• Korean Journal of Acupuncture
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• v.36 no.2
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• pp.115-126
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• 2019

Objectives : Osteoporosis is a condition characterized by low bone mass and increased bone fragility. It has become a major problem of senior citizens. The purpose of this study is to experiment the effect of water extract of Forsythiae Fructus (wFF) on osteoclast differentiation; and the other purpose is to examine the effect of wFF on osteoporosis in ovariectomized rat. Methods : To investigate the effect of wFF on osteoclast differentiation and activity, RAW 264.7 cells were used. The number of TRAP positive cell, TRAP activity, pit area, mRNA expression of makers (RANK, TRAP, CA II, CTK, MMP-9, NFATc1, c-Fos), protein expression of makers (NFATc1, c-Fos) were investigated. For in vivo study, 40 female Sprague-Dawley (SD) rats were induced osteoporosis by ovariectomy (OVX) and then tested for anti-osteoporosis effect by administration of wFF. Results : wFF suppressed osteoclatogenesis, TRAP activity and pit area formation. Moreover, wFF decreased the expression of master differentiation factors (NFATc1, c-Fos) and also reduced the osteoclastogenesis-related markers (TRAP, CA II, CTK, MMP-9). These suggest that wFF inhibit osteoclasts differentiation and bone resorption. In the OVX rat model, wFF inhibited decreasing of BMD and trabecular area. Conclusions : Forsythiae Fructus should be effective for osteoporosis prevention and treatment.

• Journal of Ginseng Research
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• v.45 no.1
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• pp.75-85
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• 2021

Background: Invasive infections due to foodborne pathogens, including Salmonella enterica serovar Typhimurium, are prevalent and life-threatening. This study aimed to evaluate the effects of ginsenoside Rg3 (Rg3) on the adhesion, invasion, and intracellular survival of S. Typhimurium. Methods: The impacts of Rg3 on bacterial growth and host cell viability were determined using the time kill and the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays, respectively. Gentamicin assay and confocal microscopic examination were undertaken to determine the effects of Rg3 on the adhesive and invasive abilities of S. Typhimurium to Caco-2 and RAW 264.7 cells. Quantitative reverse transcription polymerase chain reaction was performed to assess the expression of genes correlated with the adhesion, invasion, and virulence of S. Typhimurium. Results: Subinhibitory concentrations of Rg3 significantly reduced (p < 0.05) the adhesion, invasion, and intracellular survival of S. Typhimurium. Rg3 considerably reduced (p < 0.05) the bacterial motility as well as the release of nitrite from infected macrophages in a concentration-dependent manner. The expression of genes related to the adhesion, invasion, quorum sensing, and virulence of S. Typhimurium including cheY, hilA, OmpD, PrgK, rsgE, SdiA, and SipB was significantly reduced after Rg3 treatment. Besides, the compound downregulated rac-1 and Cdc-42 that are essential for actin remodeling and membrane ruffling, thereby facilitating Salmonella entry into host cells. This report is the first to describe the effects of Rg3 on "trigger" entry mechanism and intracellular survival S. Typhimurium. Conclusion: Rg3 could be considered as a supplement agent to prevent S. Typhimurium infection.

• Microbiology and Biotechnology Letters
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• v.50 no.2
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• pp.310-318
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• 2022

The fruits of Paulownia tomentosa (Thunb.) (PT) Steud. have been reported to exert a variety of biological activities. A previous study confirmed that compounds isolated from PT fruits (PTF) exerted anti-inflammatory effects on TNF-α-stimulated airway epithelial cells. However, there is no report on the protective effects of PTF on acute lung injury (ALI). Here, we examined the ameliorative effects of PTF in an experimental animal model of lipopolysaccharide (LPS)-induced ALI. In ALI mice, increased levels of inflammatory cell influx were confirmed in the lungs of mice, and an increase of microphage numbers, TNF-α, IL-6 and MCP-1 production and protein content were detected in mouse bronchoalveolar lavage fluid. However, these increases were significantly reversed with PTF pretreatment. In addition, PTF inhibited the increased expression of iNOS and COX-2 in the lungs of ALI mice. Furthermore, the upregulation of MAPK and NF-κB activation was decreased in the lungs of ALI mice by PTF. In the in vitro experiment, PTF pretreatment exerted an anti-inflammatory effect by inhibiting the secretion of nitric oxide, TNF-α and IL-6 in LPS-stimulated RAW264.7 macrophages. Collectively, these results indicated that PTF has ameliorative effects on airway inflammation in an experimental animal model of ALI.

