CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O objetivo desse estudo foi verificar o efeito da suplementaÃÃo intragÃstrica com fibra solÃvel e prebiÃtico (frutooligossacarÃdeos - FOS), do ponto de vista inflamatÃrio e metabÃlico, no tecido cÃlico normal ou na vigÃncia da colite induzida por Ãcido trinitrobenzeno sulfÃnico (TNBS), em ratos. Foram usados 96 ratos da linhagem Wistar, machos, com peso mÃdio de 300g. Foram distribuÃdos aleatoriamente em dois grupos, com 48 animais. Um Grupo Estudo Sem Colite (GESC) submetido à aplicaÃÃo transanal de Ãgua ou outro Grupo Estudo Com Colite (GEC) induzida por TNBS, 20 mg mais etanol à 50%. Cada grupo foi redistribuÃdo em 3 subgrupos, de acordo com a substÃncia utilizada por infusÃo intragÃstrica: 1(Ãgua), 2(Fibra solÃvel) e 3(FOS). Todos os animais receberam essas soluÃÃes durante 14 dias antes da aplicaÃÃo transanal de Ãgua ou da induÃÃo da colite, atà o dia determinado para eutanÃsia (7 e 14 dias). Foram realizadas avaliaÃÃes do peso dos animais, relaÃÃo peso/tamanho do colo, escores macroscÃpicos e microscÃpicos, e dosagens de metabÃlitos no tecido cÃlico in vivo (concentraÃÃes de corpos cetÃnicos [3-hidroxibutirato, acetoacetato], piruvato, lactato, ATP e relaÃÃo [acetoacetato]/[3-hidroxibutirato] e [piruvato]/[lactato]. Foram avaliados o GESC, GEC, e os subgrupos Ãgua, do grupo sem colite (ESC1) e com colite (EC1). NÃo houve alteraÃÃes significantes nos diversos parÃmetros inflamatÃrios avaliados no GESC. No GEC, houve aumento na relaÃÃo peso do colo/comprimento do colo no subgrupo FOS, no 14 dia (&#961;<0,05). Com relaÃÃo ao peso dos animais, os do subgrupo GEC1, apresentaram reduÃÃo significante no 7 dia. Houve aumento na relaÃÃo peso/tamanho do colo, no escores macro e microscÃpico no subgrupo GEC1 no 7 e 14 dias (&#961;<0,05). Quanto à avaliaÃÃo metabÃlica, no GESC foi evidenciado aumento nas concentraÃÃes de acetoacetato no subgrupo fibra solÃvel no 14 dia (&#961;<0,05). Houve reduÃÃo nas concentraÃÃes de piruvato no subgrupo fibra solÃvel e FOS no 14 dia (&#961;<0,05). As concentraÃÃes de ATP foram reduzidas nos subgrupos fibra solÃvel e FOS, no 7 e 14 dias (&#961;<0,05). No GEC, as concentraÃÃes do piruvato apresentaram-se elevadas no subgrupo fibra solÃvel, no 14 dia (&#961;<0,05). As concentraÃÃes de ATP diminuÃram nos subgrupos fibra solÃvel e FOS (&#961;<0,05). Comparando os subgrupos GESC1 com GEC1, houve elevaÃÃo nas concentraÃÃes dos corpos cetÃnicos no GEC1 no 7 dia (&#961;<0,05). Houve reduÃÃo nos nÃveis de piruvato no GEC1 no 7 e 14 dias (&#961;<0,05). Observou-se reduÃÃo nas relaÃÃes [acetoacetato]/[3-hidroxibutirato] e [piruvato]/[lactato] no GEC1, no 7 dia (&#961;<0,05). Conclui-se, portanto, que a suplementaÃÃo intragÃstrica com fibra solÃvel ou com FOS, nÃo alteram os parÃmetros inflamatÃrios avaliados no 7 e 14 dias, nos colos normais ou na vigÃncia de colite, porÃm eleva a oferta de precursores para produÃÃo de energia, alterando as concentraÃÃes in vivo de ATP, tanto no tecido cÃlico normal, como na vigÃncia da colite. / The aim of this study was research the metabolic and inflammatory findings in normal colon and trinitrobenzene sulphonic acid-induced (TNBS) colitic rats which received soluble fiber and fructooligosaccharides-FOS. Ninety six Wistar male rats were used, average weight 300g. They were distributed in two groups, of 48 animals each. One group with animals without colitis (GESC) submitted to transanal water enema and another group with colitis (GEC) produced by TNBS, 20mg. Each group was distributed into three subgroups, according to the