• Journal of Acupuncture Research
• /
• v.26 no.6
• /
• pp.1-9
• /
• 2009

Objectives : For evaluation of preventive and anti-diabetic activities of Cinnamomi ramulus(CR) herbal acupuncture on pancreatic islet damage in streptozotocin(STZ)-induced diabetic rat. Methods : CR herbal acupuncture was performed at Bisu($BL_{20}$) for 3 weeks subcutaneously starting1 week before STZ i.p. injection. SD rats were divided into four groups(n=10 for each group); 1) NC group, non-treated normal control group, 2) STZ group, STZ administered control group, 3) CR125 group, CR(125mg/kg) + STZ administered group, and 4) CR250 group, CR(250mg/kg) + STZ administered group. Results : Both of CR250 and CR125 groups showed increase in insulin secretion and decrease in the level of serum triglyceride and non-esterified fatty acid in a dose-dependent manner compared to the STZ group. Only CR250 group showed decrease in the levels of glucose and total cholesterol compared to the STZ group. CR herbal acupuncture prevents $\beta$-cell damage of pancreatic islet, showing round figure on the sections of the pancreas. In the pancreatic cells, expressions of iNOS, JNK-2, P-JNK-1/2 and ERK-1/2 were decreased compared to the STZ group. CR herbal acupuncture solution did not show any cytotoxicity by MTS assay and inhibited expressions of iNOS and COX-2 in the STZ-induced diabetic rats. Conclusions : Therefore, we suggest that CR herbal acupuncture may act as a prophylactic as well as a therapeutic modality for diabetes mellitus.

• The Korea Journal of Herbology
• /
• v.27 no.4
• /
• pp.99-107
• /
• 2012

Objectives : The aim of this study was to authenticate whether fractionated extract of Rumex japonicus HOUTT. (RJ) has anti-inflammatory effects in mouse macrophage, RAW264.7 cells. Methods : Roots of RJ were extracted by methanol for 48hours. The methanol that gained was filtered and freeze dried. The methanol extract was dissolved in water and dichloromethane (DCM). After that, two layers were separated. Ethyl acetate (EA) added to the water layer and separated again. All the layers were filtered and freeze dried and the extracts were tested. Cytotoxic activity of extracts on RAW 264.7 cells was measured using MTS assay. The nitric oxide (NO) production was measured and proinflammatory cytokines and $PGE_2$ were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), I ${\kappa}$-B-${\alpha}$ and nuclear NF-${\kappa}B$ p65 expression were detected by western blot. Results : Our results indicated that DCM and EA extracts of RJ inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 production in RAW 264.7 cells most effectively. DCM and EA extracts also had suppression effects of LPS-induced NF-${\kappa}B$ and MAPKs activation. Conclusions : This results demonstrate that fractionated extract of RJ has anti-inflammatory effects and among the fractioned extract, dichloromethane and ethyl acetate extract have best anti-inflammatory effects.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.23 no.3
• /
• pp.599-607
• /
• 2009

Prunellae Spica is the spike or whole plant of Prunella vulgaris Linne, which has been used for clearing heat from the liver, brightening the eyes and treating headache in traditional oriental medicines. This study was conducted to evaluate the inhibitory effects of the aqueous extract of Prunellae Spica (PSE; PS extract) on the production of NO and PGE2 in LPS-activated Raw 264.7 cells. Cell viability was determined by MTT assay, and all three doses of PS extract (0.03, 0.1 and 0.3 mg/ml) had no significant cytotoxicity during the entire experimental period. The cells were treated with 1 ${\mu}g/ml$ of LPS 1 h before adding PS extract, and increased NO and PGE2 production were detected in LPS-activated cells compared to control. However, these increases were dose-dependently attenuated by treatment with PS extract. The inhibition of NO by PS extract was due to the suppression of iNOS expression via inhibition of $NF{\kappa}B$ nuclear translocation and proteolytic degradation of $I{\kappa}B{\alpha}$. The decreased level of PGE2 was derived from inhibition of COX-2 activity, but expression of COX-2 protein was not affected by PS extract. Moreover, PS extract reduced the elevated production of IL-${\beta}$ and IL-6 by LPS. These results demonstrate that PS extract has inhibitory effects on the production of NO and PGE2 as a consequence of the reduction of proinflammatory cytokines, especially IL-${\beta}$ and IL-6 in LPS-activated Raw 264.7 cells.

