• 동의생리병리학회지
• /
• 제35권1호
• /
• pp.15-21
• /
• 2021

Donggwaja (Benincasae Semen), the seed of Benincasa hispida (Thunb.) Cogn., has been used in Korean traditional medicine to control the body heat and water retention caused by various diseases. Both the symptoms targeted by the herbal medicine in clinic and studies with disease mouse models support the potential anti-inflammatory effect of Donggwaja. However, it is less understood how Donggwaja exerts its possible anti-inflammatory effect. Here, we present evidence that Donggwaja suppresses macrophage inflammatory reactions via expressing tumor necrosis factor a-induced protein 3 (TNFAIP3 or A20) and suppressing NF-kB activity. The ethanol extract of Donggwaja (EED) showed no toxicity when added to RAW 264.7 cells less than 100mg/ml. When treating the cells for 16 h, EED significantly suppressed the nuclear localization of NF-kB, suggesting that EED suppresses NF-kB activity. Concordantly, a semi-quantitative RT-PCR analysis showed that EED decreased the expression of prototypic pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-a, IL(interleukin)-6, and IL-1b. EED induced in RAW 264.7 cells the expression of A20, a ubiquitin modulator that suppresses inflammatory signaling cascades initiated from TLR4 and TNF and IL-1 receptors, while not affecting the induction of Nrf2, an anti-inflammatory factor that could suppress the effect of NF-kB. These results suggest that EED exerts its suppressive effect on inflammation, at least in part, by expressing anti-inflammatory factor A20 and suppressing pro-inflammatory factor NF-kB activity.

• The Plant Pathology Journal
• /
• 제37권6호
• /
• pp.632-640
• /
• 2021

Cucumber mosaic virus (CMV) and Pepper mild mottle virus (PMMoV) causes severe economic loss in crop productivity of both agriculture and horticulture crops in Korea. The previous surveys showed that naturally available biopolymer material - chitosan (CS), which is from shrimp cells, reduced CMV accumulation on pepper. To improve the antiviral activity of CS, it was synthesized to form phosphate cross-linked chitosan (PCS) and compared with the original CS. Initially, the activity of CS and PCS (0.01%, 0.05%, and 0.1% concentration) compound against PMMoV infection and replication was tested using a half-leaf assay on Nicotiana glutinosa leaves. The total number of local lesions represented on a leaf of N. glutinosa were counted and analyzed with phosphate buffer treated leaves as a negative control. The leaves treated with a 0.1% concentration of CS or PCS compounds exhibited an inhibition effect by 40-75% compared with the control leaves. The same treatment significantly reduced about 40% CMV accumulation measured by double antibody sandwich enzyme-linked immunosorbent assay and increased the relative expression levels of the NPR1, PR-1, cysteine protease inhibitor gene, LOX, PAL, SRC2, CRF3 and ERF4 genes analyzed by quantitative reverse transcriptase-polymerase chain reaction, in chili pepper plants.

• 대한의생명과학회지
• /
• 제27권4호
• /
• pp.216-222
• /
• 2021

Colorectal cancer (CRC) is major cancer with high incidence and mortality worldwide. It is known that most CRCs arise from precursor adenomatous polyps (APs). Recently, microRNA (miRNA) has been proposed as a biomarker for various cancers including CRC. In this study, the expression patterns of miR-29a in the whole blood (WB) of CRC, AP, and control groups were analyzed by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) to evaluate the expression level of miR-29a in patients with colorectal neoplasm (CRN) including CRC and AP. As a result, the relative expression of miR-29a was significantly decreased in the patients with CRN compared to the control group (P<0.001). The results were in agreement with previous in vitro cell studies and studies that used tissue and feces samples, suggesting that miR-29a in WB may be useful in demonstrating the status of colorectal tissue. Additionally, we divided the control group into healthy control (HC) without any colorectal symptoms and non-tumor control (NTC) with colorectal symptoms but without any CRN. And then the relative expression of miR-29a was also significantly decreased in the NTC group compared to the HC group (P<0.001). Therefore, our study revealed that miR-29a can differentiate patients with CRN from HC group, but they are also involved in the early stage of inflammatory response and cannot be specific biomarkers for CRN.

