal injection of 1 mg/kg LPS. Treating MSCs with rhBMP-2 significantly increased cellular proliferation and migration, and cytokines array reveled that cytokines release by MSC^BMP2 were associated with migration and growth. MSC^BMP2 ameliorated LPS induced lung injury and reduced myeloperoxidase activity and permeability in mice exposed to LPS. Levels of inducible nitric oxide synthase were decreased while levels of total glutathione and superoxide dismutase activity were further increased via inhibition of phosphorylated STAT1 in ALI mice treated with MSC^BMP2. MSC^BMP2 treatment increased the protein level of IDO1, indicating an increase in Treg cells, and Foxp3⁺CD25⁺ Treg of CD4⁺ cells were further increased in ALI mice treated with MSC^BMP2. In co-culture assays with MSCs and RAW264.7 cells, the protein level of IDO1 was further induced in MSC^BMP2. Additionally, cytokine release of IL-10 was enhanced while both IL-6 and TNF-α were further inhibited. In conclusion, these findings suggest that MSC^BMP2 has therapeutic potential to reduce massive inflammation of respiratory diseases by promoting Treg cells.

• Korean Journal of Medicinal Crop Science
• /
• v.13 no.1
• /
• pp.41-47
• /
• 2005

This study was performed to experiment about useful biological activities of the parts of the extracts from Amorpha fruticosa LINNE. Experimental studies were progressed through the anticancer activities and immune activities such as cell cytotoxicity, inhibition activities of cell growth, cell growth of human B and T cell, productivity of cytokines and natural killer cell activities. The cell cytotoxicity using human Lung normal cell (HEL299) was showed cytotoxicity of below 22% by extracts of Amorpha fruticosa L. in 1.0 g/l concentration. The anticancer activities were increased in over 70% by roots extracts of Amorpha fruticosa L. in A549 and AGS cells. The immune cell growth using human immune B and T cells was increased against control by barks extracts of Amorpha fruticosa L. The secretion of cytokines $(IL-6,\;TNF-{\alpha})$ from human immune B and T cells was showed secretion of the amount of cytokines by roots extracts of Amorpha fruticosa L. Also NK cell growth was increased against control all of the extracts of Amorpha fruticosa L. From the results, the roots and barks extracts of Amorpha fruticosa L. were showed useful biological activities.

• BMB Reports
• /
• v.43 no.8
• /
• pp.541-546
• /
• 2010

We utilized a mammalian expression system to purify and characterize autotaxin (ATX)/lysophospholipase D, an enzyme present in the blood responsible for biosynthesis of lysophosphatidic acid. The human ATX cDNA encoding amino acids 29-915 was cloned downstream of a secretion signal of CD5. At the carboxyl terminus was a thrombin cleavage site followed by the constant domain (Fc) of IgG to facilitate protein purification. The ATX-Fc fusion protein was expressed in HEK293 cells and isolated from conditioned medium of a stable clone by affinity chromatography with Protein A sepharose followed by cleavage with thrombin. The untagged ATX protein was further purified to essential homogeneity by gel filtration chromatography with a yield of approximately 5 mg/liter medium. The purified ATX protein was enzymatically active and biologically functional, offering a useful tool for further biological and structural studies of this important enzyme.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.6
• /
• pp.2807-2812
• /
• 2012

The purpose of this study was to examine the effect of a Toll-like receptor 5 (TLR5) agonist, CBLB502, on the growth and radiosensitivity of A549 lung cancer cells in vivo. Expression of myeloid differentiation factor 88 (MyD88) or TLR5 was stably knocked down in human lung cancer cells (A549) using lentivirus expressing short hairpin RNA targeting human MyD88 or TLR5. Lack of MyD88 or TLR5 expression enhanced tumor growth in mouse xenografts of A549 lung cancer cells. CBLB502 inhibited the growth of A549 lung cancer cells, not A549-MyD88-KD cells in vivo in the murine xenograft model. Our results showed that the inhibition of A549 by CBLB502 in vivo was realized through regulating the expression of neutrophil recruiting cytokines and neutrophil infiltration. Finally, we found that activation of TLR5 signaling did not affect the radiosensitivity of tumors in vivo.