• Preventive Nutrition and Food Science
• /
• 제22권1호
• /
• pp.9-15
• /
• 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.

• The Korean Journal of Physiology and Pharmacology
• /
• 제23권5호
• /
• pp.345-356
• /
• 2019

Docosahexaenoic acid (DHA), an omega-3-fatty acid, modulates multiple cellular functions. In this study, we addressed the effects of DHA on human umbilical vein endothelial cell calcium transient and endothelial nitric oxide synthase (eNOS) phosphorylation under control and adenosine triphosphate (ATP, $100{\mu}M$) stimulated conditions. Cells were treated for 48 h with DHA concentrations from 3 to $50{\mu}M$. Calcium transient was measured using the fluorescent dye Fura-2-AM and eNOS phosphorylation was addressed by western blot. DHA dose-dependently reduced the ATP stimulated $Ca^{2+}$-transient. This effect was preserved in the presence of BAPTA (10 and $20{\mu}M$) which chelated the intracellular calcium, but eliminated after withdrawal of extracellular calcium, application of 2-aminoethoxy-diphenylborane ($75{\mu}M$) to inhibit store-operated calcium channel or thapsigargin ($2{\mu}M$) to delete calcium store. In addition, DHA ($12{\mu}M$) increased ser1177/thr495 phosphorylation of eNOS under baseline conditions but had no significant effect on this ratio under conditions of ATP stimulation. In conclusion, DHA dose-dependently inhibited the ATP-induced calcium transient, probably via store-operated calcium channels. Furthermore, DHA changed eNOS phosphorylation suggesting activation of the enzyme. Hence, DHA may shift the regulation of eNOS away from a $Ca^{2+}$ activated mode to a preferentially controlled phosphorylation mode.

• Journal of Microbiology
• /
• 제40권3호
• /
• pp.224-229
• /
• 2002

The effect of human cytomegalovirus (HCMV) on three human monocyte cell lines at different stages of differentiation was investigated. While the viability of HL-60 cells or U-937 cells was not significantly affected by HCMV infection, the viability of THP-1 cells was reduced. Acridine orange/ethidiurn bromide staining revealed that the reduction of THP-1 cell viability was due to increased apoptotic death following HCMV infection. Apoptosis in HL-60 cells was not affected by HCMV infection, and induction of apoptosis of U-937 cells by HCMV was intermediate between HL-60 and THP-1 cells. Since HL-60 cells are the least differentiated and THP-1 cells are the most differentiated, the induction of apoptosis of human monocytes appears to be related to the degree of cell differentiation. Flow cytometric and confocal microscopic studies using fluorescent calcium indicator Fluo-3 suggested a significant increase in intracellular free calcium concentration (［Ca$\^$2+/］i) in THP-1 cells undergoing apoptosis by HCMV infection. Again ［Ca$\^$2+/］i in HCMV-infected HL-60 cells was not critically altered, and that in HCMV-infected U-937 cells was intermediate between THP-1 cells and HL-60 cells. Calcium influx blockers such as verapamil and nifedipine partially reversed HCMV-induced apoptosis in THP-1 cells.

• Archives of Pharmacal Research
• /
• 제19권6호
• /
• pp.456-461
• /
• 1996

Vascular relaxation action of crude extracts of two kinds of Coptidis rhizoma (Coptis chinensis and Coptis japonica, Ranunculaceae) was investigated and compared with that of berberine and palmatine, active alkaloid components of these plants. The results show that total extracts, berberine, and palmatine induced a concentration-dependent vasodilatation of rat thoracic aorta contracted with phenylephrine (PE). Palmatine, unlike to berberine, did not inhibit contraction induced by KCI. In calcium-free media, not only berberine but also crude extracts inhibited calcium-induced contraction. Furthermore, pretreatment of crude extracts inhibited contraction induced by PE noncompetitively. In PE-induced contraction, berberine was 2.5 times more potent than Coptis chinensis in the relaxation of rat aorta in terms of $IC_{50}$ values. Analysis of the effects of crude extracts on the Emax and $IC_{50}(PE)IC_{50}(KCI)$ ratios provides information on selectivity and indicates that extracts exhibit greater inhibition of the contrac tile response induced by PE than by KCI. We concluded that crude extracts have .alpha.-adrenoceptor blocking action and possesses inhibitory effect on calcium influx, which may be at least in part responsible for the antihypertensive action.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
• /
• pp.97.2-97.2
• /
• 2003