• Parasites, Hosts and Diseases
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• v.60 no.2
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• pp.117-126
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• 2022

Cystatin, a cysteine protease inhibitor found in many parasites, plays important roles in immune evasion. This study analyzed the molecular characteristics of a cystatin from Fasciola hepatica (FhCystatin) and expressed recombinant FhCystatin (rFhcystatin) to investigate the immune modulatory effects on lipopolysaccharide-induced proliferation, migration, cytokine secretion, nitric oxide (NO) production, and apoptosis in mouse macrophages. The FhCystatin gene encoded 116 amino acids and contained a conserved cystatin-like domain. rFhCystatin significantly inhibited the activity of cathepsin B. rFhCystatin bound to the surface of mouse RAW264.7 cells, significantly inhibited cell proliferation and promoted apoptosis. Moreover, rFhCystatin inhibited the expression of cellular nitric oxide, interleukin-6, and tumor necrosis factor-α, and promoted the expression of transforming growth factor-β and interleukin-10. These results showed that FhCystatin played an important role in regulating the activity of mouse macrophages. Our findings provide new insights into mechanisms underlying the immune evasion and contribute to the exploration of potential targets for the development of new drug to control F. hepatica infection.

• Journal of Microbiology and Biotechnology
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• v.31 no.6
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• pp.855-866
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• 2021

The effects of various carbon sources on mycelial growth and polysaccharide synthesis of the medicinal fungus Inonotus obliquus in liquid fermentation were investigated. After 12-d fermentation, mycelial biomass, polysaccharide yield, and polysaccharide content were significantly higher in Glc+Lac group (glucose and lactose used as combined carbon source) than in other groups. Crude polysaccharides (CIOPs) and the derivative neutral polysaccharides (NIOPs) were obtained from mycelia fermented using Glc, fructose (Fru), Lac, or Glc+Lac as carbon source. Molecular weights of four NIOPs (termed as NIOPG, NIOPF, NIOPL, and NIOPGL) were respectively 780.90, 1105.00, 25.32, and 10.28 kDa. Monosaccharide composition analyses revealed that NIOPs were composed of Glc, Man, and Gal at different molar ratios. The NIOPs were classified as α-type heteropolysaccharides with 1→2, 1→3, 1→4, 1→6 linkages in differing proportions. In in vitro cell proliferation assays, viability of RAW264.7 macrophages was more strongly enhanced by NIOPL or NIOPGL than by NIOPG or NIOPF, and proliferation of HeLa or S180 tumor cells was more strongly inhibited by NIOPG or NIOPGL than by NIOPF or NIOPL, indicating that immune-enhancing and anti-tumor activities of NIOPs were substantially affected by carbon source. qRT-PCR analysis revealed that expression levels of phosphoglucose isomerase (PGI) and UDP-Glc 4-epimerase (UGE), two key genes involved in polysaccharide synthesis, varied depending on carbon source. Our findings, taken together, clearly demonstrate that carbon source plays an essential role in determining structure and activities of I. obliquus polysaccharides by regulating expression of key genes in polysaccharide biosynthetic pathway.

• The Korea Journal of Herbology
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• v.28 no.5
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• pp.45-52
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• 2013

Objectives : This study aims at examining the immuno-modulating activity in the fermentative extract of the root of Scutellaria baicalensis Georgi (Scutellariae Radix) on the production of inflammatory mediator in LPS-stimulated RAW264.7 mouse macrophages. Method : Measurements were done for the influences on the cell viability, generation of hydrogen peroxide in cells and nitric oxide (NO) generation using the macrophage of mouse with the specimen SBS as the fermentative extract of Scutellariae Radix (SBS) with Saccharomyces cerevisiae STV89. Result : As a result of carrying out MTT assay to check the cellular toxicity of the fermentative extract of Scutellariae Radix, any excessive toxicity to the macrophage did not occur from treatments by concentration for SBS. SBS increased the generation of hydrogen peroxide in the macrophage. SBS suppressed the NO generated in macrophages and SBS concentration higher than $25{\mu}g/mL$ significantly suppressed the increased NO generated in LPS-stimulated macrophages. SBS concentration higher than $25{\mu}g/mL$ significantly suppressed the generation of IL-6, IL-10, IL-12p40 and MCP-1 in LPS-stimulated macrophages. Conclusion : Our findings indicate that SBS has an immuno-modulating activity in macrophage activation through suppressing the generation of inflammatory substances, NO, IL-6, IL-10, IL-12p40 and MCP-1.

• The Korea Journal of Herbology
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• v.34 no.6
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• pp.117-124
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• 2019

Objective : Broccoli is edible green plant that has a wide variety of health benefits including cancer prevention and cholesterol reduction. However, leaves of broccoli are not eaten and are mostly left as waste. This study was conducted to evaluate the effects of the broccoli leaf extract (BLE) on prostaglandin E₂ (PGE₂) production related to nuclear factor kappa B (NF-κB) signaling in lipopolysaccharide (LPS)-activated macrophages. Methods : BLE was prepared by extracting dried leaf with ethanol. Cell viability was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. PGE₂ and inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA). Expression level of each protein was monitored by Western blot analysis. Results : In LPS-activated Raw264.7 cells, PGE₂ release into culture medium was dramatically enhanced compared to control cells. However, increased PGE₂ was attenuated dose-dependently by treatment with BLE. Inhibition of PGE₂ production by BLE was due to the suppression of cyclooxygenase-2 (COX-2) expression determined by Western blot analysis. BLE also inhibited the production of inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α). Inhibition at PGE₂ and cytokine was mediated from inhibition of nuclear translocation of NF-κB due to the repression of inhibitory kappa B alpha (IκBα) phosphorylation and degradation. Conclusion : This study showed that BLE exerted inhibitory activities against PGE₂, which is critical for the initiation and resolution of inflammatory responses, and that inhibition of PGE₂ was mediated by suppression of NF-κB signaling. These results suggest that the waste broccoli leaves could be used for controlling inflammation.

