• The Korea Journal of Herbology
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• v.33 no.2
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• pp.69-77
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• 2018

Objectives : Daesiho-tang (DSHT) has been widely used in the treatment of cerebral infarct in traditional medicine. However, there was not report on the anti-obesity-related diseases efficacy of DSHT. In this study, we investigated the effects for the new formulation of DSHT, on the adipocyte differentiation cycle in 3T3-L1 cells. Methods : 3T3-L1 cells were treated with DSHT (50, 100, $200{\mu}g/m{\ell}$) during differentiation for 6 days. Also, the inhibitory effect of DSHT against 3T3-L1 adipogenesis was evaluated in various stage of adipogenesis such as early (0-2day), intermediate (2-4day), and terminal stage (4-6day). The accumulation of lipid droplets was determined by Oil Red O staining. and, the expressions of genes related to adipogenesis were measured by RT-PCR and Western blot analyses. Results : DSHT showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affect cell toxicity as assessed by measuring fat accumulation and adipogenesis. In addition, DSHT significantly reduced the expression levels of several adipocyte marker genes including proliferator activated $receptor-{\gamma}$ ($PPAR-{\gamma}$) and CCAAT/ enhancer-binding $protein-{\alpha}$ ($C/EBP-{\alpha}$). Also, the anti-adipogenic effect of DSHT was strongly limited in the intermediate (2-4 day), terminal stage (4-6 day) of 3T3-L1 adipogenesis. In addition, the DSHT treatment down- regulated mRNA expression levels of $PPAR-{\gamma}$,, $C/EBP-{\alpha}$ in mature 3T3-L1 adipocytes. Conclusions : These results suggest that, the ability of DSHT has inhibited overall adipogenesis and lipid accumulation in the 3T3-L1 cells. The new formulation of DSHT may be a promising medicine for the treatment of obesity and related metabolic disorders.

• The Korea Journal of Herbology
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• v.27 no.6
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• pp.37-42
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• 2012

Objectives : Angelica Gigantis Radix is prescribed as the root of different Angelica species on the pharmacopoeia in Korea, Japan and China. Chemical components and their biological activities were also different according to their species. A study for the development of simple method to compare Angelica roots was needed. In order to classify them, the methods such as DNA sequencing analysis and taste sensor were applied to three Angelica species like Angelica gigas, Angelica acutiloba and Angelica sinensis. Methods : PCR amplification of intergenic transcribed spacer (ITS) region was performed using ITS1 and ITS4 primer from nine Angelica roots, and then nucleotide sequence was determined. Taste pattern of samples were measured using the taste-sensing system SA402B equipped with a sensing unit, which consists of artificial lipid membrane sensor probes of anionic bitterness, astringency, saltiness, umami, and cationic bitterness (C00, AE1, CT0, AAE, and AN0, respectively). Results : As a result of comparing the similarity of the ITS region sequences, A. sinensis was discriminated from the others (A. gigas and A. acutiloba). Equally this genetic result, A. gigas and A. acutiloba showed similar taste pattern as compared to A. sinensis. Sourness, bitterness, aftertaste of bitterness, astringency, and aftertaste of astringency of A. sinensis were significantly high as compared with A. gigas and A. acutiloba. In contrast, richness was significantly low. Conclusions : These taste pattern can be used as a way of comparison of Angelica species and this technic could be applied to establish a taste pattern marker for standardization of herbs in various purposes.

• The Korea Journal of Herbology
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• v.32 no.2
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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 $TNF-{\alpha}$.

• The Korea Journal of Herbology
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• v.32 no.2
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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.

• The Korea Journal of Herbology
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• v.33 no.3
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• pp.25-35
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• 2018

Objectives : Lepidii seu Descurainiae Semen has been frequently adulterated with the seeds of several inauthentic plant species. However, the accurate identification of these plant seeds is very difficult. To develop a reliable genetic authentication tool for Lepidii seu Descurainiae Semen, we analyzed matK sequence. Methods : To obtain the matK sequences of plant materials, genomic DNA was extracted from 24 samples and PCR amplification was carried out using matK-AF/matK-8R universal primer set and matK-LDSF/matK-LDSR primer set. For identifying species-specific nucleotides and phylogenetic analysis, matK regions were sequenced and comparatively analyzed by the ClustalW and Maximum Likelihood method. Results : We developed a new primer set to amplify matK region in Lepidii seu Descurainiae Semen and closely related plant samples. From the comparative analysis of matK sequences, we identified species-specific marker nucleotides for D. sophia, L. apetalum, L. latifolium, E. cheiranthoides, E. macilentum, and D. nemorosa, respectively. Furthermore, phylogenetic analysis revealed clear classification depending on the species. These results indicated that the matK sequence obtained a new primer set in this study was useful to identify Lepidii seu Descurainiae Semen in species level. Conclusions : We developed a primer set and identified species-specific marker nucleotides enough to distinguish authentic Lepidii seu Descurainiae Semen and adulterants at the species level based on the matK sequences. These genetic tool will be useful to prevent adulteration and to standardize the quality of Lepidii seu Descurainiae Semen.

