• 대한본초학회지
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• 제28권5호
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• pp.7-11
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• 2013

Objectives : Seungmagalgeun-tang (SMGGT) is traditional medicine widely used for inflammatory disease and flu. But SMGGT exhibits potent anti-inflammatory activity with an unknown mechanism. To elucidate the molecular mechanisms of SMGGT water extract on pharmacological and biochemical actions in inflammation, we examined the effect of SMGGT on pro-inflammatory mediators in Phorbol-12-myristate-13-acetate (PMA)+A23187-stimulated mast cells. Methods : In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to measure the activation of MAPKs. Cells were treated with SMGGT 1 h prior to the addition of 50 nM of PMA and $1{\mu}M$ of A23187. Cell viability was measured by MTS assay. The investigation focused on whether SMGGT inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8) and mitogen-activated protein kinases (MAPKs) in PMA+A23187-stimulated mast cells. Results : SMGGT has no cytotoxicity at examined concentration (100, 250, and $500{\mu}g/ml$). Also, gene expression of IL-6 and IL-8 in HMC-1 cells stimulated by PMA+A23187 was down regulated by SMGGT. Furthermore, SMGGT suppressed the PMA+A23187-induced phosphorylation of extracellular signal-regulated kinase (ERK) and c-jun N-terminal Kinase(JNK). But, SMGGT could not regulate phosphorylation of p38 MAPK. Conclusions : These results suggest that SMGGT has inhibitory effects on PMA+A23187-induced IL-6 and IL-8 production. These inhibitory effects occur through blockades on the phosphorylation of ERK and JNK.

• 대한본초학회지
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• 제32권3호
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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.

• 대한본초학회지
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• 제32권3호
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• pp.45-53
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• 2017

Objective : Nelumbo nucifera, its rhizome and semen have been used as a traditional medicine which was studied on antioxidant, hepatoprotective effect, anti-obesity and the others. However, Nelumbinis Flos have not studied. We investigated protective effects on oxidative DNA damage and anti-inflammatory effects of Nelumbinis Flos. Methods : The antioxidant activity was conducted by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay and reducing power assay. Total phenolic content was analyzed. Also, phenolic compounds were detected by HPLC/UV. The inhibitory effect on oxidative DNA damage was determined using ${\Phi}X-174$ RF I plasmid DNA cleavage assay. The anti-inflammatory effect of Nelumbinis Flos was measured by the amount of nitric-oxide (NO) produced and protein levels of iNOS, and COX-2 in LPS induced RAW 264.7 cells. Results : The results of DPPH and ABTS radical scavenging activity at $200{\mu}g/m{\ell}$ of extraction were $97.02{\pm}0.88%$ and $96.42{\pm}0.25%$. Reducing power (fold of L-ascorbic acid as control) was $100.14{\pm}0.31$ at $200{\mu}g/m{\ell}$. Total phenol content was $8.70{\pm}0.02mg/g$. Chlorogenic acid, catechin and epicatechin were found by HPLC. Nelumbinis Flos has inhibitory effect in dose-manner against oxidative DNA damage. In addition, it showed the anti-inflammatory effect by suppression of NO production as well as protein levels of iNOS, and COX-2. Conclusion : This study suggested that Nelumbinis Flos showed potential antioxidant and suppression activities of various factors were related in NO produced. Therefore, Nelumbinis Flos as natural plant resources that may help reduce inflammation and alleviate DNA damage.

• 대한본초학회지
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• 제34권1호
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• pp.51-57
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• 2019

Objectives : Nypa fruticans Wurmb. (NF) have been used as a traditional medicine to treat inflammatory diseases in East-South Asia. However, it is largely undiscovered whether NF water extract could exhibit anti-inflammatory activities against tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced inflammatory responses on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the anti-inflammatory activity of NF water extract on TNF-${\alpha}$-induced inflammatory responses in HaCaT cells. Methods : To investigate the anti-inflammatory activites of NF water extract in HaCaT cells, the inflammatory model of HaCaT cells was established under a suitable concentration (10 ng/ml) of human TNF-${\alpha}$ (hTNF-${\alpha}$). HaCaT keratinocyte cells were pre-treated with NF water extract for 1 h, and then stimulated with hTNF-${\alpha}$. Then, the cells were harvested to measure the inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$), and pro-inflammatory cytokine including TNF-${\alpha}$ and interleukin (IL)-6. In addition, we examined the inhibitory mechanisms of NF, mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha ($I{\kappa}-B{\alpha}$) Results : The treatment of NF inhibited the hTNF-${\alpha}$-induced elevation of iNOS, COX-2, and $PGE_2$ in HaCaT cells. In addition, NF treatment inhibited the hTNF-${\alpha}$-induced elevation of TNF-${\alpha}$ and IL-6. Furthermore, NF treatment inhibited the activation of MAPKs but not degradation of $I{\kappa}-B{\alpha}$. Conclusions : Taken together, our result suggest that treatment of NF could inhibit the hTNF-${\alpha}$-induced inflammatory responses via deactivation of MAPKs in HaCaT cells. This study could suggest that NF could be a beneficial agent to prevent skin damage or inflammation.

