• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.465-472
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• 2019

Candida albicans is an opportunistic human pathogen that causes infections. Candidiasis is often related to antifungal resistance because the pathogen has the ability to form biofilms. In a previous study, we found that the Salvia miltiorriza ethanol extract demonstrated anticandidal activity by altering membrane permeability and inhibiting the cell wall synthesis in C. albicans. Our results here demonstrate that $78{\mu}g/ml$ of the S. miltiorriza extract significantly diminished the early stage biofilms formed by 10 clinical C. albicans isolates by 51.3%; this was analyzed by 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) reduction assay. The effect of the S. miltiorrhiza extract on the adhesion of C. albicans cells to polystyrene plates and germ tube formation was examined via microscopic investigation. Although the density of the adhered cells was remarkably reduced up on incubation with $39{\mu}g/ml$ S. miltiorrhiza extract, germ tube formation by C. albicans was rarely affected. Quantitative real-time PCR analysis showed that the S. miltiorrhiza extract downregulated the expression of C. albicans hypha-specific genes, EAP1 by 34.7% (p < 0.001), ALS1 by 45.0% (p < 0.001), ALS3 by 48.1% (p < 0.001), and ECE1 by 21.3% (p = 0.006), respectively. Our data suggest that the S. miltiorrhiza ethanol extract significantly inhibited the early stage of biofilm formation by C. albicans by interfering with cell adhesion, by downregulating EAP1, ALS1 and ALS3, and presumably by modifying the cell wall and membrane structure.

• Journal of Life Science
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• v.29 no.10
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• pp.1144-1151
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• 2019

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease associated with various metabolic syndromes, such as obesity, dyslipidemia, hypertension, and diabetes. Cudrania tricuspidata is a medicinal plant distributed widely in Asia and has been used in clinical practice to treat various diseases. The aim of this study is to determine the lipid-lowering effects of C. tricuspidata fruit extract (CTE) using a cell model induced by free fatty acids (FFAs). HepG2 cells were exposed to 1mM FFAs (palmitic acid:oleic acid = 2:1) for 24 hr to simulate the conditions of NAFLD in vitro. CTE attenuated the increases of lipid accumulation, intracellular triglyceride, and cholesterol content and inhibited 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) activity in the HepG2 cells in a dose-dependent manner. Also, CTE inhibited the protein expression of lipogenesis-related genes, such as sterol regulatory element-binding protein-1/-2 (SREBP-1/-2), fatty acid synthase (FAS), and stearoyl CoA desaturase-1 (SCD-1) in FFAs-induced lipid accumulation in HepG2 cells. In addition, CTE-induced adenosine monophosphate-activated protein kinase (AMPK) phosphorylation in HepG2 cells. These results suggest that CTE attenuates hepatic lipid accumulation by inhibiting lipogenesis through the modulation of the AMPK signaling pathway on FFAs-induced lipogenesis in HepG2 cells and may potentially prevent NAFLD.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.3
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• pp.316-322
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• 2019

Helicobacter pylori, a gram-negative bacterium, is one of the risk factors that induces gastritis and gastric cancer. Therefore, much attention has been paid to the compounds that inhibit bacterial growth or eradicate bacteria. Evodiae fructus (EF), the fruit of Evodia rutaecarpa, has been used for treating diarrhea and abdominal pain. EF extract was already found to inhibit the growth of H. pylori. However, to the best of our knowledge, the effect of EF on the virulence factors of H. pylori has not been reported. In this study, when comparing the minimum inhibitory concentration (MIC) of the different methanol concentration extracts, the 95% methanol extract (EF95) showed the lowest MIC value. EF95 extract suppressed the expressions of cagA, vacA and ureB, but interestingly, it up-regulated the expression of ureA. A decrease in production of ammonia in the culture medium and the cell lysates indicated that EF95 inhibited the urease activity in H. pylori, which was the result of EF95 inhibiting the ureB expression. Although the mechanism by which EF95 extract regulates the virulence factors in H. pylori needs further study, EF95 could be used for treatment of gastric troubles induced by H. pylori.

• Journal of Life Science
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• v.29 no.1
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• pp.90-96
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• 2019

The objective of this study was to evaluate the potential of Diospyros malabarica stem extract, a natural materials, in oral health material. With this aim in mind, thin layer chromatography (TLC), TLC-bioautography, high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), scanning electron microscopy (SEM), and real-time qPCR were performed. The antibacterial activity of D. malabarica stem extract against Streptococcus mutans KCTC3065 was confirmed in an n-hexane fraction with low polarity. The molecular weight of the antibacterial compound was estimated to be 188 by ESI-MS analysis. The inhibitory effects of the extract on biofilm formation and gene expression related to biofilm formation of S. mutans were determined by SEM and real-time PCR analysis. The extract inhibited the formation of S. mutans biofilms at D. malabarica stem extract concentrations of 1 mg/ml, as shown by SEM. The real-time PCR analysis showed that the expression of the gtfC gene, which is associated with biofilm formation, was significantly decreased in a dose-dependent manner. Based on the above results, it can be concluded that D. malabarica stem extracts, a natural materials, can be used in oral health products to suppress the formation of biofilms by inhibiting tooth adhesion of S. mutans, a causative agent of dental caries.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.92-92
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• 2019

