Este estudo investigou a ação do equilin em artérias mesentéricas de resistência de ratas espontaneamente hipertensas, bem como o mecanismo envolvido, comparando com o 17<font face=\"Symbol\">b-estradiol. O equilin promoveu vasodilatação equivalente à do 17<font face=\"Symbol\">b-estradiol, não demonstrando diferença nas respostas observadas em ratas intactas e ovariectomizadas. A resposta ao equilin não foi alterada pelo antagonista de receptores de estrógeno. De modo similar, a remoção do endotélio ou a inibição da adenilato ciclase, da PKA, da óxido nítrico sintase, da guanilato ciclase e da PKG não afetou o relaxamento ao equilin. Além disso, a incubação com diferentes bloqueadores de canais de K+ não alterou o relaxamento ao equilin. O equilin diminuiu a contração ao CaCl2 e ao BAYK 8644 (ativador de canais de Ca2+ do tipo-L), porém, não modificou a contração à cafeína (que promove liberação de Ca2+ do retículo sarcoplasmático), demonstrando que o efeito relaxante do equilin em artérias mesentéricas de ratas espontaneamente hipertensas se deve predominantemente ao bloqueio de canais de Ca2+ do tipo L. / The present study investigated the action of equilin in mesenteric resistance arteries from female hypertensive rats. Mechanisms contributing to equilin-induced effects were determined, comparing with 17<font face=\"Symbol\">b-estradiol. Equilin evoked vasodilatation equivalent to that of 17<font face=\"Symbol\">b-estradiol, with no difference between intact and ovariectomized rats. Equilin-induced response was not altered by the estrogen receptor antagonist. Similarly, endothelium removal or inhibition of adenylyl cyclase, PKA, NOS, guanylate cyclase or PKG did not affect the relaxation to equilin. Furthermore, the relaxation to this hormone was not altered after incubation with K+ channel blockers. Equilin reduced contraction induced by both CaCl2 and Bay K 8644 (an L-type Ca2+ channel activator), however, it was unable to alter caffeine-induced contraction (via Ca2+ release from the intracellular stores), demonstrating that equilin vasorelaxant effect in mesenteric arteries from female spontaneously hypertensive rats occurs predominantly due to blockade of L-type Ca2+ channels.

O estradiol secretado pelo folículo dominante (DOM) desempenha importante papel na luteólise da vaca. Em adição, o reconhecimento materno da prenhez (MRP) requer um ambiente uterino otimizado, o qual por sua vez depende da função luteínica e de adequadas concentrações de progesterona circulante. Os objetivos do presente estudo foi testar diferentes estratégias para otimizar a função luteínica e prevenir a influência de um DOM durante o período crítico (CP) para o MRP (de D13 a D20 após o estro o estro). Diferentes abordagens foram testadas. No exp.1, 23 vacas Nelore foram tratadas com o protocolo ovsynch para induzir um cio sincronizado (D0). As vacas receberam: Gc (n=7) - nada mais; ThCG (n=5) - 3000 IU de hCG no D5; TE2 (n=6) - 5mg de 17b-estradiol (E2) no D12; ThCG/E2 (n=5) - hCG/D5 + E2/D12. Ultra-sonografias e dosagens de progesterona plasmática durante o ciclo estral permitiram determinar que o E2 reprogramou o ciclo ovariano ao prevenir a presença (P&it;0,05) de um DOM durante quase todo CP (0,6&plusmn;0,7 dias entre D15 e D20), porém o E2 induziu a luteólise. As vacas que receberam a hCG desenvolveram corpo lúteo acessório e tiveram [P4] mais alta até o D13 (P&it;0,05). Portanto, a fase luteínica não foi prolongada No exp.2, os mesmos tratamentos foram impostos a 220 vacas Nelore (55 por grupo) após uma inseminação artificial em tempo fixo (TAI). As taxas de prenhez (PR) à TAI ou às inseminais de repasse durante uma estação com 64 dias de duração