Thirty-four dogs were anesthetized with sodium pentothal i.v. and maintained with i.v. alpha-chloralose. Neuro-muscular blockade was accomplished with gallamine triethiodide (Flaxedil). Respiratory PCO₂ was monitored continuously while artificial ventilation at a rate of 1 5 cpm and appropriate tidal volume was adjusted to maintain expiratory P CO₂ between 38 and 40 mm Hg. Blood gas analysis (P CO₂, PC₂ and pH) allowed maintenance of blood pH between 7.35 and 7.45 by periodic administration of i.v. sodium bicarbonate. Blood volume was maintained with Dextran 75 when necessary. Body temperature was monitored continuously with an esophageal thermister and maintained automatically with heating elements in the operating table. Arterial vascular isolation of the hindlimbs was accomplished by ligating all major branches of the aorta below the renal arteries except the external iliac arteries. The dog's own blood, taken from a cannula in the abdominal aorta just distal to the renal arteries, was perfused at constant flow into cannulae in the external iliac arteries through separate pumps. Each external iliac artery pressure was monitored separately (Fig. 1). A bilateral laminectomy allowed access to the L₅, ₆ and ₇ spinal segments for electrical stimulation of their ventral roots after section of the corresponding dorsal root. In 26 dogs monophasic square wave stimulation (3 to 10V, 3 msec, 8 to 20 Hz) of the ventral root of L₅, L₆ or L₇ induced:1) a decrease in the perfusion pressure (PP) in the ipsilateral hindlimb (-41.8 - 2.7 mm Hg; mean - SE); 2) a decrease in the PP in the contralateral hindlimb (-32.2 - 2.7); 3) a fall in the aortic pressure (-15.6 - 0.7). (Fig.. 3). Similar effects were observed on stimulation of the peripheral stump of the ventral root. The above described vascular effects of ventral root stimulation were resistant to intra-arterial injections of cholinergic and beta-adrenergic blocking agents administered directly into the hindlimb perfusion lines. The effectiveness of the blockades was tested with direct intra-arterial injections of the appropriate agonists. Antihistaminics (diphenhydramine and mepyramine) similarly administered and tested did abolish the response in doses which did not suppress vascular-reactivity to acetylcholine or isoproterenol. These experiments do not provide a clear explanation of the mechanisms responsible for the contralateral vasodilatation or the fall in aortic pressure. The presence of significant anastomotic channels connecting either the two hindlimbs and/or the hindlimbs with the rest of-the body was excluded. Contralateral vasodilatation might perhaps be explained by the presence of nerve fibres crossing the midline in. the fused impar ganglion of the dog. The drop in aortic pressure was not due to the activation of afferent fibres coursing in the ventral roots, nor to the peripheral release of a vasodilator substance since the onset of the phen-omenom was too fast to be explained on these grounds. The possibility exists that the drop in aortic pressure is due to the activation by the stimulated efferent fibres of some afferent nervous pathways carrying inhibitory impulses to the vasomotor centers. The present experiments, however, do not provide data supporting or excluding this hypothesis. The experimental results strongly suggest that the described vasodilatation may be mediated by histamine released directly or indirectly by the activation of fibres coursing into the lower-ventral roots. / Medicine, Faculty of / Cellular and Physiological Sciences, Department of / Graduate

Thesis (doctoral)--Katholieke Universiteit te Nijmegen.

