Transcription factors Nrf2 and NF-${\kappa}B$ are important regulators of the innate immune response, and their cross-talks in inflammation have been reported. Previously, we demonstrated that gold(I)-compound auranofin, an inhibitor of NF-${\kappa}B$ signal, induced Nrf2 activation in human synovial cells and monocytic cells. To investigate whether the Nrf2 activation is involved in the mechanism of the auranofin-attenuated NF-${\kappa}B$ signaling, we examined the effects of Nrf2 knockdown on NF-${\kappa}B$ activation using rheumatic synovial cells. When the cells were transfected with a specific siRNA for Nrf2, the gene expression was perfectly blocked. However, the Nrf2 knockdown did not cancel the suppressive effect of auranofin on TNF-$\alpha$-induced $I{\kappa}B-{\alpha}$ degradation. Treatment with a specific siRNA for HO-1, which is a target of Nrf2 and plays a role in anti-inflammation, also did not affect the blocking activity of auranofin on $I{\kappa}B-{\alpha}$ degradation. In addition, auranofin-inhibited ICAM-1 expression was not restored by Nrf2 knockdown. These findings indicate that the activated Nrf2 and HO-1 are not associated with the suppressive action of auranofin on the pro-inflammatory cytokines-stimulated NF-${\kappa}B$ activation. This suggests that Nrf2/HO-1 and NF-${\kappa}B$ signals, which are regulated by auranofin, participate in the anti-inflammatory action of auranofin via independent pathways in rheumatic synovial cells.
Allergic contact dermatitis (ACD) is an inflammatory skin disease and regarded as a prototype of T-cell mediated delayed-type hypersensitivity reactions. Poly-${\gamma}$-glutamic acid (PGA) is a biodegradable polymer that is produced by Bacillus subtilis. This study was performed to assess the effects of PGA in a canine model of ACD. ACD was induced on the back of dogs induced by sensitization and repeated application by 2,4-dinitro-1-chlorobenzene (DNCB). Topical treatment of PGA was applied once a day for 12 days and skin biophysical parameters including transepidermal water loss (TEWL), skin hydration, skin pH, skin thickness and erythema index, were measured every two days during experimental periods. Histopathology and immunohistochemistry were performed to evaluate the antiinflammatory effect. In skin biophysical parameters, TEWL, skin hydration, skin thickness and erythema index were significantly increased, with a maximum increase appeared on day 2 (p < 0.05). On the other hand, skin pH was significantly decreased, with a maximum decrease appeared on day 2 (p < 0.01). After the completion of PGA treatment, skin biophysical parameters were significantly reached those of baseline in a time-dependent manner (p < 0.05). In histopathology, marked increases of epidermal thicknesses were induced after DNCB challenge with numerous inflammatory cell infiltrations and edematous changes, decreases of connective tissue occupied regions in dermis. In addition, marked increases of cytokine - tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$)-immunoreactivities in the dermis and of apoptotic markers - caspase-3 and PARP-immunoreactivities in the epidermis were observed in DNCB-PBS control as compared with intact control, respectively (p < 0.01). It means, the ACD and related apoptotic changes were induced by DNCB in the present study. However, these ACD induced by DNCB and related apoptosis in epidermis were significantly inhibited by treatment of PGA treated skin, the decreases of infiltrated inflammatory cells and related decreases of pro-inflammatory cytokine immunoreactivities were also observed (p < 0.01). Based on these findings, PGA may have anti-inflammatory and alleviatory effects in the allergic contact dermatitis.