• The Korea Journal of Herbology
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• v.32 no.2
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• pp.49-56
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• 2017

Objectives : This study investigated the anti-oxidant activities and improving effect of Phellinus linteus and Glycyrrhiza uralensis Extract (PGE) on Atopic Dermatitis. Methods : 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radical, 2,2′-azino-bis 3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical, Hydrogen peroxides scavenging activities and Superoxide dismutase (SOD)-like activities were used for the measurement of anti-oxidant ability. Cytotoxicity of PGE in Raw 264.7 cell was evaluated by MTT assay. To evaluate the anti-atopic dermatitis effect of PGE, a total of 33 patients with atopic dermatitis were observed trans epidermal water loss, skin moisture content, modified SCORAD index of atopic dermatitis and pruritic degree after applying the PGE for 4 weeks. Results : PGE scavenged DPPH ($IC_{50}=25ppm$) effectively, ABTS and Hydrogenperoxides scavenged similar to BHA. As for the SOD-like activity, it had lower effect than ascorbic acid, but it comparable activities in 500ppm. There was no cytotoxicity at PGE at concentrations of 10,000ppm. In clinical research about PGE on patients with atopic dermatitis, skin condition was improved. After 4 weeks, the application of PGE increased skin moisture content from 19.43 to 31.22. Moreover, it reduced the skin temperature (from 32.5 to 31.9), skin pH (from 5.39 to 5.22), trans epidermal water loss (from 39.03 to 24.46) and pruritus score (from 6.07 to 3.87). In addition, the Modified SCORAD index decreased from 31.28 to 20.3. Conclusions : In conclusion, PGE possesses anti-oxidant and anti-atopic dermatitis activities, thus it could be potentially valuable as anti-atopic dermatitis material.

• Journal of Korean Medicine Rehabilitation
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• v.33 no.1
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• pp.1-12
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• 2023

Objectives This study was conducted to find out the antioxidant and inflammatory improvement effects of Wolbigachul-tang. Methods To evaluate the antioxidant and anti-inflammatory effects of wolbigachul-tang, the total polyphenol and flavonoid contents were measured. And radical scavenging function was measured. In addition reactive oxygen species, nitric oxide, cytokines productions were measured by treating wolbigachul-tang in the lipopolysaccharide-induced RAW264.7 cells. We also measured the gene expression of inducible nitric oxide synthase, cyclooxygenase-2, cytokines too. Results Radical scavenging experiments showed significant concentration-dependent scavenging capacity increase. The amount of reactive oxygen species, nitric oxide, cytokines production in the cell all showed a significant concentration-dependent decrease compared to the control group. The gene expression of inducible nitric oxide synthase, cyclooxygenase-2, and cytokines also showed a significant concentration-dependent decrease compared to the control group. Conclusions In conclusion, it is believed that wolbigachul-tang can be applied to various inflammatory diseases.

• The Korea Journal of Herbology
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• v.24 no.4
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• pp.39-47
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• 2009

Objectives : Hwangnyeonhaedok-tang (Huanglian Jiedu Tang; HHT) has been widely used for purging' 'fire' and lessening virulence of any pathogenic organism. However it has been rarely conducted to evaluate the immuno-biological activity. In this study, we evaluated anti-inflammatory effects of HHT in LPS-activated Raw264.7 cells. Methods : Cells were treated with $1\;{\mu}g/ml$ of LPS 1 h prior to the addition of HHT. Cell viability was measured by MTT assay. The production of NO was determined by reacting cultured medium with Griess reagent. PGE2 and proinflammatory cytokines were detected by ELISA. Expression of iNOS, COX-2, $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ were analyzed by immunoblot analysis. Results : All three doses of HHT (0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity during the entire experimental period. The levels of NO and PGE2 were dramatically augmented by LPS compared to control. However, HHT extract dose-dependently reduced these increases. Expression of iNOS and COX-2 protein were also decreased by treatment with HHT extract. Furthermore, HHT extract significantly reduced the nuclear translocation of NF-${\kappa}B$ which is critical in regulating inflammation through transcription of iNOS and COX-2. In addition, HHT extract reduced the elevated production of inflammatory cytokines including TNF-$\alpha$, IL-$1{\beta}$ and IL-6. Conclusions : The results in this study demonstrate that HHT extract exerts anti-inflammatory activities through the inhibition of NO, PGE2 and proinflammatory cytokines production via the suppression of NF-${\kappa}B$.