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Inhibition of Inflammation by Kyeongok-go with Black ginseng in LPS-induced RAW 264.7 Macrophages (LPS로 염증유도된 RAW 264.74 세포에 대한 흑삼 첨가 경옥고의 항염증 효과)

  • Park, MyungJae;Kim, Jeong-Soo;Lee, AhReum;Roh, Seong-Soo;Kwon, OJun;Seo, Young-Bae
    • The Korea Journal of Herbology
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    • v.32 no.3
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    • pp.19-27
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    • 2017
  • Objectives : The aim of this study was to examine effect of anti-oxidant and anti-inflammation activity of the Kyeongok-go with various processing methods that was manufactured by heating mantle. Methods : Commercial Kyeongok-go (K0) was purchased and Kyeongok-go with ginseng (K1), Kyeongok-go with black ginseng (BK), ginseng fermentation Kyeongok-go (KF), black ginseng fermentation Kyeongok-go (BKF) were manufactured by heating mantle. To examine anti-oxidant effect, DPPH radical and production of NO and ROS in RAW 264.74 cell were used. Furthermore, to determined anti-inflammation effect, measured pro-inflammatory mRNA such as NOS-II, COX-2, $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ in RAW 264.74 cell treated with K0, K1, KF, BK, and BKF. Result : K1 scavenged DPPH radical effectively than K0. The most DPPH radical scavenging activity was BKF. In the RAW 264.74 cells stimulated with LPS, NO and ROS production were measured. As a results, K1 was decreased NO, ROS production compared with K0, and BKF was reduced similarly to cyclosporine A (positive control). Expression of pro-inflammatory mRNA such as NOS-II, COX-2, $IL-1{\beta}$, IL-6 showed a significant decrease in BK or BKF. But, there was no significant in expression of $TNF-{\alpha}$ in all extract treatmetn groups. Conclusions : According to the above results, it is considered that Kyeongok-go with fermented black ginseng (BKF) manufactured by heating mantle is effective material that have anti-inflammation and anti-oxidant activities. Our finding indicate that BKF may be an effective agent for anti-inflammation through anti-oxidant effect.

Odontogenic Ameloblast-Associated Protein (Odam) Plays Crucial Roles in Osteoclast Differentiation via Control of Actin Ring Formation

  • Lee, Hye-Kyung;Park, Joo-Cheol
    • Journal of Korean Dental Science
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    • v.8 no.2
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    • pp.74-81
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    • 2015
  • Purpose: In osteoclast differentiation, actin-rich membrane protrusions play a crucial role in cell adhesion. Odontogenic ameloblast-associated protein (Odam) contributes to cell adhesion by inducing actin rearrangement. Odam-mediated RhoA activity may play a significant role in multinucleation of osteoclasts. However, the precise function of Odam in osteoclast cell adhesion and differentiation remains largely unknown. Here, we identify a critical role for Odam in inducing osteoclast adhesion and differentiation. Materials and Methods: The expression of Odam in osteoclasts was evaluated by immunohistochemistry. Primary mouse bone marrow and RAW264.7 cells were used to test the cell adhesion and actin ring formation induced by Odam. Result: Odam was expressed in osteoclasts around alveolar bone. Odam transfection induced actin filament rearrangement and cell adhesion compared with the control or collagen groups. Overexpression of Odam promoted actin stress fiber remodeling and cell adhesion, resulting in increased osteoclast fusion. Conclusion: These results suggest that Odam expression in primary mouse osteoclasts and RAW264.7 cells promotes their adhesion, resulting in the induction of osteoclast differentiation.