Ha, Eun-Suk;Hwang, Soo-Hyun;Shin, Kwang-Soon;Yu, Kwang-Won;Lee, Keyung-Ho;Choi, Joo-Sun;Park, Woo-Mun;Yoon, Taek-Joon
Korean Journal of Food Science and Technology
/
v.35
no.6
/
pp.1209-1215
/
2003
This study was carried out to establish a quantitative analysis method of separating immuno-activating substance (EN-SP) from Acanthopanax senticosus (A. senticosus) by competitive direct ELISA. Mouse antiserum (anti-EN-SP) against EN-SP was generated by immunization (s.c.) of EN-SP purified from A. senticosus as an immunogen. The titer of anti-EN-SP was about 1 : 400, and the optimal dilution of EN-SP-HRP conjugate was 1 : 1,000. When the standard curve was constructed by ELISA, its sensitivity was about $0.2{\mu}g/mL$. The coefficient variation of intra- and inter-assay were $6.13{\sim}8.81%$ and $6.73{\sim}8.60%$, respectively. According to the standard curve, the concentration of EN-SP in various senticosus extracts was found to be only $59.85\;{\mu}g$ in 10mg of extract from the bark of A. senticosus. Similarly, the immunostimulating activity to produce $TNF-{\alpha}$ or IL-12 among the various extracts of Acanthopanax was shown to be correlated with the content of EN-SP. These results demonstrated that competitive ELISA was a convenient, fast, reproducible, and accurate method for the determination of EN-SP as an immunologically active standard substance in extract of A. senticosus.
The suppression of neuroinflammatory responses in microglial cells, well known as the main immune cells in the central nervous system (CNS), are considered a key target for improving the progression of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease. Teleogryllus emma is widely consumed around the world for its broad-spectrum therapeutic effect. In a previous work, we performed transcriptome analysis on T. emma in order to obtain the diversity and activity of its antimicrobial peptides (AMPs). AMPs are found in a variety of species, from microorganisms to mammals. They have received much attention as candidates oftherapeutic drugs for the treatment of inflammation-associated diseases. In this study, we investigated the anti-neuroinflammatory effect of Teleogryllusine (VKWKRLNNNKVLQKIYFVKI-NH2) derived from T. emma on lipopolysaccharide (LPS) induced BV-2 microglia cells. Teleogryllusine significantly inhibited nitric oxide (NO) production without cytotoxicity, and reducing pro-inflammatory enzymes expression such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). In addition, Telegryllusine also inhibited the expression of pro-inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) through down-regulation of the mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling pathway. These results suggest that T. emma-derived Teleogryllusine could be a good source of functional substances that prevent neuroinflammation and neurodegenerative diseases.
Journal of the Korea Academia-Industrial cooperation Society
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v.13
no.8
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pp.3561-3569
/
2012
This study investigated the effect of mulberry fruit extract(MFE) on antioxidant and anti-inflammatory activities of middle aged women with rheumatoid factor (RF). Thirty two middle-aged subjects were divided into two groups which were normal middle-aged group (NMG) and abnormal middle-aged group whose serum RF level were > 10 u/mL (AMG). All groups had consumed MFE (100 mL/day) for 4 weeks. Anthropometric measurements, serum inflammatory factors, serum oxidative stress markers analyses were performed at baseline and then at 4 weeks following the study. There were no significant differences in anthropometric measurements, including BMI, WHR and body fat composition between two groups. But after 4 weeks MFE consumption, serum levels of C-reactive protein (CRP), ferric-reducing ability of plasma (FRAP), serum TNF-${\alpha}$, IL-2, IL-4 had significantly decreased (p<0.05) in AMG. These findings suggested that the MFE consumption as food may be protective against oxidation and inflammation like RA.