foram diminuídas (P&it;0,05) quando o E2 foi usado e a hCG não foi capaz de aumentar aquelas PR. No exp.3, vacas Red Angus no pós-parto tiveram seu estro sincronizado (D0) e receberam: nada mais (GCT, n=5) ou 200mg de gonadorrelina no D5 mais 3000 IU hCG no D13 (GRF, n=5); ou ablação de todos folículos &sup3;7mm através de aspiração folicular em D14, D17 e D20 (GRM, n=5). GRF teve luteólise retardada (18,2&plusmn;1,0b dias, 23,6&plusmn1,0a dias e 18,7&plusmn1,2b dias para GCT, GRF e GRM, respectivamente) e maior [P4] que os outros grupos. Folículos maiores que 7mm foram observados quando das aspirações. Foi possível concluir que: a) E2 permitiu consistentemente reprogramar o desenvolvimento folicular, porém causou luteólise e o seu uso trouxe efeitos negativos sobre as PR; b) a hCG melhorou a função luteínica mas não aumentou as PR; c) a ablação dos folículos &sup3;7mm não preveniu a presença do DOM no CP para o MRP e d) a associação GnRH/hCG otimizou a função luteínica, retardou a luteólise e prolongou a fase luteínica de modo que todo o CP esteve sob influência da progesterona / Estradiol secreted from the dominant follicle (DOM) plays a key role in triggering luteolysis in the cow. In addition, maternal recognition of pregnancy (MRP) requires an optimum uterine environment, which directly depends on luteal function and adequate levels of circulating progesterone. The aim of this study was to test different strategies to optimize luteal function and prevent the influence of a DOM throughout the critical period (CP) for MRP (from D13 to D20 after estrus). Different approaches were tested. In exp.1, 23 Nelore cows were treated with the ovsync protocol to induce a synchronized estrus (D0). Cows received: Gc (n=7) - nothing else; ThCG (n=5) - 3000 IU of hCG five days (D5) after estrus; TE2 (n=6) - 5mg of 17b-estradiol (E2) on D12; ThCG/E2 (n=5) - hCG/D5 + E2/D12. Ultrasound evaluation and plasmatic progesterone concentration ([P4]) throughout estrous cycle allowed to conclude: E2 reprogrammed ovarian cycle by preventing the presence (P&it;.05) of a DOM during almost all CP (0.6±.7 days within the D15 to D20 interval) but induced luteolysis; cows receiving hCG developed accessory corpus luteum and had higher [P4] up to D13 (P&it;.05). Therefore, luteolysis was not delayed and luteal phase was not prolonged. In exp.2, same treatments were imposed to 220 Nelore cows (55 per group) after a timed artificial insemination (TAI). Pregnancy rates (PR) at TAI or at AIs thereafter over a 64-day period were reduced (P&it;0.05) by using E2 and hCG was not capable to improve those PR. In exp.3, postpartum Red Angus cows were estrus synchronized (D0) and received: nothing else (GCT, n=5) or 200mg of gonadorrelin on D5 plus 3000 IU hCG on D13 (GRF, n=5); or ablation of all follicles &sup3;7mm through follicular aspiration on D14, D17 and D20 (GRM, n=5). GRF had delayed luteolysis (18.2&plusmn;1.0b days, 23.6&plusmn;1.0a days, 18.7&plusmn;1.2b days for GCT, GRF, GRM, respectively) and higher [P4] than other groups. Follicles larger than 7mm were observed in all occasions of aspiration. In could be concluded that: a) E2 allowed to consistently reschedule follicular development but caused luteolysis and its use was detrimental to PR; b) hCG improved luteal function but did not increase PR; c) ablation of 7mm follicles did not prevent a DOM throughout CP for MRP and d) GnRH/hCG association optimized luteal function, delayed luteolysis and prolonged luteal phase in such a way that all CP was under progesterone influence

