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Compound 48/80 activates mast cell phospholipase D via heterotrimeric GTp-binding proteins

JOURnAL OF pHARMACOLOGY AnD EXpERIMEnTAL THERApEUTICS 2000年第1期292卷 122-130页

作者： Chahdi, A Fraundorfer, pF Beaven, MA NHLBI Lab Mol Immunol NIH Bethesda MD 20892 USA

previous studies have indicated the presence of a cholera toxin-sensitive phospholipase D (pLD) in cultured RBL-2H3 mast cells that is synergistically activated via calcium, protein kinase C, and another unidentified signal. Here we identify a third potential signal for activation transduced by a pertussis toxin-sensitive trimeric GTp-binding protein, most likely via G(i2) or G(i3). Quercetin-treated RBL-2H3 cells in which expression of G(alpha i2) and G(alpha i3) is enhanced more than 7-fold respond to the G(i) stimulant compound 48/80 with the activation of pLD, a transient activation of phospholipase C, and enhanced membrane GTpase activity. The activation of pLD was blocked in pertussis toxin-treated cells and, as with other stimulants of pLD, was enhanced in cholera toxin-treated cells. The pLD response to compound 48/80 was only partially inhibited by calcium deprivation and inhibition of protein kinase C to indicate a component of the response that was independent of calcium, protein kinase C, and, presumably, phospholipase C. Based on these and other data, we hypothesized that beta gamma-subunits, released from G(i2) or G(i3) by compound 48/80 or from G(s) by cholera toxin, provide an additional signal for the activation of pLD. Consistent with this hypothesis, recombinant G(beta 2 gamma 2) subunits, but not G(alpha i-3) subunits, at concentrations of 50 to 300 nM markedly synergized pLD activation by compound 48/80 in permeabilized RBL-2H3 cells.

关键词：印迹法蛋白质钙/缺乏细胞膜/药物作用细胞膜/酶学细胞培养的霍乱毒素药物相互作用电泳 GTp磷酸水解酶类/代谢异源三聚体GTp结合蛋白质类/生理学肥大细胞/药物作用肥大细胞/代谢通透性百白咳毒素磷脂酶D/代谢蛋白激酶C/拮抗剂和抑制剂槲皮素/药理学信号传导时间因素毒力因子鲍特菌属 p-甲氧基-n-苯丙胺/药理学

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Gi-mediated activation of mitogen-activated protein kinase (MApK) pathway by receptor mimetic basic secretagogues of connective tissue-type mast cells: Bifurcation of arachidonic acid-induced release upstream of MApK

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JOURnAL OF pHARMACOLOGY AnD EXpERIMEnTAL THERApEUTICS 1999年第3期289卷 1654-1661页

作者： Shefler, I Seger, R Sagi-Eisenberg, R Tel Aviv Univ Sackler Fac Med Dept Cell Biol & Histol IL-69978 Tel Aviv Israel Weizmann Inst Sci Dept Regulat Biol IL-76100 Rehovot Israel

The family of basic secretagogues of connective tissue mast cells act as receptor mimetic agents, which trigger exocytosis by directly activating G proteins. We now demonstrate that pertussis toxin (ptx)-sensitive Gi proteins, activated by compound 48/80 (c48/80), a potent member of this family, also activate the p42/p44 MAp kinases (MApKs). This activation was potentiated by the protein tyrosine phosphatase inhibitor vanadate, whereas the tyrphostin AG-18, a competitive inhibitor of protein tyrosine kinases (pTKs);the protein kinase C inhibitors K252a and GF109203X;the phosphatidylinositol-3-kinase (pI-3K) inhibitors wortmannin and LY294002;and EGTA have abolished this activation. These results suggest that c48/80 activated the p42/p44 MApKs via a mechanism that involves pTKs, protein kinase C, phosphatidylinositol-3-kinase and Ca2+ as mediators. protein tyrosine phosphorylation and activation of the p42/p44 MApKs were closely correlated with stimulation of arachidonic acid (AA) release by c48/80 but not with histamine secretion. However, whereas pD98059, the inhibitor of the MApK kinase has abrogated MApK activation, this inhibitor failed to effect release of AA. We therefore conclude that by activating ptx-sensitive Gi protein(s), the basic secretagogues of mast cells stimulate multiple signaling pathways, which diverge to regulate the production and release of the different inflammatory mediators. Whereas the signaling pathway responsible for triggering histamine release is pTK independent, the pathway responsible for the stimulation of AA release bifurcates downstream to pTKs but upstream to the activation of MApKs.

