• Biomolecules & Therapeutics
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• v.25 no.2
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• pp.105-111
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• 2017

It has been known that RA, one of major constituents of Perilla frutescens which has been used as a traditional folk remedy for sedation in oriental countries, shows the anxiolytic-like and sedative effects. This study was performed to know whether RA may enhance pentobarbital-induced sleep through ${\gamma}-aminobutyric$ acid $(GABA)_A-ergic$ systems in rodents. RA (0.5, 1.0 and 2.0 mg/kg, p.o.) reduced the locomotor activity in mice. RA decreased sleep latency and increased the total sleep time in pentobarbital (42 mg/kg, i.p.)-induced sleeping mice. RA also increased sleeping time and number of falling sleep mice after treatment with sub-hypnotic pentobarbital (28 mg/kg, i.p.). In electroencephalogram (EEG) recording, RA (2.0 mg/kg) not only decreased the counts of sleep/wake cycles and REM sleep, but also increased the total and NREM sleep in rats. The power density of NREM sleep showed the increase in ${\delta}-waves$ and the decrease in ${\alpha}-waves$. On the other hand, RA (0.1, 1.0 and $10{\mu}g/ml$) increased intracellular $Cl^-$ influx in the primary cultured hypothalamic cells of rats. RA (p.o.) increased the protein expression of glutamic acid decarboxylase ($GAD_{65/67}$) and $GABA_A$ receptors subunits except ${\beta}1$ subunit. In conclusion, RA augmented pentobarbital-induced sleeping behaviors through $GABA_A-ergic$ transmission. Thus, it is suggested that RA may be useful for the treatment of insomnia.

• Biomolecules & Therapeutics
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• v.32 no.1
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• pp.84-93
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• 2024

Rosmarinic acid (RA) is a phenolic ester that protects human keratinocytes against oxidative damage induced by ultraviolet B (UVB) exposure, however, the mechanisms underlying its effects remain unclear. This study aimed to elucidate the cell signaling mechanisms that regulate the antioxidant activity of RA and confirm its cyto-protective role. To explore the signaling mechanisms, we used the human keratinocyte cell line HaCaT and SKH1 hairless mouse skin. RA enhanced glutamate-cysteine ligase catalytic subunit (GCLC) and glutathione synthetase (GSS) expression in HaCaT cells in a dose- and time-dependent manner. Moreover, RA induced nuclear factor erythroid-2-related factor 2 (NRF2) nuclear translocation and activated the signaling kinases protein kinase B (AKT) and extracellular signal-regulated kinase (ERK). Treatment with the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002, the ERK inhibitor U0126, and small interfering RNA (siRNA) gene silencing suppressed RA-enhanced GCLC, GSS, and NRF2 expression, respectively. Cell viability tests showed that RA significantly prevented UVB-induced cell viability decrease, whereas the glutathione (GSH) inhibitors buthionine sulfoximine, LY294002, and U0126 significantly reduced this effect. Moreover, RA protected against DNA damage and protein carbonylation, lipid peroxidation, and apoptosis caused by UVB-induced oxidative stress in a concentration-dependent manner in SKH1 hairless mouse skin tissues. These results suggest that RA protects against UVB-induced oxidative damage by activating AKT and ERK signaling to regulate NRF2 signaling and enhance GSH biosynthesis. Thus, RA treatment may be a promising approach to protect the skin from UVB-induced oxidative damage.

• Journal of dental hygiene science
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• v.20 no.4
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• pp.221-229
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• 2020