Inhibitory Effects of Ginsenoside Rb1,Rg3, and Panax ginseng Head Butanol Fraction on Inflammatory Mediators from LPS-Stimulated RAW 264.7 Cells

  • Lee, Je-Hyuk;Jeong, Choon-Sik
    • Biomolecules & Therapeutics
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    • v.16 no.3
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    • pp.277-285
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    • 2008
  • Panax ginseng C.A. Mayer (Araliaceae, P. ginseng) has been used for the enhancement of vascular and immune functions in Korea and Japan for a long time. Ginsenoside $Rb_1$ and $Rg_3$ isolated from P. ginseng head-part butanolic extract (PGHB) were investigated for anti-inflammatory activity. Ginsenosides and PGHB did not affect the cell viability within $0\;-\;100\;{\mu}g/ml$ concentration to RAW 264.7 murine macrophage cells. Ginsenosides and PGHB inhibited partly lipopolysaccharide (LPS)-induced nitrite production in a dose-dependent manner. The ginsenosides and PGHB showed partially chemical nitric oxide (NO) quenching (maximum 40%) in the cell-free system. Also, ginsenoside $Rb_1$ and $Rg_3$ inhibited markedly approximately 74 and 54% of inducible nitric oxide synthase (iNOS) mRNA transcription from LPS-induced RAW 264.7 cells. Taken together, the inhibitory effect of ginsenosides and PGHB on NO production did not occur as a result of cell viability, but was caused by both the chemical NO quenching and the regulation of iNOS. Additionally, the ginsenoside $Rb_1$ and PGHB inhibited prostaglandin $E_2$ ($PGE_2$) synthesis in a concentration-dependent manner, showed approximately 70-98% inhibition at $100\;{\mu}g/ml$ concentration. And the treatment with ginsenosides and PGHB attenuated partially LPS-upregulated cyclooxygenase-2 (COX-2) gene transcription. Ginsenoside $Rg_3$ suppressed LPS-stimulated interleukin-6 (IL-6) level to the basal in RAW 264.7 cells. From these results, ginsenoside $Rb_1,\;Rg_3$, and PGHB may be useful for the relief and retardation of immunological inflammatory responses and its action may occur through the reduction of inflammatory mediators, including NO, $PGE_2$, and IL-6 production.

Verification of the Physiological Activity of Geranium thunbergii Extract and Anti-inflammatory Activity in Raw 264.7 Cells (현지초(Geranium thunbergii) 추출물의 생리활성 및 Raw 264.7 cells에서의 항염활성 검증)

  • Seung-Mi Park;Min-Jeong Oh;Jin-Young Lee
    • Journal of Life Science
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    • v.34 no.1
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    • pp.28-36
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    • 2024
  • We evaluated the efficacy of Geranium thunbergii (GT), which has so far been understudied as a cosmetic material, and conducted anti-inflammatory-related activity studies. We measured the electron donation ability and ABTS+ radical scavenging ability to confirm the antioxidant ability of GT and found values of 91% and 94% at a concentration of 50 ㎍/ml, respectively, confirming that GT had excellent antioxidant ability. Tyrosinase inhibitory activity was measured to evaluate whitening activity, and it was found that inhibitory activity was 24.8% at the highest concentration of 1,000 ㎍/ml. Elastase and collagenase inhibitory activity were measured to determine the wrinkle improvement activity of the GT; 30.6% and 90% inhibitory activity were shown at the highest concentration of 1,000 ㎍/ml, respectively. Excellent inhibitory activity was confirmed through the measurement of collagenase inhibitory activity. Before the cell experiments were conducted, the survival rate of the macrophages Raw 264.7 according to GT treatment was determined based on the MTT assay, and the cell survival rate was greater than 83.6% at a concentration of 100 ㎍/ml. Subsequent cell-related experiments were conducted at concentrations of 100 ㎍/ml or less. The NO production inhibitory activity according to the GT treatment by NO assay was measured, and a 74.9% inhibitory rate was confirmed at a concentration of 100 ㎍/ml. Western blotting was performed to determine protein expression inhibition, and both COX-2 and iNOS factors were concentration-dependently inhibited in GT. Based on these results, GT is considered to have potential as an anti-inflammatory functional cosmetic material.