Avaliamos a influência dos hormônios sexuais sobre a inflamação alérgica pulmonar em modelo experimental de asma crônica. Grupos: Basal, animais não manipulados; OVx (ovariectomizados) e Sham-OVx/alérgico (falsamente operados), sensibilizados 7 dias após a ovariectomia e desafiados com OVA a partir do 21º dia; OVx+P/alérgico, tratados com progesterona antes da sensibilização ou OVx+E/alérgico tratados com estradiol antes da sensibilização e dos desafios, seguindo o protocolo dos animais OVx. Nossos resultados mostraram que a ovariectomia reduz a migração celular para o pulmão no grupo OVx em relação ao grupo Sham-OVx/alérgico. O tratamento com estradiol aumentou o número total de células no grupo OVx+E/alérgico, enquanto o tratamento com progesterona no grupo OVx+P/alérgico reduziu a migração celular para o pulmão. Em relação a quantificação de muco nas vias aéreas, observamos redução de muco e colágeno no grupo OVx em relação ao grupo Sham-OVx/alérgico. Nos animais do grupo OVx+P/alérgico observou-se aumento na produção de colágeno em relação ao gupo sem tratamento. / We evaluated the influence of female sex hormones on the pulmonary alergic asthma in a experimental chronic asthma model.Groups: Basal, (non-manipulated animals), Ovx (Ovariectomized) and Sham- Ovx/ allergic ( animals subjected to similar manipulations except for the ovaries removal) sensitized 7 days after ovarietcomy and challenged with OVA after the 21st Day, OVX+P/allergic, treated with progesterone (200 µg/Kg) 24h before sensitization or OVx+E/allergic treated with estradiol (300 µg/Kg) 4h before sensitization and the realized challenges, according to the protocol of Ovx animal. Our results showed that the ovarietomy (Ovx) reduces cell migration towards the lungs in OVx groups when compared to the Sham-OVx/allergic group. The treatment with estradiol increased the total number of cells in the Ovx+E/alleric group, while the treatment with progesterone in the OVx+P/allergic group reduced cell migration towards the lung. About the quantification of mucus in the airways, we observed the reduction of mucus and collagen in the OVx group comparing to the non-treated group.

A asma traz remodelamento das vias aéreas por deposição de colágeno, hiperplasia mucoide, hipertrofia celular lisa e hipertrofia epitelial além de afetar a reatividade das vias aéreas. Camundongos C57Bl/6 ovariectomizadas (OVx) foram sensibilizadas e desafiadas com OVA por inalação 3 vezes por semana durante 3 ou 7 semanas. Passadas 120h pós desafio avaliou-se: reatividade traqueal, histologia, MMPs 2 e 9 e mecânica pulmonar. O grupo OVx alérgico teve a reatividade traqueal à MCh e a quantificação de muco, colágeno e músculo liso reduzida em comparação ao grupo controle (sham OVx). O tratamento com estrógeno aumentou MMP2 e o tratamento com progesterona aumentou MMP9. A mecânica ventilatória não diferiu entre os grupos. Nossos dados sugerem que o remodelamento brônquico pode ser modulado por HSFs. / Asthma is characterized by remodeling and increased airway responsiveness. Remodeling is recognized by sub-epithelial fibrosis, bronchial glands hypertrophy and goblet cell hyperplasia. We investigated role of HSF in airway remodeling and in tracheal responsiveness in a model of chronic lung inflammation. Mice were ovariectomized (OVx), sensitized and OVA challenged. Elapsed 120h of the last challenge, histological assessment and contractile response of trachea to methacholine (MCh) was performed. Total lung resistance (R) and elastance (E) were measured under mechanical ventilation. Control group consisted of Sham OVx allergic mice. Airway remodeling were significantly lower of OVx mice compared with Sham OVx allergic counterparts. OVx allergic mice showed lower tracheal responsiveness than Sham OVx. R and E were the same between groups.