Background: The purpose of this study was to investigate the anti-oral microbial activity and anti-inflammatory effects of rosmarinic acid (RA) in lipopolysaccharide (LPS)-stimulated MC3T3-E1 osteoblastic cells on a titanium (Ti) surface during osseointegration, and to confirm the possibility of using RA as a safe natural substance for the control of peri-implantitis (PI) in Ti-based dental implants. Methods: A disk diffusion test was conducted to confirm the antimicrobial activity of RA against oral microorganisms. In order to confirm the anti-inflammatory effects of RA, inflammatory conditions were induced with 100 ng/ml of LPS in MC3T3-E1 osteoblastic cells on the Ti surface treated with or without 14 ㎍/ml of RA. The production of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface was confirmed using an NO assay kit and PGE2 enzyme-linked immunosorbent assay kit. Reverse transcription polymerase chain reaction and western blot analysis were performed to confirm the expression of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in total RNA and protein. Results: RA showed weak antimicrobial effects against Streptococcus mutans and Escherichia coli, but no antimicrobial activity against the bacteria Aggregatibacter actinomycetemcomitans and the fungus Candida albicans. RA reduced the production of pro-inflammatory mediators, NO and PGE2, and proinflammatory cytokines, TNF-α and IL-1β, in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface at the protein and mRNA levels. Conclusion: RA not only has anti-oral microbial activity, but also anti-inflammatory effects in LPS-stimulated MC3T3-E1 osteoblasts on the Ti surface, therefore, it can be used as a safe functional substance derived from plants for the prevention and control of PI for successful Ti-based implants.

• Advances in Traditional Medicine
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• v.10 no.3
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• pp.173-183
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• 2010

Perilla frutescens var. acuta Kudo (PF) is a traditional Korean medicinal herb for allergic reaction regulation. In the present study, we investigated the effect of 30% ethanol extract powder of PF (EPPF) and rosmarinic acid (RA), the active compound of EPPF on various allergic reactions using in vivo and in vitro models. EPPF and RA significantly inhibited compound 48/48-induced systemic anaphylactic reaction and histamine release (P < 0.05). In addition, EPPF and RA significantly inhibited passive cutaneous anaphylaxis (PCA) in a dose-dependent manner (P < 0.05). These effects were stronger than those of disodium cromoglycate, the reference drug tested. EPPF and RA also significantly inhibited production of inflammatory cytokines, tumor necrosis factor-a interleukin (IL)-6, and vascular endothelial growth factor on the PCA reaction and phorbol 12-myristate 13-acetate and calcium ionophore A23187-stimulated human mast cell line, HMC-1 cells (P < 0.05). Moreover, EPPF and RA showed an inhibitory effect on lipopolysaccharide (LPS)-induced IL-4 production from whole spleen cells. Finally, EPPF and RA significantly decreased IL-4-dependent IgE production by LPS-stimulated whole spleen cells (P < 0.05). In conclusion, these results indicate that EPPF has potent anti-allergic activities.

• Analytical Science and Technology
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• v.32 no.3
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• pp.96-104
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• 2019

Agastache rugosa, known as Korean mint, is a medicinal plant with many beneficial health effects. In this study, a simple and reliable HPLC-UV method was proposed for the quantification of rosmarinic acid (RA) in the aqueous extracts of A. rugosa. RA was selected as a quantification marker due to its easiness in procurement and analysis. The developed method involved chromatographic separation on a $C_{18}$ column ($250{\times}4.6mm$, $5{\mu}m$) at room temperature. The mobile phase consisted of water and acetonitrile both containing 2 % acetic acid and was run at a flow rate of $1mL\;min^{-1}$. The method was validated for specificity, linearity, precision, and accuracy. It was specific to RA and linear in the range of $50-300{\mu}g\;mL^{-1}$ ($r^2=0.9994$). Intra-day, inter-day, and inter-analyst precisions were ${\leq}0.91%\;RSD$, ${\leq}1.40%\;RSD$, and 1.94 % RSD, respectively. Accuracy was 93.3-95.9 % (${\leq}1.21%\;RSD$). The method could be applied to three batches of bulk samples and three batches of lab scale samples, which were found to be $0.64({\pm}0.04)mg\;g^{-1}$ and $0.48({\pm}0.02)mg\;g^{-1}$ for the dried raw materials of A. rugosa. The results show that the proposed method can be used as a readily applicable method for QC of health functional foods containing the aqueous extracts of A. rugosa.