• 대한본초학회지
• /
• 제32권3호
• /
• pp.55-61
• /
• 2017

Objectives : The aim of this research was to determine the diverse effects of Lithospermi Radix Water Extract (LR) on human keratinocyte HaCaT cells, and to examine whether those effects could be applied to the human skin. Methods : We examined effect of LR on the cell viability of using the MTS assay in human keratinocyte HaCaT cells. The antioxidation effect of LR was analyzed relative to the well-known antioxidant resveratrol, using an ABTS assay. Quantitative RT-PCR analysis revealed that, in HaCaT cells, LR influenced the mRNA expression of tight-junction genes associated with skin moisturization. Furthermore, a wound-healing assay demonstrated altered cell migration in LR-treated HaCaT cells. Result : The cytotoxicity was confirmed to be higher in LR at a concentration of $800{\mu}g/m{\ell}$ using the MTS assay in HaCaT cells. In comparison to $100{\mu}M$ resveratrol, $1,600{\mu}g/m{\ell}$ LR showed either a similar or superior antioxidation effect. LR treatment in HaCaT cells reduced the mRNA expression levels of claudin 3, claudin 4, claudin 6, claudin 8, and ZO-2 to less than 0.80-fold, whereas JAM-A and Tricellulin mRNA expression level increased more than 1.33-fold. In addition, HaCaT cells migration was decreased to 83.9% by LR treatment. Conclusions : LR of antioxidation activity will have an anti-aging effect on the skin by reducing oxidative stress. Further studies are required to address the implications for human skin, given LR's effects of altering mRNA expression of tight junction-related gene and decreasing cell migration of HaCaT cells.

• 대한본초학회지
• /
• 제33권1호
• /
• pp.1-7
• /
• 2018

Objectives : Puerariae Radix extracts are a rich source of isoflavones that affect the activity of hepatic antioxidant enzymes. However, the effect of these extracts on hair growth in human dermal papilla (DP) cells is unknown. The purpose of this study is to compare the effects between juice and water extracts of Puerariae Radix on the mRNA expression levels of hair growth-related genes in dermal papilla (DP) cells. Methods : The antioxidant activity of juice and water extracts of Puerariae Radix was analyzed using an ABTS assay. The cytotoxicity was analyzed using the MTS assay in DP cells. mRNA expression levels of hair growth-related genes in dermal papilla (DP) cells were analyzed using quantitative RT-PCR analysis. Results : Juice and water extracts of Puerariae Radix showed strong antioxidant activity. The cytotoxicity was confirmed to be higher in the juice extract than the water extract, using the MTS assay on DP cells. The mRNA expression levels of CTNNB1, FGF7, and BMP6 were significantly increased after treatment with water extract, whereas the juice extract did not affect the expression of hair growth-related genes. Conclusions : Our study provides evidence that water extract of Puerariae Radix is effective at inducing hair growth, by promoting the mRNA expression levels of hair growth-related genes. Also in the future, studies should be conducted to investigate the effects of Puerariae radix extracts on the various hair growth mechanisms of dermal papilla cells.