关键词：雄二烯类/药理学花生四烯酸类/代谢卡唑类/药理学色酮类/药理学结缔组织细胞/细胞学结缔组织细胞/生理学依他酸/药理学酶激活酶抑制剂/药理学 GTp结合蛋白质α亚单位 Gi-Go/代谢组胺释放肌醇磷酸类/代谢动力学肥大细胞/细胞学肥大细胞/药物作用肥大细胞/生理学丝裂原活化蛋白激酶1/代谢丝裂原活化蛋白激酶3 丝裂原激活蛋白激酶类吗啉类/药理学百白咳毒素磷酸酪氨酸/代谢大鼠 Wistar 信号传导酪氨酸磷酸化抑制剂/药理学钒酸盐类/药理学毒力因子鲍特菌属/药理学 p-甲氧基-n-苯丙胺/药理学动物大鼠

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Inhibitory effect of mast cell-mediated immediate-type allergic reactions by sulfapyridine

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IMMUnOpHARMACOLOGY AnD IMMUnOTOXICOLOGY 2000年第2期22卷 253-266页

作者： Kim, HM An, nH Yi, BH Chae, HJ Kim, HR Moon, SJ Kim, JJ park, ST Baek, SH Wonkwang Univ Sch Med Coll Pharm Iksan 570749 Chonbuk South Korea Wonkwang Univ Sch Dent Dept Dent Pharmacol Iksan 570749 Chonbuk South Korea Wonkwang Univ Sch Med Coll Oriental Med Iksan 570749 Chonbuk South Korea Wonkwang Univ Sch Med Dept Anat Iksan 570749 Chonbuk South Korea Wonkwang Univ Ctr Oriental Med Sci Profess Grad Sch Oriental Med Dept Nat Prod Iksan 570749 Chonbuk South Korea

We examined the effect of sulfapyridine on mast cell-mediated immediate-type allergic reactions. Sulfapyridine (1 and 10 mu g/kg) significantly inhibited systemic allergic reaction induced by compound 48/80 in rats. Sulfapyridine (1 and 10 mu g/kg) also inhibited significantly local mast cell-mediated immediate-type allergic reactions activated by anti-dinitrophenyl (Dnp) IgE. Moreover, sulfapyridine inhibited histamine release dose-dependently in the rat peritoneal mast cells (RpMC) activated by compound 48/80 or anti-Dnp IgE. When sulfapyridine was added, the level of cAMp in RpMC, transiently and significantly increased about 4-fold compared with that of basal cells. These results indicate that sulfapyridine inhibits mast cell-mediated immediate-type allergic reactions in vivo and in vitro.

关键词：环AMp/代谢二硝基酚类/免疫学组胺释放/药物作用超敏反应速发型/预防和控制免疫细胞/药物作用肥大细胞/药物作用肥大细胞/免疫学肥大细胞/代谢被动皮肤过敏反应/药物作用腹膜腔/细胞学大鼠 Wistar 血清白蛋白/免疫学磺胺吡啶/药理学 p-甲氧基-n-苯丙胺/药理学动物大鼠

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InTRACELLULAR-DISTRIBUTIOn OF AMInES TAKEn Up BY RAT MAST-CELLS InVITRO

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ACTA pHYSIOLOGICA SCAnDInAVICA 1975年第1期95卷 133-141页

作者： BERGEnDORFF, A KAROLINSKA INST DEPT PHARMACOLSTOCKHOLMSWEDEN

Mast cells isolated from rat peritoneal and pleural cavities were incubated in vitro with radioactively-labelled histamine (Hi), 5-hydroxytryptamine (5-HT), dopamine (DA), noradrenaline (nA), tyramine (TA), phenylethylamine (phEA), trptamine (TrpA), ephedrine (Eph) or amphetamine (Amph). All these amines were taken up by the mast cells. The dose-response curves for the compound 48/80-induced release of endogenous Hi and for the various amines taken up by the cells were compared. The release curves for Hi, 5-HT, DA, nA and TA were found to be similar to that for endogenous Hi, while those for phEA, TrpA, Eph and Amph were different from that for endogenous Hi. The uptake of Hi, 5-HT, DA, phEA, TrpA and Eph into granules in mast cells was studied. Membrane-bound granules were obtained by sonication of mast cells incubated with the respective amine, followed by differential contrifugation. The amine content of these granules was then measured. Hi, 5-HT and DA were found to be mainly localized to the granules, while a smaller proportion of the phEA, TrpA and Eph was found there, the rest being located extragranularly. The present results suggest that, when taken up by rat mast cells, even amines which are not endogenous to the cells are stored in the same way as the endogenous amines Hi and 5-HT.