used substance: 1(water), 2(soluble fiber), 3(FOS). All the animals received 3 ml such substances twice daily into the stomach through an intragastric catether during 14 days before the transanal water enema and the colitis production until the euthanasia day (7th and 14th. Days). The animals were evaluated concerning to the weight, colon weight/size relationship, macroscopic and microscopic scores and the metabolites colon in vivo concentrations (ketone bodies [3-hydroxybutirate, acetoacetate], piruvate, lactate and ATP). The cell redox estate was also evaluated by the [acetoacetate]/[3-hydroxybutirate] and [piruvate]/[lactate] relationship. GESC and GEC were evaluated, equally the subgroups water from the group without colitis (ESC1) comparing to the colitis group (EC1). No significant differences were found in the various inflammatory evaluated parameters in GESC group. An increased colon weight/size relationship was found in GEC group in the subgroup FOS, on the 14th day (p<0,05). Concerning to the animals weight from the subgroup GEC1, a significant decreasing was found on the 7th day. An increased colon weight/size relationship was found in macroscopic and microscopic scores in the subgroup GEC1 on the 7th and 14th days (p<0,05). Concerning to the metabolic evaluation, an increased acetoacetate concentration was found in GESC, in the fiber subgroup on the 14th day (p<0,05). Decreased piruvate concentrations in the fiber and FOS subgroups were demonstrated on the 14th day (p<0,05). The ATP concentrations were reduced in the fiber and FOS subgroups on the 7th and 14th days (p<0,05). The piruvate concentrations were increased in GEC group, in the fiber subgroup, on the 14th day (p<0,05). The ATP concentrations decreased in the fiber and FOS subgroups (p<0,05). Comparing the subgroups GESC1 with GEC1, an increased cetonic corps concentration was found in GEC1 on the 7th and 14th days (p<0,05). There was reduction of piruvate levels in GEC1 on the 7th and 14th days (p<0,05). There was also reduction in acetoacetate/3-hidroxibutirate and piruvate/lactate relationship in GEC1, on the 7th day (p<0,05). Itâs concluded that the soluble fiber or FOS intragastric intake doesnât change the evaluated inflammatory parameters on the 7th and 14th days in normal colons or during colitis process, but it increases the precursors offering for energy production, modifying the ATP in vivo concentrations in normal colonic tissue and during colitis process.

Chau Shuk Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 103-115). / Abstracts in English and Chinese. / DECLARATION --- p.i / ACKNOWLEDGEMENT --- p.ii / ABBREVIATIONS --- p.iii / ABSTRACT IN ENGLISH --- p.v / ABSTRACT IN CHINESE --- p.viii / TABLE OF CONTENTS --- p.xi / Chapter CHAPTER I - --- INTRODUCTION / Chapter 1.1 --- Fluid Transport in Human Colon --- p.1 / Chapter 1.2 --- C1' Secretion across the Colonic Epithelium --- p.2 / Chapter 1.3 --- Properties of T84 Cells --- p.7 / Chapter 1.4 --- General Introduction on Extracellular Calcium Sensing Receptor (CaSR) --- p.8 / Chapter 1.5 --- Molecular Structure of CaSR --- p.9 / Chapter 1.6 --- CaSR-mediated Intracellular Signaling --- p.13 / Chapter 1.7 --- Agonists of CaSR --- p.16 / Chapter 1.8 --- Functions of CaSR --- p.18 / Chapter 1.8.1 --- Homeostatic Functions of CaSR --- p.18 / Chapter 1.8.2 --- Non-Homeostatic Functions of CaSR --- p.18 / Chapter 1.9 --- Molecular and Functional Study of CaSR Expressed in Colon --- p.19 / Chapter 1.10 --- Objectives of the Present Study --- p.21 / Chapter CHAPTER II - --- MATERIALS AND METHODS / Chapter 2.1 --- Solutions and Drugs --- p.22 / Chapter 2.2 --- Cell Culture --- p.23 / Chapter 2.3 --- Western Blot --- p.26 / Chapter 2.4 --- Simultaneous Measurement of Short-Circuit Current (Isc) and Intracellular Calcium Concentration ([Ca2+]i) --- p.27 / Chapter 2.4.1 --- Measurement of Isc and Transepithelial Resistance with Ussing Chamber --- p.27 / Chapter 2.4.2 --- Simultaneous Measurement of Isc and [Ca2+]i --- p.29 / Chapter 2.5 --- Measurement of Isc with Conventional Ussing Chamber --- p.32 / Chapter 2.6 --- Measurement of Intracellular cAMP Accumulation with Enzyme-Linked Immunosorbent Assay --- p.34 / Chapter 2.7 --- Data Analysis --- p.34 / Chapter CHAPTER III - --- RESULTS / Chapter 3.1 --- Expression of CaSR in T84 Cell Monolayers --- p.36 / Chapter 3.2 --- Poly L-arginine Induced an Increase in Isc --- p.38 / Chapter 3.2.1 --- Simultaneous Measurement of Isc and [Ca2+ ]i in Response to Poly L-arginine --- p.38 / Chapter 3.2.2 --- Interaction Between Poly L-arginine and Forskolin --- p.50 / Chapter 3.2.3 --- Ionic Mechanism of Isc Stimulated by Poly L-arginine --- p.58 / Chapter 3.2.3.1 --- Involvement of C1- in Isc Stimulated by Poly L-arginine --- p.58 / Chapter 3.2.3.2 --- Involvement of Basolateral K+ Channels in Isc Stimulated by Poly L-arginine --- p.61 / Chapter 3.2.4 --- Signaling Pathways Underlying the Isc Response to Poly L-arginine --- p.73 / Chapter CHAPTER IV - --- DISSCUSION / Chapter 4.1 --- Presence of CaSR in T84 Cell Monolayers --- p.78 / Chapter 4.2 --- Simultaneous Measurement of Isc and [Ca2+ ]i upon Application of Poly L-arginine --- p.82 / Chapter 4.3 --- Ionic Mechanism Underlying the Increase in Isc Stimulated by Poly L-arginine --- p.87 / Chapter 4.4 --- Signaling Pathway Underlying the Action of Poly L-arginine --- p.93 / Chapter 4.5 --- Does CaSR Mediate Poly L-arginine-evoked C1- Secretion across T84 Cell Monolayers? --- p.96 / Chapter 4.6 --- Future Study --- p.101 / REFERENCES --- p.103

Yue Gar-Lee Grace. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 113-120). / Abstracts in English and Chinese. / Abstract (English version) --- p.i / Abstract (Chinese version) --- p.iii / Acknowledgements --- p.v / Table of contents --- p.vi / List of figures --- p.x / List of tables --- p.xiii / List of abbreviations --- p.xiv / Chapter Chapter I: --- Introduction --- p.1 / Chapter 1.1. --- Transepithelial electrolyte transport in colon --- p.1 / Chapter 1.1.1. --- Intestinal fluid secretion --- p.1 / Chapter 1.1.2. --- Cellular mechanism of chloride secretion --- p.3 / Chapter 1.2. --- Biological activities of flavonoids --- p.6 / Chapter 1.2.1. --- Classification and general activities of flavonoids --- p.6 / Chapter 1.2.2. --- Bioavailability and pharmacokinetic properties of flavonoids --- p.8 / Chapter 1.3. --- "What is Scutellariae radix""?" --- p.9 / Chapter 1.3.1. --- Usage in Traditional Chinese Medicine --- p.9 / Chapter 1.3.2. --- Relationship with Coptidis rhizoma --- p.9 / Chapter 1.4. --- Effect of flavonoids on gastrointestinal activities --- p.12 / Chapter 1.4.1. --- Genistein and quercetin --- p.12 / Chapter 1.4.2. --- Baicalein --- p.12 / Chapter 1.5. --- Possible intracellular signaling pathway involved in the secretory response by Scutellariae radix (SR) in T84 cells --- p.14 / Chapter 1.5.1. --- Human colonic T84 cell --- p.14 / Chapter 1.5.2. --- Intracellular signaling pathway --- p.14 / Chapter 1.6. --- Aim of study --- p.17 / Chapter Chapter II : --- Methods and Materials --- p.18 / Chapter II.1. --- Culture technique of the T84 cells --- p.18 / Chapter