• The Korea Journal of Herbology
• /
• v.23 no.3
• /
• pp.11-18
• /
• 2008

SunBangHwalMyungEum (SBH) has the effects of subduing swelling, resolving masses and alleviating pain in traditional oriental medicine. Recent studies showed that SunBangHwalMyungEum produced anti-cancer, anti-metastasis and immuno-modulatory effects. However there is lack of studies regarding the effects of SBH on the immunological activities. The present study was conducted to evaluate the effect of SBH on the regulatory mechanism of cytokines and nitric oxide (NO) in Raw 264.7 cells. Methods : After the treatment of SBH, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) was determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results : Results provided evidence that SBH inhibited the production of NO, iNOS, $interleukin-1{\beta}$ ($(IL-1{\beta})$), IL-6, and the activation of phospholylation of inhibitor ${\kappa}B{\alpha}$ in Raw 264.7 cells activated with lipopolysaccharide. Conclusions : These findings suggest that SBH can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• The Korean Journal of Physiology and Pharmacology
• /
• v.24 no.5
• /
• pp.395-402
• /
• 2020

This study has investigated the effect of a potent bioflavonoid, troxerutin, on diabetes-induced changes in pro-inflammatory mediators and expression of microRNA-146a and nuclear factor-kappa-B (NF-κB) signaling pathway in aortic tissue of type-I diabetic rats. Male Wistar rats were randomly divided into four groups (n = 6/each): healthy, healthy-troxerutin, diabetic, and diabetic-troxerutin. Diabetes was induced by streptozotocin injection (60 mg/kg; intraperitoneally) and lasted 10 weeks. Troxerutin (150 mg/kg/day) was administered orally for last month of experiment. Inflammatory cytokines IL-1β, IL-6, and TNF-α, as well as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM), cyclooxygenase-II (COX-II), and inducible-nitric oxide synthase (iNOS) were measured on aortic samples by enzyme-linked immunosorbent assay. Gene expressions for transcription factor NF-κB, interleukin-1 receptor-associated kinase-1 (IRAK-1), TNF receptor-associated factor-6 (TRAF-6), and microRNA-146a were determined using real-time polymerase chain reaction. Ten-week diabetes significantly increased mRNA levels of IRAK-1, TRAF-6, NF-κB, and protein levels of cytokines IL-1β, IL-6, TNF-α, adhesion molecules ICAM-1, VCAM, and iNOS, COX-II, and decreased expression of microRNA-146a as compared with healthy rats (p < 0.05 to p < 0.01). However, one month treatment of diabetic rats with troxerutin restored glucose and insulin levels, significantly decreased expression of inflammatory genes and pro-inflammatory mediators and increased microRNA level in comparison to diabetic group (p < 0.05 to p < 0.01). In healthy rats, troxerutin had significant reducing effect only on NF-κB, TNF-α and COX-II levels (p < 0.05). Beside slight improvement of hyperglycemia, troxerutin prevented the activation of NF-κB-dependent inflammatory signaling in the aorta of diabetic rats, and this response may be regulated by microRNA-146a.

• Journal of Life Science
• /
• v.32 no.7
• /
• pp.491-500
• /
• 2022

HK shiitake mushroom mycelium (HKSMM), containing 14% β-glucan, is a health functional food ingredient individually approved by the Korea Ministry of Food and Drug Safety for liver health. The anti-inflammatory effect of a 50% aqueous ethanol extract of HKSMM (designated HKSMM50) was studied in RAW 264.7 macrophage cells treated with lipopolysaccharide (LPS). An active hexose correlated compound (AHCC) was used as a positive control. LPS-activated RAW 264.7 cells were treated with HKSMM50 and AHCC (0, 20, 100, 500 ㎍/ml) and cultured for 24 hr. Inflammation-related elements in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA) kits, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins in the cells was analyzed by Western blotting. The HKSMM50 lowered iNOS and COX-2 protein expressions, and nuclear factor-kappa B (NF-κB), nitric oxide (NO) and prostaglandin E₂ (PGE₂) contents in a concentration-dependent manner as compared to LPS treatment. Similarly, the HKSMM50 lowered the content of pro-inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-6 (IL-6) contents and increased the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). The efficacy of the AHCC treatment was similar to that of the HKSSM50 treatments. These results indicate that HKSMM50 showed an anti-inflammatory effect in LPS-treated RAW 264.7 cells by down-regulation of NF-κB signaling and suggest that HKSMM could be used as a health functional food ingredient to help improve immune function.

• Journal of Korean Medicine Rehabilitation
• /
• v.27 no.1
• /
• pp.19-25
• /
• 2017

Objectives Eucommiae cortex is one of the frequently used herbs for musculoskeletal disorders, with well-known anti-inflammatory effect. Meanwhile, powdered form of herbal drugs is more advantageous than decoction form, in that storing and taking is more convenient. Thus, the authors aimed to investigate whether the drying method affects the index compound level and anti-inflammatory effect of eucommiae cortex water extract. Methods Eucommiae cortex was extracted in distilled water at $100^{\circ}C$ for 3 hours, filtered, and then evaporated under vacuum. One half of the sample was freeze-dried at $-80^{\circ}C$. Another half of the sample was added with dextrin and then spray-dried at $180^{\circ}C$. To assess the possible change in index compound content, pinoresinol diglucoside was selected as the index marker. The content level of the index compound in various extract sample was quantified through high performance liquid chromatography. In addition, iNOS assay in LPS-induced RAW 264.7 cells was adopted to determine the anti-inflammatory effect of various extract samples. Furthermore, MTT assay was performed to confirm that the result of iNOS assay was not due to cytotoxicity. Results There was no significance difference in index compound content between extract samples obtained through two different drying methods. Anti-inflammatory activity of extract samples were similar at the matching concentration, regardless of the drying methods. Extract samples did not show any significant cytotoxicity. Conclusions Extract samples of eucommiae cortex were obtained through freeze-drying and spray-drying. Neither change in index compound content nor difference in anti-inflammatory activity was observed between drying methods.