• Journal of Bio-Environment Control
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• v.31 no.4
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• pp.319-324
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• 2022

The objective of this study was to determine the proper night interruption of photoperiods and dark periods for accumulating bioactive compounds of Agastache rugosa without decreasing plant growth. Five-week-old seedlings were transplanted in a DFT system with white LEDs. A. rugosa was treated with night interruption time treatments of 18:1:2:3, 18:2:2:2, 18:3:2:1 (light:dark:light:dark), and 20:4 (control) for 4 weeks. There were no significant differences except for leaf length, leaf width, and the number of flowers. The content of antioxidants in the shoot of A. rugosa was high in tilianin and acacetin, and the content of rosmarinic acid (RA) was significantly higher in the underground part. The RA content per dry weight of A. rugosa was 47.92 and 51.46% higher than that of the control in 18:1:2:3 and 18:2:2:2, and tilianin and acactin per dry weight were significantly higher in 18:3:2:1. There was no significant difference in growth due to the same day light integral, but 18:2:2:2 showed high total polyphenol contents. Therefore, it is thought that the effect of increasing secondary metabolites of A. rugosa without degradation of growth can be expected through night interruption treatment in plant factory cultivation systems using artificial light.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.64-64
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• 2019

Screening and identification of genetic resources based on their phytoconstituents and morphological characters potentially provide baseline data for researchers, breeders, and nutraceutical companies who wish to formulate a nutrient-dense diet and health beneficial supplement. Thus, we evaluated the amount of total phenolic content and major phenolic compounds; examined if phenolic compounds could be used as distinguishing factors for perilla genetic resources; and investigated the relation between some quantitative and qualitative morphological characters with the contents of phenolic compounds in 360 accessions obtained from National Agrobiodiversity Center gene bank, Jeonju, Korea. Total phenolic content (TPC) was estimated using Folin-Ciocalteu colorimetric assay. Individual phenolic compounds were determined using an Ultra-High Performance Liquid Chromatography system equipped with Photodiode Array detector. Considerable variations were observed in TPC (7.99 to 117.47 mg GAE/g DE), rosmarinic acid (RA) (ND to 19.19 mg/g DE), caffeic acid (CA) (ND to 0.72 mg/g DE), apigenin-7-O-diglucuronide (ADG) (ND to 1.24 mg luteolin equivalent (LUE)/g DE), scutellarein-7-O-glucuronide (SG) (ND to 4.32 mg LUE/g DE), and apigenin-7-O-glucuronide (AG) (ND to 1.60 mg LUE/g DE). RA was the most dominant phenolic compound in most accessions (95.3%) followed by SG. The adaxial leaf color was light green, green and dark green in 13.8%, 65.0%, and 21.1 % of the accessions, respectively. 78.8% of the accessions had light green color at the abaxial side with the remaining being described as green. Most of the accessions (96.9%) were cordate shape, the remaining being eclipse. Intensities of green pigment at abaxial and adaxial leaf surfaces were correlated with contents of individual phenolic compounds and TPC whereas leaf length and width had no correlation with TPC, CA and RA, and negatively correlated with ADG, AG, and SG. Leaf shape was not related with content of phenolic compounds, color of leaves, or the length or width of leaves. Accessions IT57426, IT157434, IT267710, and IT267712 which contained relatively high contents of TPC and major phenolic compounds (RA and SG) could be used for further research in breeding and bioassay test. Our study result showed the contents of total phenolics and individual phenolic compounds along with the morphological characters could be useful distinguishing factors for perilla genetic resources.

• Journal of Plant Biotechnology
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• v.3 no.2
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• pp.107-112
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• 2001

The kinetics of Ti-transformed Salvia miltiorrhiza cell cultures was studied in 250-$m\ell$ shake flasks by using B5 medium with addition of 30 gfL of sucrose. In the cell cultures, the maximum cell mass obtained was 11.5 g DW/L on day 15. The highest amount of phenolic compounds - rosmarinic acid (RA) and lithospermic acid B (LAB) reached 871.3 mg/L (day 15) and 121.3 mg/L (day 13), respectively. The total tanshinone production, i.e., intracellular plus extracellular cryptotanshinone, tanshinone 1, and tanshinone IIA, was 5.3 mg/L on day 13. For the cultivations in 2.4-L stirred bioreactors, the residual sugar level and medium conductivity were a little higher in a small turbine impeller reactor ($T_s$) than those in a large turbine impeller reactor ($T_L$), while a higher cell density was obtained in the $T_L$. For the production of tanshinones and phenolics, better results were obtained in the $T_L$ than in the $T_s$. In the $T_L$, similar or even a little higher production titers of tanshinones and phenolic compounds were achieved compared to those in the flasks. The results suggest that the shake flask results could be successfully scaled up to the $T_L$ reactor. Such a large impeller reactor like $T_L$ may be better than a small impeller one for the large-scale production of the valuable metabolites by the suspension cultures of Ti transformed S.miltiorrhiza cells. This is considered due to the beneficial culture environment in the $T_L$, such as low shear rates as estimated theoretically.