Excessive or chronic inflammation contributes to the pathogenesis of many inflammatory diseases such as sepsis, rheumatoid arthritis, and ulcerative colitis. Hibiscus syriacus L. has been used as a medicinal plant in many Asian countries, even though its anti-inflammatory activity has been unclear. Therefore, we investigated the anti-inflammatory effect of anthocyanin fractions from the H. syriacus L. varieties Pulsae (PS) on the lipopolysaccharide (LPS)-induced expression of proinflammatory mediators and cytokines in RAW264.7 macrophages. PS suppressed LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) secretion concomitant with downregulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Furthermore, PS inhibited the production of proinflammatory cytokines such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and IL-12 in LPS-stimulated RAW264.7 macrophages. Further study showed that PS significantly decreased LPS-induced nuclear translocation of the nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) subunits, p65 and p50. Molecular docking data showed that many anthocyanins from PS fit into the hydrophobic pocket of MD2 and bound to Toll-like receptor 4 (TLR4), indicating that PS inhibits the TLR4-MD2-mediated inflammatory signaling pathway. Especially, apigenin-7-O-glucoside most powerfully bound to MD2 and TLR4 through LYS122, LYS122, and SER127 at a distance of $2.205{\AA}$, $3.098{\AA}$, and $2.844{\AA}$ and SER441 at a distance of $2.873{\AA}$ (docking score: -8.4) through hydrogen bonding, respectively. Additionally, PS inhibited LPS-induced TLR4 dimerization/expression on the cell surface, which consequently decreased MyD88 recruitment and IRAK4 phosphorylation. PS completely blocked LPS-mediated mortality in zebrafish larvae by diminishing the recruitment of neutrophil and macrophages accompanied by low levels of proinflammatory cytokines. Taken together, our results indicate that PS attenuates LPS-mediated inflammation in both in vitro and in vivo by blocking the TLR4/MD2-MyD88/IRAK4-$NF-{\kappa}B$ axis. Therefore, PS might be used as a novel modulatory candidate for effective treatment of LPS-mediated inflammatory diseases.

• Korean Journal of Plant Resources
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• v.34 no.1
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• pp.31-36
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• 2021

Berchemia berchemiaefolia (Makino) Koidz. is found not only in korea, but also in japan and is known as a rare plant all over the world. In this study, we evaluated anti-inflammatory effect of B. berchemiaefolia (Makino) Koidz. leaves (BBK-L) in LPS-stimulated RAW264.7 cells. BBK-L inhibited the generation of NO through the suppression of iNOS experession. BBK-L attenuated the expression of iNOS, COX-2, TNF-α, IL-1β, IL-6 induced by LPS. BBK-L decreased LPS-mediated IκB-α degradation inhibite which resulted in the inhibition of NF-κB activation in RAW264.7 cells. In addition, BBK-L suppressed ERK1/2, JNK phosphorylation induced by LPS. BBK-L showed anti-inflammatory effect through blocking the generation of the inflammatory mediators such as NO, iNOS, COX-2, TNF-α, IL-1β, IL-6 via the inhibiting of NF-κB and MAPK signaling activation. These results suggests that BBK-L may have great potential for the development of anti-inflammatory drug to treat acute and chronic inflammatory disorders.

• Korean Journal of Clinical Laboratory Science
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• v.54 no.1
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• pp.9-14
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• 2022

Ulcerative colitis is a disease that causes inflammation in the mucosal or submucosal layer of the colon. Previous studies have reported that obesity increases the prevalence of ulcerative colitis and aggravates the progression. This study was therefore undertaken to investigate whether curcumin inhibits the progression of ulcerative colitis caused by obesity. Mice were bred on a high-fat diet to induce obesity, and curcumin was administered with the high-fat diet to confirm the anti-inflammatory effect. To induce ulcerative colitis, dextran sulfate sodium (DSS) was administered orally, and clinical symptoms of colitis were subsequently observed. For histological evaluation of curcumin, the colon, liver and abdominal fat tissue samples were prepared and analyzed by hematoxylin and eosin (H&E) and Alcian blue-periodic acid-Schiff (PAS) staining. Our results confirm that consumption of curcumin resulted in decreasing the score of the disease activity index, and inhibited shortening of the colon length. In addition, inflammatory cell infiltration and mucosal damage were inhibited in the colon tissue of ulcerative colitis exacerbated by obesity. We further confirmed that exposure to curcumin significantly reduced the steatosis area of the liver and adipocytes of abdominal fat. In conclusion, we believe that curcumin can be applied as a therapeutic agent to treat ulcerative colitis, by inhibiting the progression of colitis caused by obesity.