• 대한본초학회지
• /
• 제34권2호
• /
• pp.25-32
• /
• 2019

Objectives : Gardeniae Fructus extract is used as a component of various cosmetics. However, the effect of the extract on the motility of keratinocytes has not been studied. The aim of this study is to investigate the inhibitory effect of ethanol extract of Gardeniae Fructus (GFET) or ethyl acetate extract of Gardeniae Fructus (GFEA) on oxidation and motility of human keratinocyte HaCaT cells. Methods : Antioxidant activity of Gardeniae Fructus extracts were determined by the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. To investigate the cytotoxicity of Gardeniae Fructus extracts, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed. The mRNA expression levels of tight junction related genes were analyzed using quantitative RT-PCR analysis. Cell migration assay was employed to determine the activity of Gardeniae Fructus extracts on motility of human keratinocyte HaCaT cells. Results : GFET and GFEA showed strong antioxidant activity. GFEA showed stronger cytotoxicity in HaCaT cells than GFET until $2.0mg/m{\ell}$ concentration. Cell migration assay demonstrated that GFET and GFEA decreased the motility of HaCaT cells. In addition, the mRNA expression level of claudin 8 among tight junction genes was significantly reduced by GFET or GFEA treatment. Conclusions : We investigated the physiological activities of the extracts of Gardeniae Fructus extracts on human keratinocytes by two different extraction methods. In addition, the mRNA expression level of claudin 8 among tight junction genes was significantly reduced by either GFET or GFEA treatment. This study provides basic information on the application of Gardeniae Fructus extract to cosmetics component.

• 대한본초학회지
• /
• 제34권1호
• /
• pp.99-107
• /
• 2019

Objectives : Cudrania tricuspidata (C. tricuspidata) is well-known traditional herbal remedy and its root, leaf and fruit were used for treatment of inflammation, tumor and painkilling. However, effect of C. tricuspidata fruit on tight junction is still unknown. The aim of this research was to determine the effect of C. tricuspidata fruit extract on human keratinocyte HaCaT cells. Methods : The antioxidant effects of water extract of C. tricuspidata (WECT) and ethanol extract of C. tricuspidata (EECT) were analyzed by using an ABTS assay. To confirm the cytotoxicity of WECT and EECT, MTS assay was performed. The mRNA expression levels of tight junction related genes were analyzed using quantitative RT-PCR analysis. Furthermore, dispase assay was used to investigate the alteration of cell-cell adhesion strength of EECT treated HaCaT cells. Results : WECT and EECT showed strong antioxidant activity. No obvious cytotoxicity was observed in both WECT and EECT until $2.0mg/m{\ell}$ concentration. The mRNA expression level of Claudin 6 were significantly increased by EECT treatment, whereas the WECT did not affect the expression of Claudin 6. Furthermore, EECT treatment enhances cell-cell adhesion strength. Conclusions : In this study, we investigated the physiological activities of the extracts of Cudrania tricuspidata fruit extracts on human keratinocytes by two different extraction methods. EECT might have an anti-aging activity on the skin by reducing oxidative stress. Moreover, it may be a useful ingredient in atopic dermatitis and skin-moisturizing, given its effects of altering Claudin 6 gene expression and enhancing cell-cell adhesion strength.

• Journal of Veterinary Science
• /
• 제21권3호
• /
• pp.50.1-50.16
• /
• 2020

Background: Porcine parvovirus (PPV) is a single-stranded DNA virus that causes porcine reproductive failure. It is of critical importance to study PPV pathogenesis for the prevention and control of the disease. NS1, a PPV non-structural protein, is participated in viral DNA replication, transcriptional regulation, and cytotoxicity. Our previous research showed that PPV can activate nuclear factor kappa B (NF-κB) signaling pathway and then up-regulate the expression of interleukin (IL)-6. Objectives: Herein, the purpose of this study is to determine whether the non-structural protein NS1 of PPV also has the same function. Methods: Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay, western blot, immunofluorescence assay and small interfering RNA (siRNA) were used. Results: Our findings demonstrated that PPV NS1 protein can up-regulate the expression levels of IL-6 and tumor necrosis factor-alpha in a dose-dependent manner. Moreover, PPV NS1 protein was found to induce the phosphorylation of IκBα, then leading to the phosphorylation and nuclear translocation of NF-κB. In addition, the NS1 protein activated the upstream pathways of NF-κB. Meanwhile, TLR2-siRNA assay showed TLR2 plays an important role in the activation of NF-κB signaling pathway induced by PPV-NS1. Conclusions: These findings indicated that PPV NS1 protein induced the up-regulated of IL-6 expression through activating the TLR2 and NF-κB signaling pathways. In conclusion, these findings provide a new avenue to study the innate immune mechanism of PPV infection.