• 대한본초학회지
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• 제33권6호
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• pp.71-78
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• 2018

Objectives : This study was fulfilled to investigate nominee materials as anti-inflammatory agent from ethanol extract of Chinese medicinal plants in Yanjin. Among the 20 candidates, we selected most effective one, the ethanol extract of Cicuta virosa L. (CVL). The mechanism underlying the anti-inflammatory effects of CVL is not clearly identified as yet. Accordingly, we clarified the anti-inflammatory effects of CVL and its underlying molecular mechanisms in LPS-stimulated RAW 264.7 macrophages. Methods : RAW264.7 macrophages were incubated with CVL (12.5, 25, or $50{\mu}M$) and/or lipopolysaccharide (LPS) ($1{\mu}g/m{\ell}$). Cytotoxicity was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and the level of nitric oxide (NO) production was measured with Griess reagent. The prostaglandin $E_2$ ($PGE_2$) production was measured with enzyme immunoassay kits and the protein expression of inducible nitric oxide synthase (iNOS) was determined using Western blot analysis. Results : Among the 20 ethanol extract of Chinese medicinal plants of Yanjin tested, CVL significantly reduced the production of NO in a dose-dependent manner via inhibition the protein expressions of iNOS without cytotoxicity on the LPS-stimulated RAW 264.7 macrophages. In addition, CVL also effectively declined the production of $PGE_2$ in LPS-simulated RAW 264.7 macrophages. Conclusions : Taken together, these data presented in this study demonstrate that CVL possesses anti-inflammatory activity by suppressing the production of pro-inflammatory mediators NO and $PGE_2$, and pro-inflammatory protein iNOS expression in LPS-stimulated RAW 264.7 macrophages.

• 대한본초학회지
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• 제28권4호
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• pp.71-76
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• 2013

Objectives : The biological activities and compound contents of herbal medicine vary depending on manufacturing processes. In this study, we compared anti-inflammatory effects and compound contents of three kinds of multi-herbal extract HT008 produced by different manufacturing processes in order to determine chemical and biological equivalence. Methods : HT008 was produced by three different manufacturing methods: 1. Freeze dried extract of Eleutherococcus senticosus, Scutellaria baicalensis and Angelica sinensis (HT008 FD), 2. Spray dried extract of E. senticosus and S. baicalensis combined with reflux extract of A. sinensis (HT008 SD), 3. Spray dried extract of E. senticosus and S. baicalensis combined with supercritial fluid extract of A. sinensis (HT008 SF). Anti-inflammatory effects were evaluated using acetic acid induced pain model and ${\lambda}$-carageenan induced paw edema model. Compound contents were evaluated by HPLC quantitative analysis of standard compounds of HT008, eleutheroside E, baicalin, z-ligustilide. Results : HT008 FD, HT008 SD and HT008 SF significantly decreased acetic acid induced pain index and ${\lambda}$-carrageenan induced paw edema volume compared with that of control group. There was no significant difference in efficacy among the HT008 FD, HT008 SD and HT008 SF. Standard compound contents of HT008 FD, HT008 SD and HT008 SF were quantified within the range of Korean pharmacopoeia or other research. Conclusions : Three different manufacturing methods of multi-herbal extracts have been developed without noticeable difference in the efficacy or compound contents. The results might be used to establish manufacturing process and industrialization of herbal extracts.

• 대한본초학회지
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• 제32권2호
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• pp.77-85
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• 2017

Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.

• 대한본초학회지
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• 제32권2호
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• pp.107-114
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• 2017