Journal of the Society of Cosmetic Scientists of Korea
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v.49
no.4
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pp.313-321
/
2023
Platycodon grandiflorus (P. grandiflorus) flower is a perennial plant belonging to the family Campanulaceae and has many excellent pharmacological effects, so it has been used as a medicinal ingredient since ancient times. In addition, anthocyanin is a purple or blue natural pigment contained in plant flowers and fruits, and is known as a powerful antioxidant. The purpose of this study was to confirm the dermatological functionality of P. grandiflorus flower extract and the value of the bluish anthocyanin contained in flowers as a cosmetic material as a natural pigment. Firstly, 50% ethanol and 80% ethanol were added to the P. grandiflorus flower and extracted under reflux for 4 h at 25, 60, and 80 ℃, and the pH of each treatment group was similar. Based on the anthocyanin content and chromaticity (E*ab), 50% ethanol 60 ℃ extraction conditions showing the color development most similar to the natural color of the P. grandifloras flower were selected, and a sample was prepared by concentrating and lyophilizing. The analysis results showed that the total phenol, total flavonoid, and total anthocyanin contents were in the ranges of 23 ㎍/mL, 16 ㎍/mL, and 0.17 ㎍/mL, respectively. The P. grandiflorus flower extract suppressed the production of nitric oxide (NO) and interleukin-6 (IL-6) in lipopolysaccharide (LPS) induced RAW264.7 cells. Furthermore, the P. grandiflorus flower extract showed wound healing effects through the promotion of skin cell migration in TNF-α stimulated human keratinocytes. The stability of anthocyanin and extract color was studied during a storage period of 50 days at various temperatures (4 ℃, 25 ℃, and 45 ℃). Color values (L, a, and b) of the P. grandiflorus flower extract changed over 50 days, whereas the bluish-purple color of the extract was stabilized using 5% maltodextrin. These results suggest that P. grandiflorus flower extract may be useful as a natural cosmetic pigment.
Journal of Physiology & Pathology in Korean Medicine
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v.19
no.2
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pp.481-489
/
2005
Lonicerae Flos has antibacterial effects against Staphylococcus aureus, streptococci, pneumococci, Bacillus dysenterii, Salmonella typhi, and paratyphoid. It is an antiviral agent. The herb has a cytoprotective effect against $CCl_{4}-induced$ hepatic injury. It has antilipemic action, interfering with lipid absorption from the gut. Nowadays this herb is used mainly in the treatment of upper respiratory infections, such as tonsillitis and acute laryngitis. It is also used in the treatment of skin suppurations, such as carbuncles, and to treat viral conjunctivitis, influenza, pneumonia, and mastitis. Lonicerae Flos is dried flower buds of Lonicera japonica, L. hypoglauca, L. confusa, or L. dasystyla. But, for the most part, we use whole plant of Lonicera japonica, as a flower bud of it. And, little is known of the original copy of effects of whole plant, except for the 'Bon-Cho-Gang-Mok', which is written the effects of flower of Lonicera japonica are equal to effects of leaves and branch of it. The present study was conducted to evaluate the effect of flower and whole plant of Lonicera japonica on the regulatory mechanism of cytokines, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) for the immunological activities in Raw 264.7 cells. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, flower and whole plant of Lonicera japonica water extracts inhibited nitric oxide production in a dose-dependent manner and abrogated iNOS and COX-2. Flower and whole plant of Lonicera japonica water extract did not affect on cell viability. To investigate the mechanism by which flower and whole plant of Lonicera japonica water extract inhibits iNOS and COX-2 gene expression, we examined the on phosphorylation of inhibitor ${\kappa}B{\alpha}$ and assessed production of $TNF-{\alpha}$, $interleukin-1{\beta}$$(IL-1{\beta})$ and interleukin-6 (IL-6). Results provided evidence that flower and whole plant of Lonicera japonica inhibited the production of $IL-1{\beta}$, IL-6 and activated the phosphorylation of inhibitor ${\kappa}B{\alpha}$ in Raw 264.7 cells activated with LPS. These findings suggest that flower and whole plant of Lonicera japonica can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections, respectively.
