• The Korean Journal of Physiology and Pharmacology
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• v.26 no.5
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• pp.335-345
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• 2022

Pyroptosis is an inflammatory form of programmed cell death that is linked with invading intracellular pathogens. Cardiac pyroptosis has a significant role in coronary microembolization (CME), thus causing myocardial injury. Tanshinone IIA (Tan IIA) has powerful cardioprotective effects. Hence, this study aimed to identify the effect of Tan IIA on CME and its underlying mechanism. Forty Sprague-Dawley (SD) rats were randomly grouped into sham, CME, CME + low-dose Tan IIA, and CME + high-dose Tan IIA groups. Except for the sham group, polyethylene microspheres (42 ㎛) were injected to establish the CME model. The Tan-L and Tan-H groups received intraperitoneal Tan IIA for 7 days before CME. After CME, cardiac function, myocardial histopathology, and serum myocardial injury markers were assessed. The expression of pyroptosis-associated molecules and TLR4/MyD88/NF-κB/NLRP3 cascade was evaluated by qRT-PCR, Western blotting, ELISA, and IHC. Relative to the sham group, CME group's cardiac functions were significantly reduced, with a high level of serum myocardial injury markers, and microinfarct area. Also, the levels of caspase-1 p20, GSDMD-N, IL-18, IL-1β, TLR4, MyD88, p-NF-κB p65, NLRP3, and ASC expression were increased. Relative to the CME group, the Tan-H and Tan-L groups had considerably improved cardiac functions, with a considerably low level of serum myocardial injury markers and microinfarct area. Tan IIA can reduce the levels of pyroptosis-associated mRNA and protein, which may be caused by inhibiting TLR4/MyD88/NF-κB/NLRP3 cascade. In conclusion, Tanshinone IIA can suppress cardiomyocyte pyroptosis probably through modulating the TLR4/MyD88/NF-κB/NLRP3 cascade, lowering cardiac dysfunction, and myocardial damage.

• Nutrition Research and Practice
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• v.16 no.5
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• pp.549-564
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• 2022

BACKGROUND/OBJECTIVES: Oxidative stress is caused by an imbalance between harmful free radicals and antioxidants. Long-term oxidative stress can lead to an "exhausted" status of antioxidant defense system triggering development of metabolic syndrome and chronic inflammation. Green perilla (Perilla frutescens) is commonly used in Asian cuisines and traditional medicine in southeast Asia. Green perilla possesses numerous beneficial effects including anti-inflammatory and antioxidant functions. To investigate the potentials of green perilla leaf extract (PE) on oxidative stress, we induced oxidative stress by high-fat diet (HFD) in aging mice. MATERIALS/METHODS: C57BL/6J male mice were fed HFD continuously for 53 weeks. Then, mice were divided into three groups for 12 weeks: a normal diet fed reference group (NDcon), high-fat diet fed group (HDcon), and high-fat diet PE treated group (HDPE, 400 mg/kg of body weight). Biochemical analyses of serum and liver tissues were performed to assess metabolic and inflammatory damage and oxidative status. Hepatic gene expression of oxidative stress and inflammation related enzymes were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: PE improved hepatopathology. PE also improved the lipid profiles and antioxidant enzymes, including hepatic glutathione peroxidase (GPx) and superoxide dismutase (SOD) and catalase (CAT) in serum and liver. Hepatic gene expressions of antioxidant and anti-inflammatory related enzymes, such as SOD-1, CAT, interleukin 4 (IL-4) and nuclear factor erythroid 2-related factor (Nrf2) were significantly enhanced by PE. PE also reduced the levels of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) in the serum and liver; moreover, PE suppressed hepatic gene expression involved in pro-inflammatory response; Cyclooxygenase-2 (COX-2), nitric oxide synthase (NOS), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6). CONCLUSIONS: This research opens opportunities for further investigations of PE as a functional food and possible anti-aging agent due to its attenuative effects against oxidative stress, resulting from HFD and aging in the future.

