• 대한수의학회지
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• 제38권4호
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• pp.730-736
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• 1998

To elucidate whether or not innervation of purinergic nerve and functional action of $P_{2X}$-, $P_{2Y}$-purinoceptor, the neurogenic effects of perivascular nerve stimulation were investigated using by physiograph recording system in isolated coronary artery of pig. 1. The contractile responses induced by perivascular nerve stimulation (20V, 0.5msec, 20sec) were the frequency (2~64Hz) dependent manner. 2. The neurogenic contractile responses induced by perivascular nerve stimulation were inhibited by the pretreatment with either ATP or adenosine ($10^{-7}{\sim}10^{-4}M$). 3. The neurogenic contractile responses induced by perivascular nerve stimulation (20V, 16Hz, 0.5msec, 20sec) were increased by the pretreatment with reactive blue 2, but were not affected by the pretreatment with 8-phenyltheopylline ($10^{-5}M$). 4. The neurogenic contractile responses induced by perivascular nerve stimulation (20V, 16Hz, 0.5msec, 20sec) were inhibited by the desensitization of the P2X-purinoceptor using by treatment of $10^{-5}M$, -methylene ATP as 3 times over again. The accomplished present study on isolated coronary artery of pig suggest that purinergic nerve is innervated and that the neurogenic contractile response was mediated by activation of $P_{2X}$-purinoceptor and the neurogenic relaxative response was mediated by activation of both $P_1$ and $P_{2Y}$-purinoceptor.

• 대한수의학회지
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• 제39권4호
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• pp.702-709
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• 1999

The present study was performed to elucidate the effects of extracellular $Ca^{2+}$ on contractile responses in isolated porcine coronary artery ring using by perivascular nerve stimulation (PNS). Especially, the study was focused on the source of $Ca^{2+}$ on $P_{2X}$-purinoceptor mediated muscle contraction which one of $P_2$-purinoceptor subtypes. The following results can be drawn from these studies : 1. The phasic contractions induced by PNS were inhibited with muscarinic receptor antagonist, atropine ($10^{-6}M$). 2. The phasic contractions induced by PNS were significantly inhibited by sequential treatment with atropine and adrenergic neural blocker, guanethidine ($10^{-6}M$). 3. The phasic contractions induced by PNS were inhibited with $P_{2X}$-purinoceptor desensitization by repetitive application of $\alpha$,$\beta$-Me ATP ($10^{-4}M$). 4. The phasic contractions induced by PNS were so weakened in calcium-free medium. 5. The phasic contractions induced by PNS were inhibited with calcium channel blocker, verapamil ($10^{-6}{\sim}5{\times}10^{-6}M$). 6. The phasic contractions induced by PNS on pretreated with verapamil ($10^{-6}{\sim}5{\times}10^{-6}M$) were not changed by $\alpha$,$\beta$-Me ATP ($10^{-4}M$). These results demonstrate that the neurogenic phasic contractions induced by PNS are due to adrenergic-, cholinergic- and $P_{2X}$-purinergic receptors and the origin of $Ca^{2+}$ on $P_{2X}$-purinoceptor mediated muscle contraction is extracellular $Ca^{2+}$ through plasmalemmal $Ca^{2+}$ channels.

• 대한수의학회지
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• 제37권1호
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• pp.103-110
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• 1997

The experiments were carried out to elucidate the relationships between neurogenic effects of electrical transmural nerve stimulation and effect of adenosine 5'-triphosphate(ATP) to purinoceptor on the urinary bladder smooth muscle of pig. The results were as follows : 1. The contractile responses induced by electrical transmural nerve stimulation(10V or 20V, 0.5msec, 10sec) were the frequency(2~64Hz) dependent manner. 2. The contractile response induced by carbachol was responsed with a dose-dependent manner and the maximum contractility was $10^{-4}M$. 3. The contractile responses induced by ATP were increased in a dose-dependent manner ($10^{-5}{\sim}10^{-3}M$). 4. The contractile response induced by electrical transmural nerve stimulation(10V, 2~32Hz, 0.5msec, 10sec) was partially blocked by the treatment with atropine($10^{-5}M$), and was powerfully inhibited by 3 times of addition with ATP($10^{-5}M$). 5. The contractile response induced by electrical transmural nerve stimulation(10V, 2~32Hz, 0.5msec, 10sec) was partially blocked by the treatment with atropine($10^{-5}M$), and was completely blocked by the desensitization of the $P_{2X}$-purinoceptor using ${\alpha}$, ${\beta}$-methylene ATP($5{\times}10^{-5}M$). These results suggest that purinergic nerve was innervated, and ATP and acetylcholine was released by the electrical transmural nerve stimulation in urinary bladder smooth muscle of pig.