• The Korea Journal of Herbology
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• v.33 no.2
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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.

• The Korea Journal of Herbology
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• v.34 no.1
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• pp.23-31
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• 2019

Objectives : The aim of this study was to investigate the effect for allergic-inflammation of Fritillariae Thunbergii Bulbus (FTB) on HaCaT cells and RBL2H3 cells. Methods : To investigate the effects of FTB for anti-inflammation in HaCaT cells, the cells were pretreated with FTB for 1h and then stimulated with $TNF-{\alpha}/IFN-{\beta}$ for 24h. Then thymus and activation-regulated chemokine (TARC) and Macrophage-derived chemokine (MDC) levels were analyzed with ELISA kit. Also to investigate the effect of skin barrier protein, the cells were treated with FTB of various concentrations, and then cells were harvested, expressions of skin barrier protein were measured with RT-PCR. To investigate the effects of FTB for anti-allergy in RBL2H3 cells, the cells were pre-treated with FTB for 1h, and then stimulated with A23187 for 30 min. ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ were measured using cultured media. The cells were harvested to analyze the mechanism of the effect for FTB via Western blot. Results : FTB did not show cytotoxicity in HaCaT and RBL2H3. In HaCaT cells, FTB significantly suppressed the expression of TARC, MDC at a dose-dependent manner and markedly increased formation of the skin barrier proteins. In RBL2H3 cells, FTB decreased release of the ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ in RBL2H3 through inhibition of the phosphorylation of JNK and p38, which are include in the signaling mechanism of MAPK Conclusion : These results indicate that FTB has an anti-inflammatory effect on the allergic response through blocking MAPK pathway. This suggest that FTB could be a therapeutic agent for allergic response.

• The Korea Journal of Herbology
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• v.34 no.1
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• pp.75-80
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• 2019

Objectives : Dictamni Radicis Cortex (DRC) has been used as an important traditional medicine for inflammation and fungal diseases. However, the protective effect of DRC water extract on acute pancreatitis (AP) has not been deeply reported. Therefore, we aimed to evaluate the protective effects of DRC water extract on cerulein-induced AP. Methods : AP was induced via intraperitoneal injection of supramaximal concentrations of stable cholecystokinin analogue cerulein ($50{\mu}g/kg$) every hour for 6 times. DRC water extract (0.05, 0.1, or 0.2 g/kg) or saline was administrated intraperitoneally 1 h before to the first injection of cerulein. The mice were sacrificed at 6 h after the final cerulein injection. Pancreas was rapidly removed for histochemical examination and myeloperoxidase (MPO) assay. In addition, polymerase chain reaction (PCR) was performed to examine mRNA levels of proinflammatory cytokines such as Interleukin $(IL)-1{\beta}$, IL-6 and Tumor necrosis factor $(TNF)-{\alpha}$. Results : Administration of DRC water extract significantly inhibited the pancreatic weight to body weight ratio, pancreas histological damages and increase of pancreatic MPO activity during cerulein-induced AP. In addition, increased pancreatic mRNA levels of $IL-1{\beta}$, IL-6 but not $TNF-{\alpha}$ were significantly inhibited by treatment of DRC water extract against cerulein-induced AP. Conclusions : In conclusion, we have revealed that pre-treatment of DRC water extract reduces the severity of cerulein-induced AP. Accordingly, our results could give a clinical basis that DRC could be used as a drug or agent to prevent AP.

• Korean Journal of Acupuncture
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• v.39 no.2
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• pp.43-53
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• 2022

Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.

• Journal of Marine Life Science
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• v.6 no.2
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• pp.73-79
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• 2021

Bisphenol A is a representative endocrine disruptor and continuously detected in aquatic environment due to wide use, resulting in adverse effects on growth, development, and reproduction in diverse organisms as well as human. Structural analogs have been developed to substitute BPA are also suspected to have endocrine disrupting effects. In the present study, the time-dependent expression patterns of ecdysteroid synthesis (nvd, cyp314a1), receptors (EcRA, EcRB, USP, ERR), and downstream signaling pathway - related genes (HR3, E75, Vtg, VtgR) were investigated using quantitative real time reverse transcription polymerase chain reaction (qRT-PCR) in the brackish water flea Diaphanosoma celebensis exposed to Bisphenol analogs (BPs; BPA, BPF, and BPS) for 6, 12, and 24 h. As results, the expression of nvd, cyp314a1, EcRs, USP, ERR and E75 mRNA was upregulated at 6 h exposure to BPF, which is earlier than BPA and BPS (12 h). On the other hand, HR3, E75 and VtgR mRNA levels were elevated at 6 h earlier at BPS and BPF than at BPA (12 h), but Vtg mRNA level was slightly changed within 24 h. These findings suggest that like BPA, BPF and BPS can also modulate the transcription of ecdysteroid pathway - related genes with different mechanisms, and have a potential as endocrine disruptors. This study will provide a better understanding the molecular mode of action of bisphenols on ecdysteroid pathway in the brackish water flea.