• Journal of Physiology & Pathology in Korean Medicine
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• v.36 no.5
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• pp.181-186
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• 2022

Atractylodes macrocephala (AM) and Amomum villosum (AV) are the most common herbs in Korean Medicine to treat digestive diseases. In this study, we investigated the cholesterol lowering effects of mixtures of AM and AV extracts on high cholesterol diet (HCD) induced dyslipidemia mouse model. We classified animals into six different groups; Group 1: Normal diet, Group 2: HCD, Group 3: AV extracts : AM extracts (1:1) (200 mg/kg) + HCD, Group 4: AV extracts : AM extracts (1:2) (200 mg/kg) + HCD, Group 5: AV extracts : AM extracts (1:3) (200 mg/kg) + HCD, Group 6: Simvastatin 40 mg/kg + HCD. After 4 weeks of oral administration of respective drugs, we checked body, liver and epididymal fatweights along with liver and serum triacylglyceride (TG) concentration, total and low density lipoprotein (LDL) cholesterol in serum. Moreover, 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase (HMGCR), LDL receptor (LDLR), and sterol regulatory element-binding protein 2 (SREBP2) were detected by RT PCR or western blot analysis. The overall results showed that mixtures of AM and AV extracts inhibited HCD-induced increases of total cholesterol and LDL cholesterol in serum. Those effects seem to be caused by AM and AV extracts through inhibition of HMGCR expression. And thus blood cholesterol is induced into the liver by increasing LDLR expression, which is regulated by SREBP2 transcrption factor. The cholesterol lowering effects and mechanism of mixtures of AM and AV extracts was similar to the statin. We have identified the potential mixtures of AM and AV extracts as a new treatment for dyslipidemia.

• The Korea Journal of Herbology
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• v.38 no.5
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• pp.9-19
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• 2023

Objectives : Osteoporosis is a systemic skeletal disorder characterized by reduced bone mineral density and increased risk of fractures. Bisphosphonates and selective estrogen receptors, which are bone resorption inhibitors that are currently widely used as osteoporosis treatments, show serious side effects when administered for a long time. Research on bone resorption inhibitors that complement the problems of existing treatments is needed. The purpose of this study was to investigate the effect of inhibiting osteoclast differentiation and activity on the tuberous root of Ampelopsis japonica (Thunb.) Makino (AM). Methods : After extracting AM using distilled water and ethanol, the inhibitory effects of the two solvents on osteoclast differentiation were compared using the RANKL-induced in vitro experimental model and the TRAP assay kit. The impact of AM on bone resorption was investigated through the pit formation assay, and its effect on F-actin formation was assessed through fluorescent staining. Additionally, protein and mRNA expression levels of osteoclast differentiation markers (NFATc1, c-Fos, TRAP and ATP6v0d2) and resorption markers (MMP-9, CTK, and CA2) were analyzed via western blot and RT-PCR. Results : AM treatment significantly decreased the number of TRAP-positive cells and pit formation area. Furthermore, AM suppressed both the protein and mRNA expression of NFATc1 and c-Fos, key transcription factors involved in osteoclast differentiation and it downregulated the expression of osteoclast-associated genes such as TRAP, CTK, MMP-9, CA2, and ATP6v0d2. Conclusions : These results suggest that AM can inhibit bone resorption and osteoclast differentiation, indicating its potential for use in the treatment and prevention of osteoporosis.

• The Korea Journal of Herbology
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• v.24 no.3
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• pp.59-68
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• 2009

Objectives : Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods : PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results : In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterant Bupleurum species.

• The Korea Journal of Herbology
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• v.24 no.1
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• pp.179-189
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• 2009