• Archives of Pharmacal Research
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• 제24권6호
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• pp.552-556
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• 2001

Histamine and arachidonic acid (AA) release was measured using the P2-purinoceptor antaongists, phospholipase $A_2{\;}(PLA_2)$ and cyclooxygenase (COX)/lipoxygenase (LOX) inhibitors to determine whether or not ATP-induced histamine release is associated with arachidonic acid (AA) release in rat peritoneal mast cells. ATP increased histamine release in a dose dependent manner, whereas adenosine did not. PPADS (a selective P2X-purinoceptor antagonist) and suramin (a nonselective P2X,2Y-purinoceptor antagonist) inhibited ATP-induced histamine release in a dose dependent manner. However, RB-2 (a P2Y-purinoceptor antagonist) did not block ATP-induced histamine release. Manoalide and oleyloxyethyl phosphorylcholine (OPC), secretory PLA$_2$ inhibitors, also inhibited ATP-induced histamine release dose-dependently. Both COX inhibitors (ibuprofen and indomethacin) and LOX inhibitors (baicalein and caffeic acid) inhibited ATP-induced histamine in a dose dependent manner. ATP significantly increased [$^3H$]AA release by 54%. PPADS and suramin significantly inhibited ATP-induced [3H]Ph release by 81% and 39%, respectively. ATP-induced histamine release was significantly inhibited by a variety of protein kinase inhibitors, such as bisindolmaleimide, genistein, methyl 2,5-dihydroxycinnamate, W-7 and trifluoperazine. Overall, the results suggest that ATP-induced histamine release is in part related to the PLA2-mediated AA metabolism and P2X-purinoceptors.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2002년도 제9회 학술 발표회 프로그램과 논문초록
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• pp.45-45
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• 2002

Extracellular ATP regulates a wide range of cellular function including the growth of prostate gland. Purinoceptors (ATP receptors) are divided into P2X (ligand-gated ion channels) and P2Y (G-protein-coupled receptor) subfamilies. In the present study, we investigated the types of purinoceptors in rat prostate neuroendocrine (RPNE) cells using whole-cell patch clamp technique, intracellular $Ca^{2+}$ measurement and RT-PCR analysis.(omitted)d)

• 대한수의학회지
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• 제31권4호
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• pp.389-395
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• 1991

가토 적출 신동맥에 있어서 purinergic 신경 분포를 perivascular nerve stimulation을 통해서 확인하였다. 1. 가토 적출 신동맥은 perivascular nerve stimulation에 의해 수축성 반응을 나타내었다. 2. Perivascular nerve stimulation에 의한 수축현상은 자극의 크기와 자극의 빈도에 의존적이었으며, 내피세포의 유무에는 영향을 받지 않았다. 3. Perivascular nerve stimulation에 의한 수축현상은 prazosin$(1{\mu}M)$에 의해 강하게 억제되어졌으며, tetrodotoxin$(1{\mu}M)$ 또는 ${\alpha},{\beta}-methylene$ATP$(10{\mu}M)$에 의한 $P_{2x}-purinoceptor $ desensitization에 의해 완전히 수축현상이 사라졌다. 이와같은 결과는 가토 신동맥에는 purinergic신경이 분포되어 있으며, perivascular nerve stimulation에 의해 purinergic 신경전달물질이 norepinephrine과 함께 유리되어지는 것을 시사하고 있다.

• The Korean Journal of Physiology and Pharmacology
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• 제25권6호
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• pp.525-532
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• 2021

We have investigated the relative roles of α₁-adrenoceptors and purinoceptors in contractions to low and high frequency stimulation of the mouse vas deferens, in terms of the time course of responses. In separate experiments, isometric contractile responses were obtained to 10 pulses at 1 Hz and 40 pulses at 10 Hz. Responses to 1 Hz stimulation consisted of a series of discrete peaks. The α_1A-adrenoceptor antagonist RS100329 (10^-9M-10^-7M) significantly reduced the response to the first pulse, the α_1D-adrenoceptor antagonist BMY7378 (10^-7M-10^-6M) significantly reduced the response to the first two pulses, and the non-selective α₁-adrenoceptor antagonist prazosin (10^-8M) reduced the response to the first 4 pulses at 1 Hz. Responses to 10 Hz stimulation consisted of an early peak response and a maintained plateau response. RS100329 significantly reduced the peak response but did not significantly affect the plateau response. Prazosin, significantly reduced both the peak and plateau responses. The α_1A-adrenoceptor antagonist RS17053 in high concentrations reduced mainly the plateau response leaving a clear early peak response. The plateau response of contraction was almost abolished by the purinoceptor antagonist suramin. These results suggest that there is a relatively minor early α_1D-adrenoceptor and a larger early α_1A-adrenoceptor component to stimulationevoked contractions of mouse vas deferens, but the major α₁-adrenoceptor component is revealed by prazosin to be α_1B-adrenoceptor mediated. α_1B-Adrenoceptor activation probably facilitates contractions mediated by other α₁-adrenoceptors and by purinoceptors. These results suggest that combined non-selective α₁-adrenoceptor blockade, particularly α_1B-adrenoceptor blockade, in addition to P2X1-purinoceptor blockade is useful in reducing male fertility.

• 동의생리병리학회지
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• 제17권2호
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• pp.529-541
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• 2003

Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.