(1R,9S)-(${\beta}$)-Hydrastine (HS) at 10-50 ${\mu}$M has been proven to have an inhibitory effect on dopamine biosynthesis in PC12 cells by the inhibition of tyrosine hydroxylase (TH) activity and TH gene expression. In the present study, therefore, the effects of HS on the basal and K$\^$+/-induced dopamine release, and Ca$\^$2+/ influx induced by high K$\^$+/ and caffeine in PC12 cells were investigated. The dopamine release by high K$\^$+/ (56 mM) was inhibited by co-incubation of 20 ${\mu}$M HS. Application of HS also significantly reduced the magnitude of the maintained Ca$\^$2+/ influx induced by K$\^$+/ depolarization. (omitted)

• 생명과학회지
• /
• 제16권3호
• /
• pp.433-441
• /
• 2006

폴리아민은 거의 모든 세포에 있어서 성장과 분화에 필수적인 물질이다. 이들 폴리아민의 기작은 상당히 복잡하고 다양하여 그 정확한 작용기전은 아직 확실하지가 않다. 본 논문에서는 폴리아민이 세포 증식을 유도 하기도 하지만 일정농도 이상에서는 오히려 세포사멸을 초래한다는 결과를 밝히고자 한다. 본 실험에 사용된 인간의 전립선 암세포(LNCaP cells)에 있어서, 폴리아민 가운데 putrescien은 세포 증식에 거의 영향을 미치지 않았으나 spermidine과 spermine의 경우 10 ${\mu}M$ 이하에서는 세포 증식을 촉진하였다. 그러나, 20 ${\mu}M$ 이상의 농도에서는 농도와 시간의존적으로 세포사별을 유도 하였다. 폴리아민 처리에 의한 세포사멸의 초기과정인 핵 응축과 염색질 condensation이 Hoechst와 PI 염색에서 뚜렷이 관찰되었다. 또한, 폴리아민 처리시 anti-apoptotic protein으로 알려진 Bcl-2 protein의 발현은 거의 완전히 억제된 반면, pro-apoptotic protein으로 알려진 Bax의 발현은 현저히 증대되었다. 본 연구의 결과에 따르면, 폴리아민에 의해서 유도되는 세포사멸은 세포 내 칼슘농도 변화에 의한 것으로 사료된다. 전립선 암세포에 있어서 폴리아민 처리시 시간과 농도 의존적으로 세포 내 칼슘농도가 증가되었다. 세포막을 통한 칼슘이동을 억제하는 nifedipine과 flufenamic acid 등의 억제제를 처리한 실험 결과 세포내 칼슘증가는 세포 내부의 저장소로부터 칼슘의 유출보다는 주로 세포막상에 있는 비선택적 칼슘통로를 통한 외부의 칼슘 유입에 의한 것으로 판단된다.