关键词：胺类/代谢苯丙胺/代谢多巴胺/代谢麻黄碱/代谢组胺/代谢肥大细胞/代谢去甲肾上腺素/代谢苯乙胺类/代谢血清素/代谢色胺类/代谢酪胺/代谢 p-甲氧基-n-苯丙胺/药理学动物大鼠

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ACTIOnS AnD CROSS-REACTIVITY OF AnTIALLERGIC AGEnTS AnD A CALCIUM-CHAnnEL AnTAGOnIST On RAT pERITOnEAL MAST-CELLS - DIFFEREnCE In THE ACTIOn MECHAnISMS AnD CROSS-REACTIVITY AMOnG THE AGEnTS

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AGEnTS AnD ACTIOnS 1992年第1-2期37卷 8-15页

作者： TAnIZAKI, Y OHTAnI, J KIMURA, I OKAYAMA UNIV SCH MEDDEPT MED 2OKAYAMA 700JAPAN

The actions of the antiallergic agents, disodium chromoglycate (DSCG), tranilast and ketotifen, and of a calcium channel antagonist, nicardipine, and cross-reactivity among the agents were examined by observing the inhibition of Ca-45 uptake and histamine release in rat mast cells stimulated by antigen and compound 48/80 (comp. 48/80). 1) All agents inhibited Ca-45 uptake and histamine release in mast cells stimulated by antigen. The inhibition of Ca-45 uptake by the antiallergic agents paralleled the inhibition of histamine release, while nicardipine inhibition of Ca-45 uptake was stronger than its inhibition of histamine release. 2) The action of DSCG on Ca-45 uptake and histamine release was significantly decreased in cells stimulated with antigen and phosphatidylserine (pS), while tranilast inhibition of histamine release was not affected by the addition of pS despite a significant decrease in the inhibition of Ca-45 uptake. 3) The inhibitory effect of DSCG and tranilast was significantly lower in mast cells stimulated by comp. 48/80 than in the cells stimulated by antigen. 4) Tachyphylaxis was observed in cells re-exposed to DSCG and tranilast following previous exposure to the agents. 5) Cross-reactivity was found between DSCG and tranilast.

关键词：钙/代谢色甘酸钠/药理学交叉反应组胺拮抗药/药理学组胺释放/药物作用酮替芬/药理学肥大细胞/药物作用肥大细胞/免疫学肥大细胞/代谢尼卡地平/药理学腹膜腔/细胞学磷脂酰丝氨酸类/药理学大鼠 Sprague-Dawley p-甲氧基-n-苯丙胺/药理学动物女(雌)性男(雄)性大鼠

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InFLUEnCE OF pROTEIn-KInASE-C, CAMp AnD pHOSpHATASE-ACTIVITY On HISTAMInE-RELEASE pRODUCED BY COMpOUnD-48/80 AnD SODIUM-FLUORIDE On RAT MAST-CELLS

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AGEnTS AnD ACTIOnS 1992年第1-2期37卷 1-7页

作者： BOTAnA, LM ALFOnSO, A BOTAnA, MA VIEYTES, MR LOUZAO, MC CABADO, AG HOSP VALDECILLA SERV ENDOCRINOLSANTANDERSPAIN UNIV SANTIAGO DE COMPOSTELA FAC VETDEPT FISIOLE-27002 LUGOSPAIN

