• The Korean Journal of Pharmacology
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• v.30 no.2
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• pp.243-254
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• 1994

Inflammatory diseases, allergic and asthmatic disorders are caused by the mediator release from the activation of the phospholipase C (PLC), phospholipase D (PLD), methyltransferase or adenylate cyclase etc. during IgG or IgE cross-linking of high affinity receptors on mast cells or basophil surface. One important enzyme activated after IgG or IgE receptor cross-linking is PLD, the enzyme which converts phosphatidylcholine (PC) to phosphatidic acid (PA). Under the hypothesis that these may be some differences in mediator release according to the difference in PLD activity, we attempted to confirm the ginseng saponin effects on the PLD activity. We examined the PLD activity during the passively sensitized mast cell activation in the presence of single component of ginsenosides $(Rc,\;Rg_1,\;Rg_2,\;Rg_3)$. We also measured the amount of mediators (histamine and leukotrienes) released by stimulating with ovalbumin (OA) or calcium ionophore (CaI), Guinea Pig lung mast cells were purified using enzyme digestion, count current elutriation, and discontinuous Percoll density gradient. In purified mast cells prelabeled with $[^3H]$ arachidonic acid or $[^3H]$ palmitic acid, PLD activity was assessed more directly by the production of labeled PEt by PLD-mediated transphosphatidylation in the presence of ethanol. Histanine release was determined by Spectrophotofluorometry, and leukotrienes by radioimmunoassay. The PLD activity during the passively sensitized mast cell activation is increased up to $3{\sim}5times$. The PLD activity during the passively sensitized mast cell activation in the presence of all ginsenosides is decreased up to $4{\sim}11$ times. $Rg_l\;and\;Rg_2$ ginsenoside pretreatment decreased histamine and leukotrienes by 50% in the OA-induced or by 40% in the Cal-induced mast cell after passively sensitization. Rc pretreatment poorly decreased histamine but leukotrienes decreased by 70% in the OA-induced or by 35% in the Cal-induced mast cell. $Rg_3$ ginsenoside pretreatment increased histamine release without challenging OA or Cal but leukotrienes decreased. These observations indicate that single unit of ginsenosldes may be an important contributor to inhibit the release of histamine and leukotrienes in the guinea pig lung mast cells, that inhibits the PLD-mediated formation of DAG evoked by mast cell activation.

• Korean Journal of Plant Tissue Culture
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• v.27 no.5
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• pp.375-377
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• 2000

It is now generally accepted that a phosphoinositide cycle is involved in the transduction of a variety of signals in plant cells. In animal cells, the hydrolysis of phosphatidyl-4,5-bisphosphate catalysed by phosphatidylinositol - specific phospholipase C yields to D-myo-inositol - 1,4,5-trisphosphate and diacylglycerol, which are well known second messengers. The binding of InsP$_3$to a receptor located on the endoplasmic reticulum triggers a calcium release from the endoplasmic reticulum. We have detected and partially characterised key components of phosphoinositide signaling. First, tobacco microsomal fraction and plasma membrane PI-PLC. Consecutively, using a radioligand binding assay we have identified a $Ca^{2+}$ -dependent high affinity InsP$_3$binding site in microsomal membrane fraction vesicle preparation and then we have measured inositol-1,4,5-trisphosphate induced calcium release from tobacco microsomal fraction. These findings suggest that phosphoinositide signaling system is present and operates in the tobacco suspension culture.e.

• Journal of Microbiology and Biotechnology
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• v.22 no.5
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• pp.622-627
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• 2012

Mast cells and basophils are important effector cells in immunoglobulin-E (IgE)-mediated allergic reactions. Using the human basophilic KU812F cells, we assessed the inhibitory effects of 6-methoxyluteolin, isolated from Chrysanthemum zawadskii, in the $Fc{\varepsilon}RI$-mediated allergic reaction. We determined that 6-methoxyluteolin inhibited anti-$Fc{\varepsilon}RI$ ${\alpha}$ chain antibody (CRA-1)-induced histamine release, as well as elevation of intracellular calcium concentration $[Ca^{2+}]_i$ in a dose-dependent manner. Moreover, the inhibitory effects of 6-methoxyluteolin on the cell surface expression and the mRNA level of the $Fc{\varepsilon}RI$ ${\alpha}$ chain were determined by flow cytometric analysis and reverse transcription-polymerase chain reaction (RT-PCR), respectively. Therefore, these results show that 6-methoxyluteolin is a potent inhibitor of histamine release and calcium influx via down-regulation of the $Fc{\varepsilon}RI$ ${\alpha}$ chain.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.47 no.2
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• pp.99-105
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• 2021