II.2. --- Simultaneous measurement of short-circuit current (Isc) and intracellular calcium ([Ca2+]i) --- p.21 / Chapter II.2.1. --- Experimental setup --- p.21 / Chapter II.2.2. --- Preparation of the permeable supports --- p.23 / Chapter II.2.3. --- Cell seeding --- p.27 / Chapter II.2.4. --- Dye loading --- p.27 / Chapter II.2.5. --- Simultaneous measurement of Isc and [Ca2+]i- --- p.30 / Chapter II.3. --- Conventional short-circuit current (Isc) measurement --- p.34 / Chapter II.3.1. --- Experimental setup --- p.34 / Chapter II.3.2. --- Preparation of the permeable supports --- p.36 / Chapter II.3.3. --- Cell seeding --- p.36 / Chapter II.3.4. --- Measurement --- p.38 / Chapter II.4. --- Measurement of cAMP --- p.39 / Chapter II.5. --- Solutions and chemicals --- p.40 / Chapter II.6. --- Statistical analysis --- p.42 / Chapter Chapter III : --- Results --- p.43 / Chapter III. 1. --- Effects of baicalein and its interaction with calcium and cAMP-dependent secretagogues --- p.43 / Chapter III. 1.1. --- Effects of baicalein on baseline Isc and [Ca2+]i --- p.43 / Chapter III. 1.2. --- Ionic basis of baicalein-evoked Isc --- p.43 / Chapter III. 1.3. --- Effect of baicalein on carbachol-evoked Isc --- p.47 / Chapter III. 1.4. --- "Effect of baicalein on Isc stimulated by another calcium mobilizing agonist, histamine" --- p.58 / Chapter III. 1.5. --- Effect of carbachol on Isc response stimulated by baicalein --- p.61 / Chapter III. 1.6. --- Chronic effect of baicalein on carbachol-evoked increase in Isc --- p.63 / Chapter III.1.7. --- Interaction of baicalein with forskolin --- p.65 / Chapter III.2. --- Effects of baicalein on cAMP generation in T84 cells --- p.69 / Chapter III.2.1. --- Effects of baicalein on cAMP production --- p.69 / Chapter III.2.2 --- Effects of baicalein on forskolin-induced cAMP production --- p.70 / Chapter III.3. --- Effects of Scutellariae radix extract on ion transport activities in T84 cells --- p.73 / Chapter III.3.1. --- Effects of Scutellariae radix extract (SRE) on baseline Isc --- p.73 / Chapter III.3.2. --- Ionic basis of SRE-evoked Isc --- p.77 / Chapter III.3.3. --- Effects of adenylate cyclase inhibitor and PKA inhibitor --- p.77 / Chapter III.3.4. --- PKC modulation --- p.86 / Chapter III.3.5. --- Involvement of intracellular calcium --- p.86 / Chapter III.3.6. --- Involvement of cAMP --- p.94 / Chapter Chapter IV : --- Discussion --- p.98 / Chapter IV. 1. --- Effects of baicalein on ion transport in human colonic T84 cells --- p.98 / Chapter IV. 1.1. --- Roles of baicalein in chloride secretion in intestinal epithelial cells --- p.98 / Chapter IV. 1.2. --- Potentiation effect of baicalein on calcium-mediated chloride secretion --- p.100 / Chapter IV. 1.3. --- Potentiation effect of carbachol on baicalein-stimulated chloride secretion --- p.102 / Chapter IV. 1.4. --- Interaction between baicalein and forskolin --- p.104 / Chapter IV.2. --- Effects of Scutellariae radix extract on ion transport in human colonic T84 cells --- p.107 / Chapter IV.2.1 --- Characteristcs of Isc induced by Scutellariae radix extract --- p.107 / Chapter IV.2.2. --- Possible signaling mechanism involved in Isc induced by Scutellariae radix extract --- p.108 / Chapter IV.3. --- Comparison of the effects on ion transport in human colonic T84 cells produced by baicalein and Scutellariae radix extract --- p.110 / Chapter IV.3.1. --- Properties of baicalein- and Scutellariae radix extract- induced Isc response --- p.110 / Chapter IV.3.2. --- Summary --- p.111 / Chapter Chapter V : --- References --- p.113 / Publications --- p.120