We have studied the effect of protein kinase C and protein kinase A activation, and phosphatase inhibition on two different stimuli with distinct mechanisms of action. The first stimulus is compound 48/80, and its action is mediated probably by a Gi-protein, while the other is sodium fluoride, which unspecifically activates G-proteins. We established a comparative study because the action of compound 48/80 is calcium-independent, while fluoride is strictly calcium-dependent. The activation of protein kinase C was attained with the phorbol esther 12-O-tetradecanoylphorbol-13-acetate, protein kinase A was activated by increasing cAMp levels with forskolin or rolipram, and the phosphatase activity was inhibited with okadaic acid (OA), which inhibits phosphatases type 1 and 2A. Our results show that OA enhances the response to fluoride and compound 48/80 in the absence of calcium, and we conclude that calcium has a negative feedback role on the cell response. protein kinase A activation strongly inhibits the response to fluoride, and the results show a positive regulation of protein kinase C and a negative regulation of protein kinase A over fluoride response. As previously reported by other authors for the ionophore A23187, TpA notably potentiates the response to fluoride, which supports its possible modulatory role on extracellular calcium-dependent stimuli.

关键词：环AMp/代谢酶激活/生理学醚类环/药理学组胺释放/药物作用组胺释放/生理学肥大细胞/药物作用肥大细胞/生理学黑软海绵素A 磷酸二酯酶抑制剂/药理学磷蛋白磷酸酶类/拮抗剂和抑制剂磷蛋白磷酸酶类/代谢蛋白激酶C/代谢蛋白激酶类/代谢吡咯烷酮类/药理学大鼠 Sprague-Dawley 咯利普兰氟化钠/药理学十四酰佛波乙酯/药理学 p-甲氧基-n-苯丙胺/药理学动物大鼠

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Inhibition of mast cell-dependent anaphylaxis by succinic acid

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pHARMACOLOGY & TOXICOLOGY 1999年第4期84卷 154-158页

作者： Kim, HM Jung, HS Shin, HY Jung, KY Wonkwang Univ Coll Pharm Dept Oriental Pharm Iksan 570749 Chonbuk South Korea Wonkwang Univ Sch Med Dept Pharmacol & Toxicol Iksan 570749 Chonbuk South Korea

We have examined the effect of succinic acid on anaphylaxis. Succinic acid (100 mM) significantly inhibited systemic anaphylaxis induced by compound 48/80 in mice, and dose-dependently inhibited local anaphylaxis activated by anti-dinitrophenyl IgE. Further 10 and 100 mM significantly inhibited histamine release from rat peritoneal mast cells activated by compound 48/80 or anti-dinitrophenyl IgE. In addition succinic acid (0.1 and 1 mM) had a significant inhibitory effect on anti-dinitrophenyl IgE-induced tumour necrosis factor-alpha secretion from rat peritoneal mast cells. The level of cyclic AMp in rat peritoneal mast cells. when succinic acid (100 mM) was added, transiently and significantly increased about 4 times compared with that of basal cells. These results suggest a possible use of succinic acid in managing mast cell-dependent anaphylaxis.

关键词：过敏反应/预防和控制环AMp/分析组胺释放/药物作用免疫球蛋白E/免疫学肥大细胞/药物作用肥大细胞/生理学小鼠近交BALBC 大鼠 Wistar 琥珀酸/药理学肿瘤坏死因子α/分泌 p-甲氧基-n-苯丙胺/药理学动物小鼠大鼠

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InHIBITIOn OF COMpOUnD 48-80-InDUCED InTRADEnTAL SEnSORY nERVE ACTIVITY BY DISODIUM-CROMOGLYCATE AnD SEROTOnIn AnTAGOnISTS

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ACTA pHYSIOLOGICA SCAnDInAVICA 1978年第4期104卷 415-421页

作者： OLGART, L GAZELIUS, B KAROLINSKA INST DEPT PHARMACOL S-10401 STOCKHOLM 60 SWEDEN KAROLINSKA INST DEPT ENDODONT S-10401 STOCKHOLM 60 SWEDEN

Local application of compound 48/80 [p-methyloxyphenethylmethylamine formaldehyde product] induced long lasting nerve activity in intradental sensory nerves in the teeth of cats and dogs. This effect was inhibited by pretreatment with disodium cromoglycate (DSCG) given locally (2 .times. 10-2 M) or i.a. [intra-arterially, 20 mg/min]. DSCG did not have any effect on the nerve excitability per se, as judged from the responses to hypertonic naCl and air blasts applied to exposed dentin before and after administration of DSCG. Local treatment with LSD (0.1-1 mg/ml) and methysergide (0.05-0.5 mg/ml) selectively reduced or inhibited the compound 48/80 induced nerve activity. Systemic administration of methysergide (12.5 .mu.g/kg) prevented the excitatory effects of compound 48/80 but was without effect when administered during a state of established activity. Apparently, compound 48/80 has an indirect effect on intradental sensory nerves and indicate that vascular reactions take part in intradental sensory nerve excitation.