Dong, Jinghui. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 106-130). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Sendo a constipação intestinal queixa freqüente em gestantes, procuramos verificar se o complexo hormonal da gestação, e em especial a progesterona, diminui a atividade muscular colônica. Para tanto, estudamos in vivo o registro do sinal mioelétrico, sob diferentes estados hormonais, em quatro pontos do cólon de ratas: ascendente proximal, ascendente distal, médio e cólon descendente por meio de implante de eletrodos na camada sero-muscular colônica. As ratas foram divididas em 5 grupos: controle, ooforectomizadas, ooforectomizadas tratadas com estrógeno, ooforectomizadas tratadas com progesterona e ratas prenhes. Os resultados mostraram uma maior atividade elétrica no cólon proximal em ratas prenhes e nas pré-tratadas com progesterona. Nas ratas prenhes a duração da atividade elétrica máxima foi, de modo significante, maior em todas as distâncias quando comparada com ratas controle. Os resultados sugerem que in vivo a progesterona e o complexo hormonal da prenhez aumentam a atividade mioelétrica do cólon proximal e que a prenhez aumenta a duração da atividade elétrica máxima no cólon em cada distância estudada, levando à formação de fezes mais desidratadas. Progesterona e prenhez não devem ser responsabilizadas por hipomotilidade do cólon, uma das hipóteses que poderia explicar a constipação intestinal em gestantes / The aim of this study was to find out if the hormonal complex of pregnancy, especially progesterone, could be responsible for decreasing colon myoelectric activity in female rats. We analyzed the records of colon myoelectric activity in vivo using the method of musculoserosal implantation of electrodes in four regions of the colon: proximal ascendent colon, distal ascendent colon, medial colon and descendent colon. The rats were divided in five groups: control, ovariectomized, ovariectomized and treated with estrogen, ovariectomized and treated with progesterone and pregnant rats. The results showed a greater electric activity in the proximal colon in pregnant and progesterone pretreated rats. In pregnant rats the duration of maximum electric activity was significantly greater in all distances studied. The results suggest that in vivo progesterone and the hormonal complex of pregnancy increase myoelectric activity of the proximal colon, and that pregnancy increases the duration of the maximum electric activity of the colon in every distance studied leading to more dehydrated fecal material. Progesterone and pregnancy should not be responsible for colon hypomotility, one of the hypothesis that could explain intestinal constipation in pregnant women

Lee Chung-hung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 113-133). / Abstracts in English and Chinese. / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- History of soybean --- p.1 / Chapter 1.2 --- Health benefits of soybean --- p.2 / Chapter 1.3 --- Introduction to flavonoids --- p.3 / Chapter 1.4 --- Bioavailability of flavonoids --- p.5 / Chapter 1.5 --- Chemistry of isoflavones --- p.6 / Chapter 1.6 --- Estrogenic property of isoflavones --- p.8 / Chapter 1.7 --- Nutritional significance of isoflavones and their glycosides --- p.8 / Chapter 1.7.1 --- Anticarcinogenic activity --- p.9 / Chapter 1.7.2 --- Antioxidative activity --- p.10 / Chapter 1.7.3 --- Cardioprotective activity --- p.13 / Chapter 1.7.4 --- Osteoprotective activity --- p.14 / Chapter 1.7.5 --- Neuroprotective activity --- p.15 / Chapter 1.7.6 --- Antiangiogenic activity --- p.16 / Chapter Chapter 2 --- Composition of Soybean Isoflavones / Chapter 2.1 --- Introduction --- p.17 / Chapter 2.2 --- Objective --- p.19 / Chapter 2.3 --- Materials and Methods --- p.20 / Chapter 2.3.1 --- Extraction and isolation --- p.20 / Chapter 2.3.1.1 --- Preparation of soybean butanol extract --- p.20 / Chapter 2.3.1.2 --- Preparation of isoflavones and their glycosides from soybean butanol extract --- p.20 / Chapter 2.3.2 --- HPLC analysis --- p.21 / Chapter 2.3.2.1 --- Sample preparation for the HPLC analysis --- p.21 / Chapter 2.3.2.2 --- HPLC analysis --- p.22 / Chapter 2.3.2.3 --- Quantification of the flavonoids and their glycosides --- p.24 / Chapter 2.4 --- Results --- p.25 / Chapter 2.4.1 --- Structural identification --- p.25 / Chapter 2.4.1.1 --- Compound 1 --- p.25 / Chapter 2.4.1.2 --- Compound 2 --- p.26 / Chapter 2.4.1.3 --- Compound 3 --- p.26 / Chapter 2.4.1.4 --- Compound 4 --- p.27 / Chapter 2.4.1.5 --- Compound 5 --- p.27 / Chapter 2.4.1.6 --- Compound 6 --- p.28 / Chapter 2.4.1.7 --- Compound 7 --- p.28 / Chapter 2.4.1.8 --- Compound 8 --- p.29 / Chapter 2.4.2 --- Quantification of isoflavones in traditional Chinese foods --- p.29 / Chapter 2.5 --- Discussion --- p.32 / Chapter Chapter 3 --- Hypocholesterolemic Effects of Soymilkin Hamsters / Chapter 3.1 --- Introduction --- p.35 / Chapter 3.1.1 --- Lipoproteins and their functions --- p.35 / Chapter 3.1.2 --- Risk factors of cardiovascular disease --- p.36 / Chapter 3.1.3 --- Hamster as an animal model of cholesterol metabolism --- p.38 / Chapter 3.2 --- Objective --- p.39 / Chapter 3.3 --- Materials and Methods --- p.40 / Chapter 3.3.1 --- Preparation of soymilk --- p.40 / Chapter 3.3.2 --- Animals --- p.40 / Chapter 3.3.2.1 --- Experiment one - Hypocholesterolemic effect of soymilk in hamsters --- p.40 / Chapter 3.3.2.1 --- Experiment two 一 The effect of fluid cross-over between soymilk and cow´ة s milk on serum cholesterol in hamsters --- p.41 / Chapter 3.3.3 --- Serum lipid and lipoprotein determinations --- p.42 / Chapter 3.3.4 --- Determination of cholesterol in organs --- p.42 / Chapter 3.3.5 --- Statistics --- p.43 / Chapter 3.4 --- Results --- p.46 / Chapter 3.4.1 --- Experiment one-Hypocholesterolemic effect of soymilk in hamsters --- p.46 / Chapter 3.4.1.1 --- Growth and food intake --- p.46 / Chapter 3.4.1.2 --- "Effect of SM and CM on TG, TC and HDL-C" --- p.46 / Chapter 3.4.1.3 --- Effect of SM and CM on non-HDL-C and ratio of non-HDL-C to HDL-C --- p.46 / Chapter 3.4.1.4 --- Effect of SM and CM on concentration of hepatic cholesterol --- p.47 / Chapter 3.4.1.5 --- "Effect of SM and CM on brain, heart and kidney cholesterol" --- p.47 / Chapter 3.4.2 --- Experiment two - The effect of fluid cross-over between soymilk and cow´ةs milk on serum cholesterol in hamsters --- p.52 / Chapter 3.4.2.1 --- Growth and food intake --- p.52 / Chapter 3.4.2.2 --- Effect of fluid cross-over on serum TC --- p.52 / Chapter 3.5 --- Discussion --- p.55 / Chapter Chapter 4 --- Antioxidant Activities of Soybean Isoflavones and Their Glycosides / Chapter 4.1 --- Introduction --- p.58 / Chapter 4.1.1 --- Role of low density lipoprotein oxidation in the development of atherosclerosis --- p.59 / Chapter 4.1.2 --- LDL