• 대한약리학회지
• /
• 제23권1호
• /
• pp.33-44
• /
• 1987

면역보체가 결합되어 있는 zymosan 또는 열로 응집된 IgG에 의하여 활성화된 호중구에서의 NADPH oxidase 의존적인 $O_{\overline{2}}$의 생성과 탐식작용은 칼슘의 흡수과정과 일치하였다. 활성화된 백혈구의 반응은 세포외 칼슘 농도에 따라 항진되었으며, 이는 칼슘의 킬레이트제인 EGTA나 EDTA에 의하여 유의하게 억제되었다. 활성화된 백혈구로부터 $O_{\overline{2}}$의 생성은 dantrolene과 chlorpromazine에 의하여 억제되었다. 칼슘 길항제인 bepredil, diltiazem, verapamil, nifedipine, nimodipine은 효과적으로 활성화된 백혈구의 칼슘흡수, $O_{\overline{2}}$ 생성 그리고 탐식 작용을 억제하였고 NADPH oxidase 활성도 또한 억제하였다. 그러므로, 칼슘 길항제는 칼슘 유입을 억제하거나 칼슘의 세포내 재분포 및 NADPH oxidase 반응계에 작용하여 활성화된 백혈구에서의 $O_{\overline{2}}$의 생성과 백혈구의 탐식작용을 억제할 것으로 시사되었다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제4권2호
• /
• pp.113-120
• /
• 2000

To investigate the mechanism of smooth muscle contraction induced by emptying of intracellular $Ca^{2+}$ stores, we measured isometric contraction and $^{45}Ca^{2+}$ influx. $CaCl_2$ increased $Ca^{2+}$ store emptying- induced contraction in dose-dependent manner, but phospholipase C activity was not affected by the $Ca^{2+}$ store emptying-induced contraction. The contraction was inhibited by voltage-dependent $Ca^{2+}$ channel antagonists dose dependently, but not by TMB-8 (intracellular $Ca^{2+}$ release blocker). Both PKC inhibitors (H-7 and staurosporine) and tyrosine kinase inhibitors (genistein and methyl 2,5-dihydroxycinnamic acid) significantly inhibited the contraction, but calmodulin antagonists (W-7 and trifluoperazine) had no inhibitory effect on the contraction. The combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were greater than that of each one alone. In $Ca^{2+}$ store-emptied condition, $^{45}Ca^{2+}$ influx was significantly inhibited by verapamil, H-7 or genistein but not by trifluoperazine. However combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were not observed. Therefore, this kinase pathway may modulate the sensitivity of contractile protein. These results suggest that contraction induced by emptying of intracellular $Ca^{2+}$ stores was mediated by influx of extracellular $Ca^{2+}$ through voltage-dependent $Ca^{2+}$ channel, also protein kinase C and/or tyrosine kinase pathway modulates the $Ca^{2+}$ sensitivity of contractile protein.

• 대한약리학회지
• /
• 제32권3호
• /
• pp.417-424
• /
• 1996

본 연구는 C5a에 의해 자극된 호중구에서 세포내 칼슘유리와 세포외부로부터 칼슘유입에 있어 protein kinase C와 protein tyrosine kinase의 관여 여부를 조사하였다. Protein kinase C 억제제인 staurosporine과 H-7은 C5a에 의해 자극된 호중구에서 세포내 칼슘유리를 억제하였으나, 세포막을 교차한 칼슘유입이나 세포내 칼슘농도 증가에 영향을 나타내지 않았다. C5a에 의한 세포내 칼슘유리와 칼슘유입은 protein tyrosine kinase 억제제인 genistein과 methyl-2,5-dihydroxycinnamate에 의해서 억제 되었다. ADP에 의해 야기된 세포내 칼슘농도의 증가는 genistein과 methyl-2,5-dihydroxycinnamate에 의해서 억제되었으나 staurosporine과 H-7의 영향은 받지 않았다. Genistein과 methyl-2,5-dihydroxy-cinnamate는 thapsigargin을 처리한 호중구에서 칼슘유입을 감소시켰으나 이에 대한 staurosporine 과 H-7의 효과는 나타나지 않았다. 호중구를 phorbol 12-myristate 13-acetate로 전처치하였을때 세포내 칼슘증가에 미치는 C5a의 자극 효과는 감소하였다. 이상의 결과로 부터 protein tyrosine kinase는 C5a에 의해 활성화된 호중구에서 세포내 칼슘유리와 세포막을 교차한 칼슘유입의 조절에 관여할 것으로 추정된다.

• 한국동물학회:학술대회논문집
• /
• 한국동물학회 1999년도 한국생물과학협회 학술발표대회
• /
• pp.176.1-176
• /
• 1999

No Abstract, See Full Text