关键词：动作电位/药物作用色甘酸钠/药理学麦角酸二乙胺/药理学美西麦角/药理学感受器感觉/药物作用血清素拮抗药/药理学氯化钠/药理学牙/药物作用牙/神经支配 p-甲氧基-n-苯丙胺/药理学动物猫狗

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Local oedema and general excitation of cutaneous sensory receptors produced by electrical stimulation of the saphenous nerve in the rat

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pain 1976年第1期2卷 25-34页

作者： Chahl, Loris A. Ladd, R.J. Department of Physiology University of Queensland St. Lucia 4067 Australia

Recordings were made from multifibre strands of the saphenous nerve of rats anaesthetized with urethane and given Evan's blue intravenously. Stimulation of A plus C fibres of the saphenous nerve, but not A fibres alone, at 10 Hz for 5 min produced dye leakage in the skin of the hind limb. Stimulation of the nerve at A plus C fibre voltages with the stimulating electrodes placed distal to the recording electrodes produced reversible block of nerve impulses at the stimulating electrodes. However, when the stimulating electrodes were placed proximal to the recording electrodes, stimulation of the nerve at A plus C fibre voltages, but not at A fibre voltages, produced an increase in activity in nerve strands. An increase in activity was also observed in experiments where the contralateral saphenous nerve was stimulated. These effects were not abolished by pretreatment of rats with reserpine. The early phase of the local oedema response appeared to be reduced by local pretreatment with compound 48 80 but the excitatory action on nerve terminals was not. The effects of nerve stimulation were not mimicked by intravenous injection of 5HT, ATp or adenosine into the contralateral saphenous vein. It is suggested that several substances might be released on antidromic stimulation of C fibres and that the resulting general excitation of sensory nerve terminals might play a role in modifying the central nervous system response to nociceptive information. © 1976.

关键词：动作电位腺苷/药理学腺苷三磷酸/药理学水肿/病因学电刺激后肢小腿/神经支配肥大细胞/生理学神经元传入/生理学神经分泌感觉感受器细胞/生理学血清素/药理学皮肤/神经支配 p-甲氧基-n-苯丙胺/药理学动物男(雄)性大鼠

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A pHARMACOLOGICAL AnD HISTOLOGICAL InVESTIGATIOn OF THE InVOLVEMEnT OF MAST CELLS In CUTAnEOUS AXOn REFLEX VASODILATATIOn

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Quarterly Journal of Experimental physiology and Cognate Medical Sciences 1975年第2期60卷 123-130页

作者： Kiernan, J.A. Department of Anatomy Health Sciences Centre The University of Western Ontario London Ontario Canada

Arteriolar vasodilatation following injury to the pinna of the left ear was measured thermometrically in 43 rats. Quantitative histological assessments were made of the intact and degranulating dermal mast cells in the left and right ears of the animals. Vasodilatation, with associated increased proportions of degranulating mast cells, was found only in animals in which the sensory innervation of the left ear was intact, allowing the occurrence of axon reflexes. In rats depleted of mast cells by treatment for 5 days with compound 48/80, axon reflex vasodilatation was suppressed, indicating that mast cells are essential for the development of the flare in the triple response. Vasodilatation was also suppressed in animals injected with aspirin 20—30 min prior to injury. In these latter rats, there was no increase in the proportion of degranulating dermal mast cells It is suggested that aspirin protects the mast cells from the degranulating effect of axon reflexes. These results indicate that the arteriolar flare around the site of a cutaneous injury is mediated by the action of axon reflexes upon dermal mast cells. This conclusion is discussed in relation to other observations on vasodilatation in injured and diseased skin. © 1975 The physiological Society

关键词：阿司匹林/药理学轴突/生理学血管/药物作用血管/病理生理学去神经支配肥大细胞/细胞学肥大细胞/药物作用肥大细胞/生理学局部血流皮肤/血液供给皮肤/损伤皮肤温度血管舒缩系统/病理生理学 p-甲氧基-n-苯丙胺/药理学动物男(雄)性大鼠

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