Jeong Ah Lee;Hye Min Seol;Seong Un Jeong;Jae Hyeon Yoon;Jeong Soo Bae;Tae Hee Kim;Hyeong Soo Kim
Journal of Life Science
/
v.34
no.8
/
pp.558-566
/
2024
The potential therapeutic effects of Kaempferia Parviflora ethanol extract (KPE) on osteoarthritis were investigated using the human chondrocyte cell line (CHON-001) to explore its application in functional foods. The CHON-001 cells were pre-treated with either 1 ㎍/ml or 5 ㎍/ml of KPE before exposure to 10 ng/ml of IL-1β to induce osteoarthritis. Results showed that KPE treatment significantly suppressed IL-1β-induced TNF-α production by 66% and 50% at concentrations of 1 ㎍/ml and 5 ㎍/ml KPE, respectively. In addition, COX-2 protein expression was reduced by 26% and 34% compared to control levels. The preservation of chondrocyte-specific matrix proteins, aggrecan, and collagen type II, was notable, with aggrecan and mRNA levels maintained by 5% and 8%, and collagen II levels preserved by 62% and 47% at the same KPE concentrations. This preservation is likely due to the reduced expression of MMP1 and MMP13, enzymes responsible for matrix protein degradation. Overall, the current results suggest that KPE may protect chondrocytes from IL-1β-induced osteoarthritis by suppressing TNF-α production and COX-2 expression while preserving critical matrix proteins like aggrecan and collagen II by suppressing the expressions of their degrading enzymes (MMP-1 and MMP-13). Therefore, KPE holds promise as a candidate for developing functional foods aimed at reducing osteoarthritis.
Background: Collagen-induced arthritis (CIA) in mice is animal model of autoimmune disease known as rheumatic arthritis in human. We investigated CII-specific CD4+ T cell receptor usage in CIA mice. Methods: In CIA model, draining lymph node (dLN) CD4+ T cells and splenocytes at $3^{rd},\;5^{th},\;8^{th}$ week, we investigated CII-specific T cell proliferation, production of IL-17, IFN-${\gamma}$, TNF-${\alpha}$, IL-4 and IL-10. And we also performed anti-CII IgG Ab measurements in serum level, TCRV ${\beta}$ usage and T cell clonality with RT-PCR-SSCP analysis. Also, we performed proliferative response against CII when CII-specific T cell subset is deleted. Results: CIA mice showed more increase in the serum level of anti-CII IgG than normal mice after induction of arthritis. And the level of anti-CII IgG2a in CIA mice was increased after $3^{rd}$ week after primary immunization, while anti-CII IgG1 was decreased. Draining LN CD4+ T cells have proliferated against CII stimulation at $3^{rd}$ week after $1^{st}$immunization. CD4+T cells derived from dLN of CIA mice produced proinflammatory cytokine IFN-${\gamma}$, IL-17 etc. Draining LN CD4 T cells of CIA presented higher proportion of CD4+V ${\beta}3$+subset compared to those of normal mice at $3^{rd}$ week after $1^{st}$ immunization, and they were increased in proportion by CII stimulation. Draining LN CD4+ T cells without TCRV ${\beta}3+/V{\beta}8.1/8.2+/V{\beta}$10b+cells were not responsive against CII stimulation. But, CII-reactive response of TCRV ${\beta}3-/V{\beta}8.1/8.2-/V{\beta}$10b- T cells was recovered when $V{\beta}3+$ T cells were added in culture. Conclusion: Our results indicate that CD4+$V{\beta}3+$ T cells are selectively expanded in dLN of CIA mice, and their recovery upon CII re-stimulation in vitro, as well as the production Th1-type cytokines, may play pivotal role in CIA pathogenesis.
The anti-inflammatory effect of Sargassum patens C. Agardh ethanol extract (SPEE) was examined based on the lipopolysaccharide (LPS)-induced inflammatory response in this study. SPEE treatment was not cytotoxic to macrophages compared to the control. The production of NO was suppressed by SPEE by approximately 28% at $100{\mu}g/ml$, and levels of interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$ decreased in a dose-dependent manner. In addition, the expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ was suppressed by SPEE treatment. In vivo, croton oil-induced mouse ear edema was attenuated by SPEE and the infiltration of mast cells into the tissue decreased. Based on these results, SPEE inhibits the release of LPS-induced pro-inflammatory cytokines and mediators, suggesting that SPEE is a potential agent for anti-inflammatory therapies.
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