• The Korea Journal of Herbology
• /
• v.26 no.4
• /
• pp.39-47
• /
• 2011

Objective: The roots, leaves, flowers, stems and seeds of Cirsium japonicum var. ussuriense are often used in treatment of human diseases such as hemorrhage, blood congestion and inflammation. Focusing our attention on natural and bioavailable sources of antioxidants and anti-inflammation, we undertook to investigate the antioxidant and anti-inflammatory properties of Cirsium japonicum var. ussuriense used as a folk medicine in Korea. Methods: The extracts of the leaves, stems, flowers, seeds and roots from C. japonicum var. ussuriense were prepared by extracting with water or 80% ethanol. Total flavonoids and polyphenols were measured by a colorimetric assay. The free radical scavenging activity of the extract was analyzed by the DPPH (1,1-diphenyl-2-picryl hydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and Griess reagent assay. An oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The level of prostaglandin $E_2$ ($PGE_2$) was measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were measured by Western blot analysis. Results: Total flavonoid and polyphenol amounts of the leaves (CLE) and flowers (CFE) showed higher than those of the seed extract (CSE), stem extract (CSTE) and roots (CRE). CLE and CFE also showed the high antioxidant activities such as DPPH, NO-like and ABTS radical scavenging activity. An antioxidant activities of these water extracts showed higher than those of 80% ethanol extracts. We investigated the anti-inflammatory effects of CLE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. CLE significantly suppressed the levels of the inflammatory mediators such as NO and prostaglandin $E_2$ ($PGE_2$) in dose dependant. Furthermore, the levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with CLE extract in a dose dependent manner. Conclusions: These results suggest that CLE water extract has a higher anoxidant and anti-inflammatory activity, these properties may contribute to the oxidative and inflammatory related disease care.

• Journal of Convergence for Information Technology
• /
• v.9 no.12
• /
• pp.294-301
• /
• 2019

This study evaluated the antioxidant activity and the anti-inflammatory activity of ethanol extracted Pyracantha angustifolia (PE) or hot water extracted P. angustifolia (PW) using natural plant sources. In the DPPH and ABTS assay, the PE extracts showed the highest activity with an IC₅₀ of 3.78 ㎍/mL, IC₅₀ of 510.57 ㎍/mL, respectively. The total polyphenol content of PE extracts was 37.11±0.01 mgGAE/mL and PW extracts was 11.46±0.01 mgGAE/mL in a 1 mg/mL. The MTT assay showed no cytotoxicity at all concentration of two extracts in mouse macrophage cell line, RAW264.7. In addition, PE extracts strongly inhibited the production of NO, TNF-α cytokine secretion, and iNOS/TNF-α mRNA expression stimulated by LPS in RAW264.7 cells. Taken together, these results suggest that P. angustifolia, especially the ethanol extracts (PE), can be used as a cosmetic material containing natural antioxidant and anti-inflammatory properties.

• Nutrition Research and Practice
• /
• v.16 no.6
• /
• pp.685-699
• /
• 2022

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG) has long been known as a medicinal herb effective in various diseases, including bronchitis and asthma, but is still more widely used for food. Fermentation methods are being applied to increase the pharmacological composition of PG extracts and commercialize them with high added value. This study examines the hydrolyzed and fermented PG extract (HFPGE) fermented with Lactobacillus casei in RAW 264.7 cells, and investigates the effect of amplifying the immune and the probable molecular mechanism. MATERIALS/METHODS: HFPGE's total phenolic, flavonoid, saponin, and platycodin D contents were analyzed by colorimetric analysis or high-performance liquid chromatography. Cell viability was measured by the MTT assay. Phagocytic activity was analyzed by a phagocytosis assay kit, nitric oxide (NO) production by a Griess reagent system, and cytokines by enzyme-linked immunosorbent assay kits. The mRNA expressions of inducible nitric oxide synthase (iNOS) and cytokines were analyzed by reverse transcription-polymerase chain reaction, whereas MAPK and nuclear factor (NF)-κB activation were analyzed by Western blots. RESULTS: Compared to PGE, HFPGE was determined to contain 13.76 times and 6.69 times higher contents of crude saponin and platycodin D, respectively. HFPGE promoted cell proliferation and phagocytosis in RAW 264.7 cells and regulated the NO production and iNOS expression. Treatment with HFPGE also resulted in increased production of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, C-X-C motif chemokine ligand10, granulocyte-colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and monocyte chemoattractant protein-1, and the mRNA expressions of these cytokines. HFPGE also resulted in significantly increasing the phosphorylation of NF-κB p65, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. CONCLUSIONS: Taken together, our results imply that fermentation and hydrolysis result in the extraction of more active ingredients of PG. Furthermore, we determined that HFPGE exerts immunostimulatory activity via the MAPK and NF-κB signaling pathways.