This study was conducted to identify the efficacy related to skin barrier functions such as anti-inflammatory, anti-allergic, cornified envelope formation and the alleviation effect of keratinocyte damage caused by S. aureus using the callus extract, which was induced and extracted from the leaves of Gynostemma pentaphyllum in Ulleungdo Island. In order to confirm the anti-inflammatory effect of the G. pentaphyllum callus extract on the skin, the expression of inflammatory cytokines was investigated in primary epidermal keratinocytes (HEKa) activated with PAR-2 agonist, and the G. pentaphyllum callus extract showed IL-8, IL-25, and TSLP production inhibitory effect. β-Hexosaminidase assay using RBL-2H3 cells was performed to confirm the anti-allergic efficacy, and G. pentaphyllum callus extract showed the effect of inhibiting the release of β-hexosaminidase. In addition, G. pentaphyllum callus extract showed the cornified envelope formation effect in HaCaT cells, and through the co-culture experiment with HaCaT cells and S. aureus, it showed alleviation effect of keratinocyte damage caused by S. aureus. Therefore, G. pentaphyllum callus extract is considered to be a useful cosmetic material for improving skin barrier with anti-inflammatory, anti-allergic and alleviation effect of keratinocyte damage caused by S. aureus.

• Korean Journal of Food Science and Technology
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• v.40 no.6
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• pp.702-706
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• 2008

Punica granatum (PG) evidences a variety of physiological properties, including anti-diabetic, anti-cancer, antiinflammatory, anti-microbial, and anti-oxidative activities. Using the human basophilic KU812F cells, the inhibitory effects of the methanolic extract of PG seed, shell, and juice on calcium ionophore, A23187-induced degranulation were assessed. All of the PG extracts inhibited A23187-induced intracellular $Ca^{2+}$ levels, ${\beta}$-hexosaminidase, and histamine release in a dose-dependent manner. These results showed that all of the PG extracts are potent inhibitors of degranulation in allergic reactions, via the suppression of $Ca^{2+}$ influx.

• YAKHAK HOEJI
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• v.41 no.4
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• pp.399-406
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• 1997

The effects of different $K^+$ channel blockers were investigated on the non-adrenergic non-cholinergic (NANC) relaxations in the circular muscle of the rabbit proximal stomach. Non-selective blockers of $K^+$ channels, 4-aminopyridine (4-AP, 3~30${\mu}M$) and tetraethylammonium (TEA, 100~1000${\mu}M$) significantly enhanced the NANC relaxations in a concentration-dependent manner. The enhancement was more prominent for the NANC relaxations induced by the electric field stimulation (EFS) with lower frequencies. Blockers of large conductance $Ca^{2+}$-activated $K^+$ channels, charybdotoxin and iberiotoxin, a blocker of small conduntance $Ca^{2+}$-activated $K^+$ channels, apamin and a blocker of ATP-sensitive $K^+$ channels, glibenclamide had no effect on the NANC relaxations, respectively. Exogeneous administration of nitric oxide (NO, 1~30${\mu}M$) caused concentration-dependent relaxations which showed a similarity to those obtained with EFS. None of the $K^+$ channel blockers had an effect on the concentration-dependent relaxation in response to NO. These results suggest that prejunctional $K^+$ channels regulate the release of NO from the NANC nerve in the rabbit proximal stomach as the inhibition of prejunctional $K^+$ channels increases the NANC relaxation induced by the EFS.

• The Journal of Korean Medicine
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• v.19 no.1
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• pp.251-270
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• 1998

The effects of Gamigunshimtang on the isolated perfused ischemic heart in rats, heart rates, left ventricular pressure, cardiac blood flow and cardiotoxicity were stu.died in H9C2 myoblast cell, myocardial slice culture The results were as follows: 1. The administration of Gamigunshimtang to the rat recovered effectively heart rate, left ventricular pressure and flow rate from the experimental ischemia in perfused rat heart. The release of lactic dehydrogenase after the ischemia also decreased compared to the control group. 2. The administration of Gamigunshimtang to H9C2 myoblast culture enhanced the cell proliferation and protected against doxorubicin and allylamine induced release of the lactic dehydrogenase into the culture medium. It also protected effectively against doxorubicin and allylamine induced decrease of Ca ATPase activity and the increase of NADPH-cytochrome C reductase activity in the microsome. 3. The administration of Gamigunshimtang to the rat myocardial slice culture protected effectively against doxorubicin and allylamine induced decreases of protein synthesis and ATP content, and increases of cvtosolic enzyme, creatin kinase into the medium and lipid peroxidation.