The objective of these experiments was to investigate the effects of various membrane receptor-mediated and receptor-independent vasodilators on the resistance and capacitance vessels of conscious, unrestrained rats by measuring mean arterial pressure (MAP) and mean circulatory filling pressure (MCFP), an index of total body venous tone. ln the first set of experiments the dose-response effects of the directly-acting vasodilators nitroglycerin, sodium nitroprusside and hydralazine were determined in intact rats as well as in rats treated with the ganglionic blocker, hexamethonium. The effects of these drugs were compared with those of the vehicle, normal saline, in control rats. In intact rats, iv infusion of nitroglycerin did not alter MAP while iv infusions of nitroprusside and hydralazine caused dose-dependent decreases in MAP. In intact rats, nitroglycerin and sodium nitroprusside did. not affect MCFP while hydralazine increased MCFP. After treatment with hexamethonium all three drugs decreased MCFP, though the decreases in MCFP caused by hydralazine were not significantly different from the corresponding changes in saline-treated rats. Therefore, sodium nitroprusside and hydralazine but not nitroglycerin were effective arteriolar dilators in intact rats; all three drugs dilated arterioles in ganglionic-blocked rats, ln intact rats, the direct venodilator actions of nitroprusside and nitroglycerin were masked by endogenous sympathetic tone. When sympathetic nerve activity was attenuated, both drugs had venodilatory effects. Hydralazine, on the other hand, hao insignificant venodilatory effects both in the presence and absence of the sympathetic reflexes. In the second set of experiments we determined the dose-response effects of hexamethonium, phentolamine, prazosin and rauwolscine — the latter being non-selective ⍺, ⍺₁-selective, and ⍺₂-selective adrenoceptor antagonists, respectively — in intact rats. Prazosin and rauwolscine were also administered to rats with reflexly increased venous tone induced by the infusion of hydralazine. In intact rats iv infusions of prazosin, phentolamine and rauwolscine all caused dose-dependent decreases in MAP; only rauwolscine reduced MCFP to levels slightly below control. Hexamethonium caused a aecrease in MAP as well as a markea reduction in MCFP. After venous tone was raised by the infusion of hydralazine, both prazosin and rauwolscine dose-dependently decreased MCFP. Therefore, the resistance and capacitance vessels contain both ⍺₁- and ⍺₂-adrenoceptors. in the intact rat, however, the capacitance vessels are somewhat resistant to the effects of postjunctionally acting ⍺-antagonists in contrast to the effects of hexamethonium which acts at the level of the ganglion. / Medicine, Faculty of / Anesthesiology, Pharmacology and Therapeutics, Department of / Graduate

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 5 uppsatser.

Luk Wing Sze, Phoebe. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 173-190). / Abstracts in English and Chinese. / Abstract --- p.I / Acknowledgement --- p.XII / Publications Based on the Work in this Thesis --- p.XIII / Abbreviations --- p.IX / Table of Contents --- p.X / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1. --- Arthritis and Inflammation --- p.1 / Chapter 1.1.1. --- Tissue Reaction in Inflammation --- p.2 / Chapter 1.1.1.1. --- Mechanism of Vasodilatation --- p.2 / Chapter 1.1.1.2. --- Plasma Extravasation --- p.5 / Chapter 1.2. --- Neurogenic Inflammation --- p.8 / Chapter 1.2.1. --- Axon Reflex and Local Efferent Action of Primary Afferents --- p.9 / Chapter 1.2.2. --- Mediators of Neurogenic Inflammation --- p.10 / Chapter 1.2.3. --- Microvascular Effect of Substance P --- p.11 / Chapter 1.2.4. --- Microvascular Effect of Calcitonin Gene-Related Peptide --- p.13 / Chapter 1.3. --- Neurogenic Inflammation in the Joint --- p.16 / Chapter 1.3.1. --- Neuropeptides in Arthritic Knee --- p.16 / Chapter 1.3.2. --- Effects of Neuropeptides on Normal Knee --- p.16 / Chapter 1.3.3. --- Effects of Neuropeptides on Inflamed Knee --- p.17 / Chapter 1.3.4. --- Activation of Efferent Function of the Nerves --- p.19 / Chapter 1.4. --- Vanilloid Receptor --- p.20 / Chapter 1.4.1. --- The Use of Capsaicin as an Experimental Tool --- p.20 / Chapter 1.4.2. --- Identification of Vanilloid Receptor --- p.21 / Chapter 1.4.3. --- Molecular Biology of Vanilloid Receptor --- p.22 / Chapter 1.4.4. --- Electrophysiolgocial Properties of Vanilloid Receptors --- p.23 / Chapter 1.5. --- Activation of Vanilloid Receptors - A Detector of Physical Stimuli --- p.23 / Chapter 1.5.1. --- Exogenous Activators of Vanilloid Receptors --- p.25 / Chapter 1.5.2. --- Endogenous Activators of Vanilloid Receptors --- p.26 / Chapter 1.5.2.1. --- Anandamide as an Endovanilloid --- p.26 / Chapter 1.5.2.2. --- Other Possible Endovanilloid --- p.28 / Chapter 1.5.3. --- Biological Effects of Capsaicin --- p.29 / Chapter 1.5.3.1. --- Efferent Function: Neuropeptide Release --- p.29 / Chapter 1.5.3.2. --- Desensitization --- p.30 / Chapter 1.5.3.3. --- Neurotoxicity --- p.32 / Chapter 1.5.4. --- Biological Effects of Anandamide --- p.33 / Chapter 1.5.4.1. --- Vascular Effect of Anandamide --- p.33 / Chapter 1.5.4.2. --- Interaction of the Vanilloid and Cannabinoid System --- p.38 / Chapter 1.6. --- Aim of Study --- p.39 / Chapter Chapter 2 --- Methods --- p.40 / Chapter 2.1. --- Materials --- p.40 / Chapter 2.2. --- Protocols --- p.42 / Chapter 2.2.1. --- General Procedures --- p.42 / Chapter 2.2.2. --- Preparatory Procedure --- p.43 / Chapter 2.3. --- Measurement of Knee Joint Blood Flow --- p.46 / Chapter 2.3.1. --- Animal Preparation for Measuring Knee Joint Blood Flow --- p.48 / Chapter 2.3.2. --- Specific Procedures --- p.49 / Chapter 2.3.3. --- Image Analysis --- p.51 / Chapter 2.3.4. --- Data Analysis --- p.52 / Chapter 2.4. --- Quantification of Plasma Protein Extravasation --- p.52 / Chapter 2.4.1. --- Experimental Procedure --- p.53 / Chapter 2.4.2. --- Measurement of Evans Blue Content --- p.53 / Chapter 2.4.3. --- Measurement of Knee Joint Size --- p.54 / Chapter 2.5. --- Effect of Capsaicin on Acute Joint Inflammation --- p.54 / Chapter Chapter 3 --- Results --- p.55 / Chapter 3.1. --- Vanilloids on Knee Joint Blood Flow --- p.55 / Chapter 3.1.1. --- Capsaicin --- p.55 / Chapter 3.1.1.1. --- Cumulative Dosing of Capsaicin --- p.55 / Chapter 3.1.1.2. --- Time Course of Capsaicin-induced Vasodilatation --- p.55 / Chapter 3.1.1.3. --- VR1 Antagonists on Capsaicin-induced Vasodilatation --- p.57 / Chapter 3.1.1.4. --- NK1 Antagonists on Capsaicin-induced Vasodilatation --- p.58 / Chapter 3.1.1.5. --- CGRP Antagonist on Capsaicin-induced Vasodilatation --- p.60 / Chapter 3.1.1.6. --- Denervation on Capsaicin-induced Vasodilatation --- p.61 / Chapter 3.1.2. --- Anandamide --- p.62 / Chapter 3.1.2.1. --- Dose Responses of Anandamide --- p.62 / Chapter 3.1.2.2. --- Time Course of Anandamide-induced Vasodilatation --- p.63 / Chapter 3.1.2.3. --- VR1 Antagonist on Anandamide-induced Vasodilatation --- p.64 / Chapter 3.1.2.4. --- NK1 Receptor Antagonists on Anandamide-induced Vasodilatation --- p.65 / Chapter 3.1.2.5. --- CGRP Receptor Antagonist on Anandamide-induced Vasodilatation --- p.67 / Chapter 3.1.2.6. --- CB1 Receptor Antagonist on Anandamide-induced Vasodilatation --- p.67 / Chapter 3.1.2.7. --- CB2 Receptor Antagonist on Anandamide-induced Vasodilatation --- p.68 / Chapter 3.1.2.8. --- Anandamide Transporter Inhibitor on Anandamide-induced Vasodilatation --- p.69 / Chapter 3.1.2.9. --- Effects of Denervation on Anandamide-induced Vasodilatation --- p.70 / Chapter 3.2. --- Vanilloids on Plasma Extravasation --- p.71 / Chapter 3.2.1. --- Saline injection on Plasma Extravasation --- p.71 / Chapter 3.2.2. --- Capsaicin on Plasma Extravasation --- p.72 / Chapter 3.2.3. --- Capsaicin on Knee Joint Sizes --- p.73 / Chapter 3.2.4. --- Anandamide on Plasma Extravasation --- p.73 / Chapter 3.2.5. --- Anandamide on Knee Joint Size --- p.73 / Chapter 3.3. --- Effects of Vanilloid Agonists on Carrageenan-induced Acute Inflammation --- p.74 / Chapter 3.3.1. --- Capsaicin on Carrageenan-induced Plasma Extravasation --- p.74 / Chapter 3.3.2. --- Capsaicin on Carrageenan-induced Joint Swelling --- p.77 / Chapter 3.3.3. --- Anandamide on Carrageenan-induced Plasma Extravasation --- p.79 / Chapter 3.3.4. --- Anandamide on Carrageenan-induced Joint Swelling --- p.80 / Chapter Chapter 4 --- Discussion --- p.150 / Chapter 4.1. --- Capsaicin-induced Long Lasting Vasodilatation --- p.151 / Chapter 4.2. --- Capsaicin-induced Vasodilatation 一 a VR1 Mediated Effect? --- p.153 / Chapter 4.3. --- Substance P and CGRP in Capsaicin-induced Vasodilatation --- p.155 / Chapter 4.4. --- Anandamide-induced Vasodilatation --- p.157 / Chapter 4.5. --- VR1 in AEA-induced Vasodilatation --- p.159 / Chapter 4.6. --- Neuropeptides in AEA-induced Vasodilatation --- p.160 / Chapter 4.7. --- Cannabinoid Receptors in AEA-induced Vasodilatation --- p.161 / Chapter 4.8. --- Role of Anandamide Transporter in AEA-induced Vasodilatation --- p.163 / Chapter 4.9. --- A Neural Mechanism for Capsaicin- and AEA-induced Vasodilatation? --- p.164 / Chapter 4.10. --- Effects of Capsaicin and AEA on Plasma Extravasation --- p.167 / Chapter 4.11. --- Capsaicin and Anandamide in Acute Inflammation --- p.169 / Chapter 4.12. --- Conclusion --- p.170 / References --- p.173

一氧化氮（NO）和內皮源性超極化因子（EDHFs）是內皮衍生的血管舒張因子兩大類。 EETs是構成EDHFs的主要類型，這是由花生四烯酸通過細胞色素P450 （CYP）表氧化酶的催化活性得到。雖然這兩個EET和NO誘導血管舒張，從而降低血壓，許多報告表明，NO對EET引起的血管舒張起抑製作用。然而，不管它的重要性，有關一氧化氮對EETs的抑制作用的機理尚未完全了解。 / 在本研究中，我調查了一氧化氮對EET的負調控。通過膜電位和動脈張力測量，我們發現， 11,12-EET可引起內皮剝脫豬冠狀動脈平滑肌細胞膜超極化和血管舒張。該反應被S-亞硝基-N-乙酰青黴胺（SNAP）和8-Br-cGMP，一個NO的供體和cGMP的膜穿透物類似物，分別抑制。 SNAP和8-Br-cGMP對11,12-EET引起的細胞膜超極化和血管舒張的抑製作用被羥鈷胺，一氧化氮清除劑; ODQ ，鳥苷酸環化酶抑製劑;和KT5823 ，蛋白激酶G（PKG）抑製劑逆轉。 SNAP和8-Br-cGMP對EET反應的抑製作用也被過度供應外源性激酶底物， TAT-TRPC1S¹⁷²和TAT -TRPC1T³¹³廢除。羥鈷胺，ODQ， KT5823， TAT -TRPC1，和TAT -scrambled獨自使用不影響11,12-EET引起的細胞膜超極化和血管舒張作用。然而，獨自使用14，15-EEZE（EET的拮抗劑）抑制了11,12-EET的作用。 此外，磷酸化試驗表明， PKG可以直接在Ser172和Thr313位點磷酸化TRPC1 。此外，TRPV4 ， TRPC1 ，或KCa1.1被選擇性地抑制時，11,12-EET未能引起細胞膜超極化和血管舒張。免疫共沉澱研究表明， TRPV4 ， TRPC1和KCa1.1物理上彼此相關聯。 / 以上結果表明，NO-cGMP-PKG通路可通過TRPC1的磷酸化來抑制11,12- EETs在冠狀動脈血管平滑肌細胞上的作用。此外，TRPV4，TRPC1和KCa1.1參與11,12-EET誘導平滑肌超極化和血管舒張，他們可能互相關聯。從本研究的結果表明，NO和cGMP可通過PKG-介導的TRPC1的磷酸化，抑製EET誘導的平滑肌超極化和血管舒張。 / Nitric oxide (NO) and endothelium-derived hyperpolarizing factors (EDHFs) are two main classes of endothelium-derived vascular relaxant factors. EETs constitute a major type of EDHFs, which are derived from arachidonic acids via the catalytic activity of cytochrome P450 (CYP) epoxygenases. Although both EET and NO induce vascular relaxation, thus reduce blood pressure, numerous reports demonstrated that NO exerts an inhibitory action on EET-induced vascular relaxation. However, despite of its importance, the mechanisms related to the inhibitory effects of NO on EETs are incompletely understood. / In the present study, I investigated the scheme for negative regulation of NO on EET action. Through measurements of membrane potential and arterial tension, we showed that 11,12-EET could induce membrane hyperpolarization and vascular relaxation in endothelium-denuded porcine coronary arteries. The responses were suppressed by S-nitroso-N-acetylpenicillamine (SNAP) and 8-Br-cGMP, a NO donor and a membrane-permeant analogue of cGMP, respectively. The inhibitory actions of SNAP and 8-Br-cGMP on 11,12-EET-induced membrane hyperpolarization and vascular relaxation were reversed by hydroxocobalamin, a NO scavenger; ODQ, a guanylyl cyclase inhibitor; and KT5823, a protein kinase G (PKG) inhibitor. The inhibitory actions of SNAP and 8-Br-cGMP on EET responses were also abrogated by shielding TRPC1-PKG phosphorylation sites with excessive supply of exogenous PKG substrates, TAT-TRPC1S¹⁷² and TAT-TRPC1T³¹³. Hydroxocobalamin, ODQ, KT5823, TAT-TRPC1 and TAT-scrambled alone has no effect on 11,12-EET-induced membrane hyperpolarization and vascular relaxation. However, 14,15-EEZE (a selective EET antagonist) alone inhibits the action of 11,12-EET. Furthermore, phosphorylation assay was performed and it demonstrated that PKG could directly phosphorylate TRPC1 at Ser¹⁷² and Thr³¹³. In addition, 11,12-EET failed to induce membrane hyperpolarization and vascular relaxation when TRPV4, TRPC1, or KCa1.1 was selectively inhibited. Co-immunoprecipitation studies demonstrated that TRPV4, TRPC1 and KCa1.1 physically associated with each other in smooth muscle cells. / Taking together, our findings demonstrated that the NO-cGMP-PKG pathway may act through the phosphorylation of TRPC1 to inhibit the action of 11,12-EETs in coronary arterial smooth muscle cells. Furthermore, TRPV4, TRPC1 and KCa1.1 are critically involved in the 11,12-EET-induced smooth muscle hyperpolarization and relaxation and that they may physically associate with each other. The results from this study demonstrated that NO and cGMP could lead to PKG-mediated phosphorylation of TRPC1, resulting in an inhibition of EET-induced smooth muscle hyperpolarization and vascular relaxation. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Zhang, Peng. / "Ca" on title page is subscript. / Thesis (Ph.D.) Chinese University of Hong Kong, 2014. / Includes bibliographical references (leaves 115-133). / Abstracts also in Chinese.

Thesis (Ph. D.)--University of Missouri--Columbia, 2001. / "May 2001." Typescript. Vita. Includes bibliographical references (leaves 141-150). Also available on the Internet.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Linköping : Linköpings universitet, 2007. / Härtill 5 uppsatser.

Thesis (Ph. D.)--University of Missouri--Columbia, 2001. / Typescript. Vita. Includes bibliographical references (leaves 141-150). Also available on the Internet.