Objectives : Tribulus terrestris $Linn{\acute{e}}$ (Tribuli Fructus; TF) has been used to treat hypochondrium, agalactia, nebula, itching and vitiligo in traditional Korean medicine. In this study, we investigated the effects of TF 30% ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : TF extract was prepared by 30% ethanol. RBL-2H3 cells, a rat mast cell line, were treated with TF extract at different concentrations for 1 hr and then stimulated with DNP-IgE/HSA for indicated times. Cell viability was measured by WST-1 assay. The expression of inflammatory cytokines (IL-4, IL-13 and $IFN-{\gamma}$) mRNA was determined by reverse transcriptase-PCR, and the phosphorylation of ERK1/2, p38 and JNK MAP kinases (MAPKs) was determined by Western blot. The nuclear expression of $NF-{\kappa}B$ p65 in the cells was detected by Western blot and immunocytochemistry, respectively. Results : The treatment of TF extract at 0.1 and $0.2mg/m{\ell}$ significantly decreased the expression of IL-4 and IL-13 mRNA in IgE-stimulated RBL-2H3 mast cells, while significantly increased the expression of $IFN-{\gamma}$ mRNA. TF extract treatment was also inhibited the phosphorylation of ERK1/2, p38 and JNK MAPKs in IgE-stimulated RBL-2H3 mast cells in a dose-dependent manner. In addition, TF extract significantly blocked the translocation of $NF-{\kappa}B$ p65 into the nuclear of cells after IgE stimulation. Conclusions : These results indicate that TF extract inhibits inflammatory response in IgE-stimulated mast cells through blocking MAPKs/$NF-{\kappa}B$ pathway. This suggests that TF extract has an anti-inflammatory activity in mast cell activation.

• 대한본초학회지
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• 제33권2호
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• pp.19-28
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• 2018

Objectives : Epimedii Herba has been frequently used in Korean Traditional Medicine to treat impotence, spermatorrhoea, exophthalmos, and forgetfulness. Present study investigated anti-inflammatory effects of Epimedii Herba water extract (EWE) and attempted to elucidate molecular mechanisms involved. Methods : To explore anti-inflammatory effects of EWE, RAW 264.7 cells, a murine macrophage cell line, were pretreated with $10-100{\mu}g/m{\ell}$ of EWE, and then subsequently exposed to $1{\mu}g/m{\ell}$ of lipopolysaccharide (LPS). Levels of nitric oxide (NO), interleukin-6, $interleukin-1{\beta}$, and tumor necrosis $factor-{\alpha}$ were monitored in the medium. Expression levels of inducible nitric oxide synthase and cyclooxygenase-2 were determined by immunoblot and real-time PCR analyses. Signaling pathways related with nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinases were monitored to elucidate molecular mechanisms involved. Finally, the role of three flavonoid compounds in EWE on LPS-mediated NO production were investigated. Results : In conditioned medium, pretreatment of EWE ($100{\mu}g/m{\ell}$) significantly inhibited LPS-stimulated NO and pro-inflammatory cytokine production. In addition, EWE attenuated the expressions of inducible nitric oxide synthase and cyclooxygenase-2 by LPS. EWE prevented the phosphorylation and degradation of inhibitory ${\kappa}B{\alpha}$, nuclear translocation of $NF-{\kappa}B$, and DNA binding of $NF-{\kappa}B$, while EWE did not change the phosphorylation of mitogen-activated protein kinases by LPS. Moreover, icariin, icaritin, and quercetin partly, but significantly, inhibited the LPS-stimulated NO production. Conclusions : These results suggest that EWE has an ability to prevent inflammation in macrophages through inhibition of $NF-{\kappa}B$ signaling pathway.

• 대한본초학회지
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• 제33권6호
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• pp.35-42
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• 2018

Objectives : The aim of this study was to investigate whether the extract of Coptidis Rhizoma inhibits inflammation in chronic cold stress (CCS)-exposed mice or not. Methods : Coptidis Rhizoma extract (CRE) was made by reflux with distilled water. Male ICR mice (7 weeks old) were divided randomly into 5 groups: (1) control, (2) CCS, (3) CCS+CRE 100 mg/kg, (4) CCS+CRE 300 mg/kg, (5) CCS+CRE 1,000 mg/kg groups. Mice were orally administered once a day for 14 days starting from 1 day before CCS. Group (2)-(5) were exposed to CCS conditions that maintained at $4^{\circ}C$ for 2 h once a day for 14 days. The levels of serum cortisol and hypothalamic prostaglandin E1 (PGE1) and PGE2 were measured by enzyme-linked immunosorbent assay kit. The expression levels of several pro-inflammatory factors like heat shock protein 70 (HSP70), c-fos, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) were measured by western blot analysis in mouse hypothalamus. Results : Oral administration of CRE 1,000 mg/kg significantly suppressed the increase of serum cortisol levels in mice exposed to CCS. CCS-exposed mice had significantly increased the expression of HSP70, c-fos, and NF-kB in hypothalamus, while CRE treatment significantly attenuated the elevation of these pro-inflammatory factors. The ratio of PGE2/PGE1 was also higher in CCS-exposed mice than control group. CRE treatment significantly reduced the increase of PGE2/PGE1 ratio induced by CCS. Conclusion : These findings suggest that Coptidis Rhizoma may work as a potential agent to modulate inflammatory responses under the condition of cold adaptation formed by CCS.