Objectives : Coptidis Rhizoma (Coptis japonica MAKINO; CR) is a well known crude drug as antimicrobial, antibacterial, anti-inflammatory, antioxidant activity. However, there is no study of the effect of CR on brain infarction and it's mechanism. The aim of this study was to investigate the effects on ischemic stroke induced by photothrombotic infarction by evaluating the functional & neuronal recovery after brain infarction. Materials & Methods : Male Sprague-Dawley rats (250-300 g) were induced photothrombotic brain infarction on sensorimotor cortex, and brain infarction volume by image J software (NIH, USA) after Nissl stain, also single pellet reaching task as a functional motor recovery were observed. After orally pretreated by CR (500 mg/kg) or normal saline as a sham control before 7 days from the time of photothrombotic infarction, rats were sacrificed. After then we analysed anti-inflammatory cytokines (TNF-$\alpha$, IL-6, IL-1$\beta$), by RT-PCR and ELISA method, and immunohistochemistry (GFAP, connexin-43) as a marker of neural plasticity. Results : CR (100, 250, 500 mg/kg) decreased the infarction volume dose-dependently, however the effect of 500mg/kg of CR (CR 500) showed the best (P=0.051). Also, CR 500 decreased the infarction volume time-dependently, the most effective time was 3-7 days after stroke. Photothrombosis increased inflammatory cytokines after infarction, CR 500 suppressed significantly mRNA expression of IL-1$\beta$, IL-6 and TNF-$\alpha$. In serum, CR 500 decreased the amount of IL-1$\beta$, 12h, 24h and 48h respectively (p < 0.05), also decreased that of IL-6 and TNF-$\alpha$, 12h respectively (p < 0.05) after infarction. The more astrocytes were observed and neural plasticity was facilitated in the rat brain of CR 500 than that of sham control in immunohistochemistry. Conclusions : This results suggest that CR decrease infarction volume and improve functional motor recovery in acute stage in photothrombotic ischemic infarction model in the mechanism of anti-inflammation and promoting neural plasticity.

• Journal of Life Science
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• v.33 no.5
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• pp.414-421
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• 2023

Cnidium japonicum (C. japonicum) is a type of halophyte that inhabits soil of a high salinity, and according to previous studies, it is known to have antitumor effects. However, the skin's protective effect, particularly against UVB irradiation, has not been revealed. In this study, C. japonicum crude extract was studied to determine its effect on damage to human keratinocytes (HaCaT) induced by UVB irradiation, and ROS assays were performed, the results of which showed that C. japonicum crude extract affects UVB-induced photoaging damage in human keratinocytes. To examine inhibitory effects against the expressions of MMPs, RT-PCR and Western blot assay were performed by treating the crude extract at concentrations of 10, 50, and 100 ㎍/ml by irradiating UVB at 15 mJ/cm². As a result, it was confirmed that the mRNA and protein expression levels of MMP-1, MMP-3, and MMP-9 decreased in the group treated with C. japonicum crude extract, which also effectively regulated the antioxidant defense mechanism pathway by activating JNK, ERK, and p38. In conclusion, the current study suggested the possibility that C. japonicum could be used as a raw material for anti-photoaging cosmeceuticals in the future.

• The Korean Journal of Mycology
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• v.50 no.4
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• pp.263-274
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• 2022

Lentinula edodes is an important commercial mushroom and there have been several reports of viral infections in L. edodes. Two mycoviruses (LeV-HKB and LeNSRV1) were detected in Sanbaekhyang (NIFoS 2778) and Taehyanggo (NIFoS 4317), the sawdust-cultivated commercial strains. The vertical transmission rates of the viruses were investigated by detecting the viruses in 80 monokaryotic strains derived from basidiospores isolated from the fruiting bodies of each strain. Most of the monokaryotic strains were infected with the virus and the two viruses showed different levels of meiotic stability, with LeV-HKB showing higher meiotic stability than LeNSRV1. Therefore, it seems that the vertical transmission mechanism of mycoviruses is different depending on the virus species. We also examined the mycelial growth rate of the monokaryotic strains and compared the growth rate according to virus infection status. Although there was no statistically significant correlation between viral infection and mycelial growth rate, we found that the average growth rate was reduced by additional virus infection. We expect our data to contribute to a greater understanding of the mechanism of the vertical transmission of mycoviruses, and promoting breeding using virus-free monokaryotic strains.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.290-290
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• 2022