Effects of Ethanol Extract from Lathyrus palustris on Anti-inflammation Response of RAW 264.7 Cell (RAW 264.7 대식세포 염증반응에 대한 털연리초 에탄올 추출물의 항염증 효과)

  • Nam, Jung Hwan
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.287-292
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    • 2020
  • Lathyrus palustris often used as a treatment for inflammation of the kidneys in Korean traditional medication. Generally, drugs for arthritis have anti-inflammatory and antinociceptive properties. However, the validity of the anti-inflammatory effect has not been scientifically investigated so far. Therefore, the purpose of the research was to investigate the latent anti-inflammatory ability of L. palustris using the ethanol extract. To evaluate the anti-inflammatory activities, we examined the inflammatory arbitrators such as a nitric oxide (NO) and prostaglandin E2 (PGE2) on RAW 264.7 cells. Our results indicated that ethanol extract significantly inhibited the lipopolysaccharide E (LPS) derived PGE2 production in RAW 264.7 cell. The inhibitory activity of ethanol extract for PGE2 tests with inhibition ratio showed in 40 ㎍/mL. Overall, PGE2 tests had a higher inhibitory effect on inflammation than NO tests. This result anticipated that the ethanol extract from L. palustris is a good candidate for developing the origin of anti-inflammatory agents.

Anti-inflammatory Effect of Unripe Fruit of Citrus grandis Osbeck in RAW 264.7 and HaCaT Cells (RAW 264.7 및 HaCaT Cell에서 당유자 미숙과의 염증억제 효과)

  • Lee, Hye-Ja;Kang, Gyeoung-Jin;Yoon, Weon-Jong;Kang, Hee-Kyoung;Kim, Young-Suk;Kim, So-Mi;Yoo, Eun-Sook
    • Korean Journal of Pharmacognosy
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    • v.37 no.2 s.145
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    • pp.74-80
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    • 2006
  • We investigated the anti-inflammatory activities of unripe fruit of Citrus grandis Osbeck growing at Jeju Island, through the evaluation of their inhibitory effect on the production of inflammatory markers (IL-6, iNOS, COX, TARC and MDC) in RAW264.7 murine macrophage cells and HaCaT human keratinocyte cells. Among the sequential solvent fractions obtained from crude extract, hexane and chloroform $(CHCI_3)$ fractions showed potential inhibitory activity on the mRNA expressions of IL-6, iNOS and COX-2 at the concentration of $100\;{\mu}g/ml$ in RAW264.7 cells. Also, EtOAc fraction showed inhibitory activity on the thymus and activation-regulated chemokine (TARC)/CCL17 and macrophage-derived chemokine (MDC)/CCL22 at the concentration of $50\;{\mu}g/ml$ in HaCaT cells. These results suggest that the unripe fruit of C. grandis may have anti-inflammatory activity through the suppression of inflammatory markers (IL-6, iNOS, COX, TARC and MDC).

Effects of Fermented Soybean upon Anti-inflammation and Intestinal Mucous Membrane Permeability (청국장의 항염증 및 장점막 투과성 개선 효과)