• Restorative Dentistry and Endodontics
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• v.39 no.2
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• pp.95-103
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• 2014

Objectives: This study evaluated the effect of three antioxidizing agents on pullout bond strengths of dentin treated with sodium hypochlorite. Materials and Methods: Root canals of 75 single-rooted human teeth were prepared. Fifteen teeth were irrigated with normal saline for a negative control group, and the remaining 60 teeth (groups 2 - 5) with 2.5% NaOCl. The teeth in group 2 served as a positive control. Prior to post cementation, the root canals in groups 3 - 5 were irrigated with three antioxidizing agents including 10% rosmarinic acid (RA, Baridge essence), 10% hesperidin (HPN, Sigma), and 10% sodium ascorbate hydrogel (SA, AppliChem). Seventy-five spreaders (#55, taper .02, Produits Dentaires S.A) were coated with silica and silanized with the Rocatec system and ceramic bond. All the prepared spreaders were cemented with a self-adhesive resin cement (Bifix SE, Voco Gmbh) in the prepared canals. After storage in distilled water (24 h/$37^{\circ}C$), the spreaders were pulled out in a universal testing machine at a crosshead speed of 1.0 mm/min. Pull-out strength values were analyzed by one-way ANOVA and Tukey's HSD test (${\alpha}$ = 0.05). Results: There were significant differences between study groups (p = 0.016). The highest pullout strength was related to the SA group. The lowest strength was obtained in the positive control group. Conclusions: Irrigation with NaOCl during canal preparation decreased bond strength of resin cement to root dentin. Amongst the antioxidants tested, SA had superior results in reversing the diminishing effect of NaOCl irrigation on the bond strength to root dentin.

• Restorative Dentistry and Endodontics
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• v.39 no.4
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• pp.303-309
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• 2014

Objectives: Hydrogen peroxide ($H_2O_2$) surface treatment of fiber posts has been reported to increase bond strength of fiber posts to resin cements. However, residual oxygen radicals might jeopardize the bonding procedure. This study examined the effect of three antioxidant agents on the bond strength of fiber posts to conventional and self-adhesive resin cements. Materials and Methods: Post spaces were prepared in forty human maxillary second premolars. Posts were divided into five groups of 8 each: G1 (control), no pre-treatment; G2, 10% $H_2O_2$ pre-treatment; G3, G4 and G5. After $H_2O_2$ application, Hesperidin (HES), Sodium Ascorbate (SA) or Rosmarinic acid (RA) was applied on each group respectively. In each group four posts were cemented with Duo-Link conventional resin cement and the others with self-adhesive BisCem cement. Push-out test was performed and data were analyzed using 2-way ANOVA and tukey's post-hoc test (${\alpha}=0.05$). Results: There was a statistically significant interaction between the cement type and post surface treatment on push-out bond strength of fiber posts (p < 0.001, F = 16). Also it was shown that different posts' surface treatments significantly affect the push-out bond strength of fiber posts (p = 0.001). $H_2O_2$ treated posts (G2) and control posts (G1) cemented with Duo-link showed the highest ($15.96{\pm}5.07MPa$) and lowest bond strengths ($6.79{\pm}3.94$) respectively. Conclusions: It was concluded that $H_2O_2$ surface treatment might enhance the bond strength of fiber posts cemented with conventional resin cements. The effect of antioxidants as post's surface treatment agents depends on the characteristics of resin cements used for bonding procedure.