Powdery mildew is known to be one of the serious diseases in C. moschata cultivation. Plants infected with powdery mildew cause damage to cultivation areas such as occurrence of deformity fruit and decrease in quantity. also, it has been reported that many farms have difficulties in controlling powdery mildew due to the outbreak under various conditions throughout the year. Therefore, this study intends to perform a phenotype test and a genotype test for C. moschata 60 lines grown in Jenong S&T. Podospareaxanthii, known as a pathogen that causes powder mildew disease in pumpkins in Korea, was collected and used as an inoculation source, phenotype test was performed by examining the infection area rate(%) of powdery mildew disease that occurred in leaves 25 days after inoculation. It was determined that 0% of the infection area rate was in the first stage, 1 to 5% in the second stage, 6 to 15% in the third stage, 16 to 30% in the fourth stage, and 31% or more in the fifth stage, The first and second stages were judged as resistance, the third as moderate resistance, and the fourth and fifth stages as sensitivity. As a result of the phenotype test, it was confirmed that the resistance was 21 points, moderate resistance was 14 points, and sensitivity was 25 points. After searching for the genes related to powdery mildew resistance resistance, pm-0, CmbHLH87, and LOC111453072, 21 points of resistance and 9 points of moderate resistance identified through phenotype tests were identified through gel electrophoresis after polymerase chain reaction(PCR) using 5 primers related to 3 genes. As a result of genotype testing of a total 30 points, the CmbHLH87 and LOC111453072 gene were found to be resistant bands in all points, PMR1 was identified as 20 points for resistance, 4 points for moderate resistance, and 6 points for sensitivity, PMR2 was not identified in the entire band, and PMR5 was identified as 18 point for resistance, 3 points for moderate resistance, and 9 points for sensitivity. As a result, when comparing the phenotype test results and genotype test results, CmbHLH87 and LOC111453072 genes was 100% consistent in resistance and moderate resistance, PMR1 was 95.2% in resistance, 44.4% in moderate resistance, and PMR5 was 90% in resistance and 33.3% in moderate resistance, PMR2 was not consistent in resistance and moderate resistance. Therefore, it is expected that more accurate PMR test will be possible by using molecular markers(PMR1, PMR5) and by developing CmbHLH87 and LOC111453072 gene-related molecular markers.

• Journal of Ginseng Research
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• v.47 no.6
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• pp.755-765
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• 2023

Background: Caveolin-1, the scaffolding protein of cholesterol-rich invaginations, plays an important role in store-operated Ca²⁺ influx and its phosphorylation at Tyr14 (p-caveolin-1) is vital to mobilize protection against myocardial ischemia (MI) injury. SOCE, comprising STIM1, ORAI1 and TRPC1, contributes to intracellular Ca²⁺ ([Ca²⁺]_i) accumulation in cardiomyocytes. The purified extract of steamed Panax ginseng (EPG) attenuated [Ca²⁺]_i overload against MI injury. Thus, the aim of this study was to investigate the possibility of EPG affecting p-caveolin-1 to further mediate SOCE/[Ca²⁺]_i against MI injury in neonatal rat cardiomyocytes and a rat model. Methods: PP2, an inhibitor of p-caveolin-1, was used. Cell viability, [Ca²⁺]_i concentration were analyzed in cardiomyocytes. In rats, myocardial infarct size, pathological damages, apoptosis and cardiac fibrosis were evaluated, p-caveolin-1 and STIM1 were detected by immunofluorescence, and the levels of caveolin-1, STIM1, ORAI1 and TRPC1 were determined by RT-PCR and Western blot. And, release of LDH, cTnI and BNP was measured. Results: EPG, ginsenosides accounting for 57.96%, suppressed release of LDH, cTnI and BNP, and protected cardiomyocytes by inhibiting Ca²⁺ influx. And, EPG significantly relieved myocardial infarct size, cardiac apoptosis, fibrosis, and ultrastructure abnormality. Moreover, EPG negatively regulated SOCE via increasing p-caveolin-1 protein, decreasing ORAI1 mRNA and protein levels of ORAI1, TRPC1 and STIM1. More importantly, inhibition of the p-caveolin-1 significantly suppressed all of the above cardioprotection of EPG. Conclusions: Caveolin-1 phosphorylation is involved in the protective effects of EPG against MI injury via increasing p-caveolin-1 to negatively regulate SOCE/[Ca²⁺]_i.