oxidation --- p.61 / Chapter 4.1.3 --- Thiobarbituric acid reactive substances (TBARS) as an index of LDL oxidation --- p.62 / Chapter 4.1.4 --- "The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power""" --- p.65 / Chapter 4.1.5 --- "1,1-diphenyl-2-picrylhydrazyl (DPPH) as a measure of free radical scavenging capacity" --- p.65 / Chapter 4.1.6 --- Antioxidant and LDL oxidation --- p.65 / Chapter 4.2 --- Objective --- p.67 / Chapter 4.3 --- Materials and Methods --- p.68 / Chapter 4.3.1 --- Preparation of samples --- p.68 / Chapter 4.3.2 --- Isolation of LDL from human serum --- p.68 / Chapter 4.3.3 --- LDL oxidation --- p.69 / Chapter 4.3.4 --- TBARS assay --- p.69 / Chapter 4.3.5 --- FRAP assay --- p.70 / Chapter 4.3.6 --- DPPH assay --- p.71 / Chapter 4.3.7 --- Statistics --- p.72 / Chapter 4.4 --- Results --- p.73 / Chapter 4.4.1 --- Effects of seven individual soybean isoflavones and their glycosides on LDL oxidation --- p.73 / Chapter 4.4.2 --- The antioxidant power of individual soybean isoflavones and their glycosides in the FRAP assay --- p.73 / Chapter 4.4.3 --- Activity of individual soybean isoflavones and their glycosides as radical scavenging antioxidants --- p.74 / Chapter 4.5 --- Discussion --- p.78 / Chapter Chapter 5 --- Hypocholesterolemic Effects of Soybean Isoflavones in Ovariectomized Golden Syrian Hamsters / Chapter 5.1 --- Introduction --- p.83 / Chapter 5.1.1 --- Coronary heart disease in women --- p.83 / Chapter 5.1.2 --- Menopause as a risk factor in CHD --- p.84 / Chapter 5.1.3 --- Dietary soy in treating postmenopausal hypercholesterolemia --- p.85 / Chapter 5.2 --- Objective --- p.87 / Chapter 5.3 --- Materials and Methods --- p.88 / Chapter 5.3.1 --- Preparation of soymilk --- p.88 / Chapter 5.3.2 --- Preparation of soybean extract --- p.88 / Chapter 5.3.3 --- Animals --- p.89 / Chapter 5.3.4 --- Serum lipid determinations --- p.90 / Chapter 5.3.5 --- Determination of tissue cholesterol content --- p.90 / Chapter 5.3.6 --- Extraction of neutral and acidic sterols from fecal samples --- p.90 / Chapter 5.3.6.1 --- Determination of neutral sterols --- p.91 / Chapter 5.3.6.2 --- Determination of acidic sterols --- p.92 / Chapter 5.3.6.3 --- GLC analysis of neutral and acidic sterols --- p.92 / Chapter 5.3.7 --- Statistics --- p.93 / Chapter 5.4 --- Results --- p.96 / Chapter 5.4.1 --- Growth and food intake --- p.96 / Chapter 5.4.2 --- Effect of ovariectomy on serum TC --- p.96 / Chapter 5.4.3 --- "Effect of soymilk and soybean extract on serum TC,TG and HDL-C" --- p.96 / Chapter 5.4.4 --- Effect of soymilk and soybean extract on non-HDL-C and ratio of non- HDL-C to HDL-C --- p.97 / Chapter 5.4.5 --- Effect of soymilk and soybean extract on concentration of hepatic cholesterol --- p.97 / Chapter 5.4.6 --- Effect of soymilk and soybean extract on heart and kidney cholesterol --- p.97 / Chapter 5.4.7 --- Effect of soymilk and soybean extract on fecal neutral and acidic sterols --- p.103 / Chapter 5.5 --- Discussion --- p.106 / Chapter Chapter 6 --- Conclusion --- p.110 / References --- p.113