  • Kim, Hyung-Gu;Lee, Myeong-Jong;Kim, Ho-Jun;Kim, Ki-Cheol;Bose, Shambhunath
    • Journal of Korean Medicine for Obesity Research
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    • v.12 no.1
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    • pp.33-47
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    • 2012
  • Objectives This study was designed to investigate the effects of fermented soybean upon anti-inflammation, cytotoxicity, antioxidant and intestinal mucous membrane permeability by measuring the cell viability, NO (nitric oxide) production, DPPH, Polyphenol, HRP and TEER in cells like Raw 264.7 and HCT 116 using fermented soybean. Methods Raw 264.7 cell and HCT 166 cell were used in this study. And fermented soybean powders were used for the experimental group and soybean powders for the control group. There was inflammation response upon using lipopolysaccharide(LPS). Fermented soybean powders and soybean powders were in a respectively different dose added to the cells with LPS. MTT assay, NO, DPPH and Polyphenol measurement, TEER, HRP were conducted for each cell. The results of this study were presented in mean and standard deviation. Results 1. In Raw 254.7 cells added with $100{\mu}l/ml$ unfermented soybean powders, 104.95% higher than 62.59% was measured. In Raw 254.7 cells added with $100{\mu}l/ml$ fermented soybean powders, there was 74.90% measured higher than 62.59%, which was a significant result. 2. By a gradual increase of unfermented soybean powders like $0.1{\mu}l/ml$, $1.0{\mu}l/ml$, $10{\mu}l/ml$, $100{\mu}l/ml$, the measured NO were also gradually decreased $53.12{\mu}M$, $47.57{\mu}M$, $37.02{\mu}M$, $28.16{\mu}M$. In case of cells added with fermented soybean powders, $43.95{\mu}M$ NO was measured in $0.1{\mu}l/ml$ which is significant, and in other cases, mostly measured over$ 56.72{\mu}M$. 3. It was inferred that fermented soybean powders have anti-inflammatory effects of maintaining intestinal mucous membrane permeability because the measured values of cells in both groups were all higher than $133.62{\Omega}$ measured of cells added with only LPS. And measured values of cells in both groups were all lower than 2.26 measured of cells added with only LPS. 4. In case of experiment DPPH and polyphenol measurement, fermented group was all higher than unfermented group. Conclusion From the results of conducting MTT assay, NO measurement, and TEER, HRP by using cells Raw 264.7 and HCT-116, even though there was no significance in the correlation between cytotoxicity, anti-inflammatory effects, both unfermented soybean powders and fermented soybean powders were shown to have intestinal mucous membrane permeability improvement effects. This effects could be applicable for autoimmune diseases, chronic inflammatory diseases and so additional studies are expected in the future. From the results of conducting DPPH, Polyphenol measurement, Fermented soybean may be useful as potential antioxidant.

The Effect of Bacillus-Fermented Scutellariae Radix Acupuncture Solution on Interleukin Production in Mouse Macrophage Stimulated by Lipopolysaccharide (바실러스균 발효황금약침액이 Lipopolysaccharide로 활성화된 마우스 대식세포의 인터루킨 생성에 미치는 영향)

  • Park, Wan-Su
    • Korean Journal of Acupuncture
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    • v.27 no.2
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    • pp.95-105
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    • 2010
  • Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on interleukin(IL) production in mouse macrophage stimulatedby lipopolysaccaride(LPS). Methods : Productions of interleukins were measured y High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$(multi-analyte profiling beads) technology. To begin with, cell culture supernatant was obtained after treatment with LPS(1 ${\mu}g/mL$) and SB for 24 hour. Then, it was incubated with the antibody-conj${\mu}g$ated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. After incubating for 30 minutes, Strepavidin-conjugated Phycoerythrin(SAPE) was then added. Incubating for another 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Results : The results of the experiment are as follows. SB significantly inhibited the LPS-induced production of IL-3($9.15{\pm}0.35$ pg/mL) by $6.92{\pm}0.05,\;7.21{\pm}0.11,\;6.96{\pm}0.33,\;and\;7.45{\pm}0.74$ pg/mL at the concentration of 25, 50, 100, and 200 ${\mu}g/mL$ in mouse macrophage RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced production of IL-5($7.30{\pm}0.48$ pg/mL) by $6.50{\pm}0.29,\;6.30{\pm}0.25,\;6.30{\pm}0.25,\;and\;5.80{\pm}0.25$ pg/mL at the concentration of 25, 50 100, and 200 ${\mg}g/mL$ in RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced productiion of IL-9($17.26{\pm}0.19$ pg/mL) by $15.01{\pm}0.43$ pg/mL at the concentration of 25 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced productioh of IL-13($187.80{\pm}2.90$ pg/mL) by $152.80{\pm}4.25,\;172.80{\pm}3.97,\;162.10{\pm}6.67,\;and\;165.30{\pm}11.80$ pg/mL at the concentration fo 25, 50, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-17($18.30{\pm}0.95$ pg/mL) by $13.30{\pm}1.25,\;13.80{\pm}1.11,\;13.30{\pm}0.75,\;and\;14.00{\pm}1.08$ pg/mL at the concentration of 25, 50 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-23($43.90{\pm}0.83$ pg/mL by $39.50{\pm}1.26,\;38.00{\pm}1.78,\;and\;39.60{\pm}2.49$ pg/mL at the concentration of 25, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). Conclusions : These results suggest that SB has anti-inflammatory activity related with its inhibition of IL-3, IL-5, IL-13, IL-17, and IL-23 production in macrophages.