• Journal of Ginseng Research
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• v.45 no.4
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• pp.490-497
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• 2021

Background and objective: Synthetic ginsenoside compounds G-Rp (1,3, and 4) and natural ginsenosides in Panax ginseng 20(S)-Rg3, Rg6, F4 and Ro have inhibitory actions on human platelets. However, the inhibitory mechanism of ginsenoside Rk1 (G-Rk1) is still unclear thus, we initiated investigation of the anti-platelet mechanism by G-Rk1 from Panax ginseng. Methodology: Our study focused to investigate the action of G-Rk1 on agonist-stimulated human platelet aggregation, inhibition of platelet signaling molecules such as fibrinogen binding with integrin α_IIbβ₃ using flow cytometry, intracellular calcium mobilization, fibronectin adhesion, dense granule secretion, and thromboxane B2 secretion. Thrombin-induced clot retraction was also observed in human platelets. Key Results: Collagen, thrombin, and U46619-stimulated human platelet aggregation were dose-dependently inhibited by G-Rk1, while it demonstrated a more effective suppression on collagen-stimulated platelet aggregation using human platelets. Moreover, G-Rk1 suppressed collagen-induced elevation of Ca²⁺ release from endoplasmic reticulum, granule release, and α_IIbβ₃ activity without any cytotoxicity. Conclusions and implications: These results indicate that G-Rk1 possess strong anti-platelet effect, proposing a new drug candidate for treatment and prevention of platelet-mediated thrombosis in cardiovascular disease.

• International Journal of Oral Biology
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• v.44 no.2
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• pp.62-70
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• 2019

Xylitol is well-known to have an anti-caries effect by inhibiting the replication of cariogenic bacteria. In addition, xylitol enhances saliva secretion. However, the precise molecular mechanism of xylitol on saliva secretion is yet to be elucidated. Thus, in this study, we aimed to investigate the stimulatory effect of xylitol on saliva secretion and to further evaluate the involvement of xylitol in muscarinic type 3 receptor (M3R) signaling. For determining these effects, we measured the saliva flow rate following xylitol treatment in healthy individuals and patients with dry mouth. We further tested the effects of xylitol on M3R signaling in human salivary gland (HSG) cells using real-time quantitative reverse-transcriptase polymerase chain reaction, immunoblotting, and immunostaining. Xylitol candy significantly increased the salivary flow rate and intracellular calcium release in HSG cells via the M3R signaling pathway. In addition, the expressions of M3R and aquaporin 5 were induced by xylitol treatment. Lastly, we investigated the distribution of M3R and aquaporin 5 in HSG cells. Xylitol was found to activate M3R, thereby inducing increases in $Ca^{2+}$ concentration. Stimulation of the muscarinic receptor induced by xylitol activated the internalization of M3R and subsequent trafficking of aquaporin 5. Taken together, these findings suggest a molecular mechanism for secretory effects of xylitol on salivary epithelial cells.

• Preventive Nutrition and Food Science
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• v.22 no.1
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• pp.9-15
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• 2017

Vaccinium angustifolium, commonly known as the lowbush blueberry, is a rich source of flavonoids, with which various human physiological activities have been associated. The present study focuses on the investigation of the effect of the methanolic extract of V. angustifolium root extract (VAE) on high affinity immunoglobulin E receptor ($Fc{\varepsilon}RI$) ${\alpha}$ chain antibody (CRA-1)-induced allergic reaction in human basophilic KU812F cells. The total phenolic content of VAE was found to be $170{\pm}1.9mg$ gallic acid equivalents/g. Flow cytometry analysis revealed that the cell surface expression of $Fc{\varepsilon}RI$ was suppressed in a concentration-dependent manner upon culture with VAE. Reverse-transcriptase polymerase chain reaction analysis showed that the mRNA level of the $Fc{\varepsilon}RI$ ${\alpha}$ chain was reduced in a concentration-dependent manner as a result of VAE treatment. Western blot analysis revealed that the protein expression of $Fc{\varepsilon}RI$ and the phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 were concentration-dependently inhibited by VAE. We determined that VAE inhibited anti-CRA-1-induced histamine release, in addition to the elevation of intracellular calcium concentration ([$Ca^{2+}$]i), in a concentration-dependent manner. These results indicate that VAE may exert an anti-allergic effect via the inhibition of calcium influx and histamine release, which occurs as a result of the down-regulation of $Fc{\varepsilon}RI$ expression through inhibition of ERK 1/2 activation.