G蛋白偶聯雌激素受體（GPER，又名GPR30）乃最近於各種動物包括小鼠、大鼠、人類及斑馬魚中發現之新型跨膜雌激素受體。 GPER表達於脂肪組織及多種器官之中，其已被證明能與雌激素結合並介導各式快速反應及基因轉錄。針對GPER於成脂作用中角色之研究將達致對雌激素作用之更全面了解，且GPER亦有望成為治療肥胖症之一種新型標靶。 / 脂肪發育調控乃一複雜且精妙之排程，而雌激素已被證明能抑制脂肪形成，是故雌激素替代療法可舒減絶經後婦女之脂肪代謝問題。此項研究發現GPER於小鼠腹部脂肪組織及小鼠前脂肪細胞系3T3-L1中均有表達，且其信使RNA量於受誘導之3T3-L1成脂作用中錄得上調。 / 3T3-L1細胞分化作用會被名為G1之特異性GPER激動劑阻撓於克隆擴增階段（MCE），此即表明GPER有參與成脂調控之可能。通過油紅O染色發現，受G1處理之3T3-L1細胞於分化後所產生之油滴量實比其對照組為低，但此一效果能被特異性GPER小干擾RNA預處理抹除。另外，本研究以流式細胞儀及西方墨點法對細胞週期及細胞週期因子進行分析後，認為激活GPER能觸發對G1期細胞週期停滯之抑制。另一方面，受G1處理並分化中之3T3-L1細胞出現蛋白激酶B磷酸化效應，意味雌激素與GPER結合對成脂作用有雙向調節之可能性。 / 總而言之，本研究結果斷定GPER能介導雌激素對脂肪組織發育之影響，並為成脂作用之負調節因子，故此，一系列成果將有助肥胖症藥物之研發。 / A novel transmembrane estrogen receptor, G-protein coupled estrogen receptor (GPER, also known as GPR30), is recently identified in various animals including mouse, rat, human and zebrafish. GPER is expressed in many organs including fatty tissues, and has been demonstrated to mediate various rapid responses and transcriptional events upon estrogen binding. The study on the role of GPER in adipogenesis would lead to a more comprehensive understanding of estrogenic actions, with the view of identifying novel therapeutic targets for the treatment of obesity. / Regulation of adipose development is a complex and subtly orchestrated process. Estrogen has been shown to inhibit adipogenesis. Estrogen replacement therapy therefore affects fat metabolism in post-menopausal women. In this study, GPER is identified in mouse abdominal fatty tissues; and there is an up-regulation of GPER in the mouse preadipocyte cell line 3T3-L1 during induced adipogenesis. / Differentiation of 3T3-L1 cells is perturbed by the selective GPER agonist G1 at mitotic clonal expansion (MCE), indicating a possible involvement of GPER in the regulation of adipogenesis. By means of Oil-Red-O staining, the production of oil droplets in the G1-treated, differentiated 3T3-L1 cells is shown to be lower than the untreated control; and such effect is reversed by a specific siRNA knockdown of GPER. FACS analysis and Western blot analysis of cell cycle factors during MCE suggest that GPER activation triggers an inhibition of cell cycle arrest at the G1 stage. On the other hand, phosphorylation of Akt in G1-treated differentiating cells implies a possibility of bi-directional estrogenic regulation of adipogenesis via GPER. / To conclude, it is postulated that GPER mediates estrogenic actions in adipose tissues as a negative regulator of adipogenesis. These results provide insights into the development of therapeutic agents for the treatment of human obesity. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Yuen, Man Leuk. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 144-166). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract (English version) --- p.I / Abstract (Chinese version) --- p.III / Acknowledgement --- p.V / Table of Contents --- p.VII / List of Abbreviations --- p.XI / List of Tables --- p.XII / List of Figures --- p.XIII / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1. --- Obesity and adipose tissue --- p.1 / Chapter 1.1.1. --- Obesity --- p.1 / Chapter 1.1.2. --- Fat deposition --- p.3 / Chapter 1.1.3. --- Origin and development of white adipose tissue --- p.5 / Chapter 1.2. --- Adipogenesis --- p.7 / Chapter 1.2.1. --- Origins of white adipocytes --- p.7 / Chapter 1.2.2. --- Signals for adipogenesis --- p.10 / Chapter 1.2.3. --- Regulation of gene expression during adipogenesis --- p.12 / Chapter 1.2.4. --- Common adipose cell lines --- p.16 / Chapter 1.2.5. --- Mechanism of in vitro adipogenesis --- p.21 / Chapter 1.2.5.1. --- Growth arrest --- p.23 / Chapter 1.2.5.2. --- Mitotic clonal expansion --- p.23 / Chapter 1.2.5.3. --- Early and terminal differentiation --- p.24 / Chapter 1.3. --- Estrogen and