Enhancement of Anti-inflammatory Activity of Lactobacillus plantarum Fermented by Achyranthes japonica on Extraction Solvents (추출 용매에 따른 Lactobacillus plantarum 발효 우슬의 항염증 효과 증진)

  • Jo, Eun Sol;Woo, Young Min;Kim, Ok Ju;Jo, Min Young;Ahn, Mee Young;Lee, Jae-Hwa;Ha, Jong-Myung;Kim, Andre
    • Applied Chemistry for Engineering
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    • v.30 no.2
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    • pp.145-150
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    • 2019
  • In this study, we used extracts obtained from five different solvents (water, ethanol, hexane, ethyl acetate, butanol) of Achyranthes japonica (AJ) and also AJ fermented with Lactobacillus plantarum (LP) to confirm effects on the anti-inflammatory activity in RAW264.7 cells. Experiments of measuring nitric oxide (NO) and cytokine production were performed in lipopolysaccharide (LPS)-induced RAW264.7 cells, and the expression of both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was observed by a western blot method. The cytotoxicity of RAW264.7 was confirmed by the cell counting kit (CCK) assay at a concentration of $100{\mu}g/mL$, which has no toxicity. As a result of the inhibition of NO production, the inhibition rate of AJ-LP extracted with ethanol samples was about 74% higher than that of using the control group. Interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and Interleukin-$1{\beta}$ (IL-$1{\beta}$), which are inflammatory cytokines, also showed an excellent efficacy with inhibition rates of about 57, 70, and 74%, respectively. Comparing to the results of COX-2 and iNOS expression in the AJ group, the inhibition rate of 20-hydroxyecdysone was the highest than others. On the other hand, the COX-2 expression level of AJ-LP group decreased about 16% compared to that of the control group, and the iNOS expression level was also decreased about 7%. These results suggest that the extract of AJ fermented from L. plantarum can be used as an anti-inflammatory natural material.

Evaluation of the Cosmeceutical Activity of Apocynum lancifolium Russanov Extracts (개정향풀 추출물의 향장학적 활성 평가)

  • Park, Ji Hyo;Lee, Ji-An
    • Journal of Convergence for Information Technology
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    • v.11 no.1
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    • pp.236-243
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    • 2021
  • In this study, the functional cosmetic activity of hot-water (AW) and methanol(AM) extracts from Apocynum lancifolium Russanov are examined. In the DPPH antioxidant activity test, the AW extract showed the highest inhibition rate of 90.5% (IC50 37.717 ± 8.209 ㎍/mL) and the antioxidative activity of ABTS showed a high activity in the AW extract with an IC50 of 185.244 ± 12.602 ㎍/mL and 96.2%, respectively. In RAW264.7 macrophages, the two extracts significantly suppressed the LPS-induced nitric oxide (NO), TNF-α, IL-6 production and iNOS expression level. The MTT assay measured by the cell survival rate showed that AW and AM extracts have no toxicity. The astringent activity of the AW extract exhibited the highest astringent activity with 74.336±2.487 mg/mL. Therefore, these results suggest that A. lancifolium extracts have a health promotion potential which can be further developed as food additive, natural products for cosmetic purpose.