• Journal of Nutrition and Health
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• v.55 no.1
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• pp.59-69
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• 2022

Purpose: Natural medicinal plant extracts have recently attracted attention as health beneficial foods and potential therapeutic agents for prevention of various diseases. This study was undertaken to measure the anti-inflammatory effect of the ethanol-water fraction obtained from the above-ground portion of Spiraea prunifolia var. simpliciflora, a wild-growing plant in Korea. The final fraction used in this study was the H₂O-EtOH (40:60) fraction (SP60), which had the highest antioxidant activity, as determined in previous studies. Methods: The amounts of nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1β production were measured in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells exposed to SP60. Western blot was performed to measure the expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, and the activation of nuclear factor (NF)-κB. Results: SP60 exerted no cytotoxicity up to concentrations of 125 ㎍/mL. The levels of inflammatory cytokines, such as NO, TNF-α, IL-6, and IL-1β, were significantly decreased in LPS-stimulated RAW264.7 cells exposed to SP60. In addition, the expression levels of iNOS, COX-2, and phosphorylated p65 showed a concentration-dependent decrease subsequent to SP60 treatment. These results indicate that SP60 inhibits the LPS-induced production of inflammatory cytokines, iNOS, and COX-2, by inhibiting the activation of NF-κB, which is responsible for the expression of inflammatory mediators. Conclusion: The results presented in this study indicate that the H₂O-EtOH (40:60) fraction (SP60) extracted from the above-ground portion of Spiraea prunifolia var. simpliciflora has the potential to be developed as a medicine or healthcare food and functional material possessing anti-inflammatory effects. However, it is necessary to first confirm the anti-inflammatory effects of SP60 in in vivo models.

• The Korea Journal of Herbology
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• v.37 no.4
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• pp.31-38
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• 2022

Objectives : In this study, we investigated the synergistic protective effects of medicinal herbal mixture (HME) including Mori Ramulus (MR), Acanthopanacis Cortex (AC), Eucommiae Cortex (EC), and Black soybean (BS) in C2C12 cells, mouse myoblasts. Methods : Effects of HME on cell viability of C2C12 myoblasts were monitored by MTT assay. Anti-atrophic activity of HME was determined in myoblasts and myotubes under oxidative stress by H₂O₂. C2C12 myoblasts were differentiated into myotubes in a medium containing 2% horse serum for 6 days. After that, we measured that expression of MyoD and myogenine, the myogenic regulatory factors, to identify the mechanism of inhibiting muscle atophy after HME treatment. In addition, suppression of phosphorylation of Akt, FoxO3a and MARF-1, transcription factors of degradation proteins were analyzed via western blotting. Results : As a result of MTT, HME there was no show cytotoxicity up to a concentration of 1 mg/ml. The cytoprotective effects on oxidative stressed myoblast and myotube was better in HME extract than those of MR, AC, EU, and BS, respectively. HME treatment in Myotube induced by oxidative stress after H₂O₂ treatment increased Myo D, Myogenine activation, and Akt, FoxO3a phosphorylation and decreased expression of MuRF-1. As the results, HME has synergistic effects on protection against proteolysis of C2C12 myotubes through activation of the Akt signaling pathway under oxidative stress. Conclusions : These results suggest that HME may also be useful as a preventing and treating material for skeletal muscle atrophy caused by age-related diseases.

• Korean Journal of Poultry Science
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• v.49 no.2
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• pp.69-77
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• 2022

Antimicrobial peptides (AMPs) play an important role in innate immunity against pathogenic infections. AMPs exterminate pathogenic bacteria by disrupting cell membranes or inhibiting intracellular molecules. NK-2, first identified in pigs and derived from NK-lysin, has antimicrobial effects against bacteria and parasites. In this study, chimeric peptides (cpNK) of chicken and pig NK-2 and cpNK-derived peptides (cpNK-a1 and cpNK-a2) were synthesized, and their antimicrobial effects against various pathogenic bacteria such as Escherichia coli, Salmonella spp., Listeria monocytogenes, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus (MRSA) were investigated. The structure of chimeric peptides from chicken and pig NK-2, cpNK, include α-helix like NK-2 and peptide net charge was +9 like porcine NK-2. The cpNK peptide showed powerful bactericidal effects against most bacterial species, including MRSA, especially against gram-negative bacteria. Furthermore, cpNK-derived short peptides, cpNK-a1 and a2 also showed bactericidal activity, but the effects were weaker than those of cpNK. Therefore, we conclude that cpNK- and cpNK-derived short peptides have the potential to be used as antibiotic alternatives.