• Journal of Ginseng Research
• /
• 제47권3호
• /
• pp.429-439
• /
• 2023

Background: The incidence and clinical importance of nonalcoholic fatty liver disease (NAFLD) has emerged. However, effective therapeutic strategies for NAFLD have yet to be found. Panax ginseng (P. ginseng) is a traditional herb in Eastern Asia with therapeutic effects in many chronic disorders. However, the precise effects of ginseng extract on NAFLD are currently unknown. In present study, the therapeutic effects of Rg3-enriched red ginseng extract (Rg3-RGE) on the progression of NAFLD were explored. Methods: Twelve-week-old C57BL/6 male mice were fed a chow or western diet supplemented with high sugar water solution with or without Rg3-RGE. Histopathology, immunohistochemistry, immunofluorescence, serum biochemistry, western blot analysis, and quantitative RT-PCR were used for in vivo experiment. Conditionally immortalized human glomerular endothelial cell (CiGEnC) and primary liver sinusoidal endothelial cells (LSECs) were used for in vitro experiments. Results: Eight weeks of Rg3-RGE treatment significantly attenuated the inflammatory lesions of NAFLD. Furthermore, Rg3-RGE inhibited the inflammatory infiltrate in liver parenchyma and the expression of adhesive molecules to LSECs. Moreover, the Rg3-RGE exhibited similar patterns on the in vitro assays. Conclusion: The results demonstrate that Rg3-RGE treatment ameliorates NAFLD progression by inhibiting chemotaxis activities in LSECs.

• 동의생리병리학회지
• /
• 제37권1호
• /
• pp.14-18
• /
• 2023

Jukyeo (竹茹; Bambusae Caulis in Taeniam: BCT) is an herbal medicine made from the inner part of the bamboo stem of Phyllostachys nigra Munro var. henonis Stapf ex Rendle or Phyllostachys bambusoides Siebold et Zuccarini. Although medical literature published in China and Korea decades ago introduced BCT as a remedy for reducing vomiting, fever, and hematemesis, decoctions containing BCT as a main herb (君藥) in Korea have been approved only for treating neurologic symptoms. Here, we discuss the anti-inflammatory function of BCT. Combined with the clinical usage of a BCT-containing decoction in treating inflammatory diseases in Japan, we raise the possibility of repurposing the BCT-containing decoctions for treating inflammatory diseases. The anti-inflammatory activity of BCT was mainly assessed by using RAW 264.7 cells. The regulation of NF-κB, Nrf2 and A20 activities was determined by western blot and quantitative RT-PCR. The list of decoctions containing BCT currently approved in Korea was obtained from the Korean Ministry of Food and Drug Safety (KFDA). BCT suppressed the activity of pro-inflammatory factor NF-κB elicited by LPS, activated an anti-inflammatory factor Nrf2, and induced A20 that is known to block several inflammatory pathways simultaneously, suggesting that BCT can suppress inflammation via at least 3 different pathways. KFDA approved 11 decoctions containing BCT as a major herb, including Gamiondam-tang (加味溫膽湯; GOT), for treating neurologic disorders. Interestingly, Jukyeoondam-tang (竹茹溫膽湯; JOT), whose composition is almost identical to GOT except for one herb, has been used to treat inflammatory pulmonary disorders including Covid-19 pulmonary infection in Japan Given the anti-inflammatory function of BCT evidenced by medical literature and experimental results and the clinical usage of JOT in treating inflammatory pulmonary disorders, we suggest a repurposing and extension of the BCT-containing decoctions approved in Korea to treating inflammatory diseases.