adipogenesis --- p.28 / Chapter 1.4. --- G-protein coupled estrogen receptor-1 --- p.33 / Chapter 1.4.1. --- General introduction of GPER --- p.33 / Chapter 1.4.2. --- Ligands of GPER --- p.36 / Chapter 1.4.3. --- Cellular signaling of GPER --- p.38 / Chapter 1.4.4. --- Metabolic actions of GPER: A brief introduction --- p.43 / Chapter 1.4.5. --- Metabolic actions of GPER on obesity and glucose metabolism --- p.48 / Chapter 1.4.6. --- Study objectives --- p.53 / Chapter Chapter 2: --- Expression profiles and cellular localization of Gper/GPER in mouse adipose, 3T3-L1 preadipocytes and 3T3-L1 mature adipocytes --- p.54 / Chapter 2.1. --- Introduction --- p.54 / Chapter 2.1.1. --- Expression and functional roles of GPER in adipose. --- p.55 / Chapter 2.1.2. --- Swiss mouse preadipocytes 3T3-L1 --- p.57 / Chapter 2.1.3. --- Study objectives --- p.57 / Chapter 2.2. --- Materials and Methods --- p.59 / Chapter 2.2.1. --- Reagents --- p.59 / Chapter 2.2.2. --- Animal tissues --- p.59 / Chapter 2.2.3. --- Cell culture --- p.60 / Chapter 2.2.4. --- Reverse transcription polymerase chain reaction (RT-PCR) --- p.62 / Chapter 2.2.5. --- Quantitative real-time RT-PCR (qRT-PCR) --- p.66 / Chapter 2.2.6. --- SDS-PAGE and Western blot analysis --- p.68 / Chapter 2.2.7. --- Immunofluorescence assay --- p.69 / Chapter 2.2.8. --- Statistical analysis --- p.70 / Chapter 2.3. --- Results --- p.71 / Chapter 2.3.1. --- Expression of Gper/GPER in mouse visceral adipose tissues --- p.72 / Chapter 2.3.2. --- Expression profiles of Gper/GPER in undifferentiated 3T3-L1 preadipocytes and differentiated 3T3-L1 adipocytes --- p.73 / Chapter 2.3.3. --- Cellular localization of GPER in undifferentiated 3T3-L1 preadipocytes and differentiated 3T3-L1 adipocytes --- p.75 / Chapter 2.4. --- Discussion --- p.76 / Chapter Chapter 3: --- Rapid cellular responses induced by GPER activation in 3T3-L1 preadipocytes --- p.78 / Chapter 3.1. --- Introduction --- p.78 / Chapter 3.1.1. --- Rapid cellular response of estrogen via GPER --- p.79 / Chapter 3.1.2. --- Study objectives --- p.81 / Chapter 3.2. --- Materials and Methods --- p.82 / Chapter 3.2.1. --- Reagents --- p.82 / Chapter 3.2.2. --- Cell culture --- p.82 / Chapter 3.2.3. --- SDS-PAGE and Western blot analysis --- p.83 / Chapter 3.2.4. --- Statistical analysis --- p.84 / Chapter 3.3. --- Results --- p.86 / Chapter 3.3.1. --- Phosphorylation of p44/42 MAPK after time-dependent activation of GPER by ICI182,780 and G1 --- p.87 / Chapter 3.3.2. --- Phosphorylation of p44/42 MAPK after dose-dependent activation of GPER by a combination of chemical agents --- p.88 / Chapter 3.4. --- Discussion --- p.89 / Chapter Chapter 4: --- GPER activation on cell viability of 3T3-L1 preadipocytes --- p.90 / Chapter 4.1. --- Introduction --- p.90 / Chapter 4.1.1. --- Cell proliferation mediated by GPER --- p.90 / Chapter 4.1.2. --- Study objectives --- p.92 / Chapter 4.2. --- Materials and Methods --- p.93 / Chapter 4.2.1. --- Reagents --- p.93 / Chapter 4.2.2. --- Cell culture --- p.93 / Chapter 4.2.3. --- MTT assay for cell viability --- p.94 / Chapter 4.2.4. --- Statistical analysis --- p.95 / Chapter 4.3. --- Results --- p.96 / Chapter 4.3.1. --- Cell viability of 3T3-L1 after dose-dependent activation of GPER by 17β-estradiol, ICI182,780 and G1 --- p.97 / Chapter 4.4. --- Discussion --- p.99 / Chapter Chapter 5: --- GPER-mediated estrogenic action on lipid accumulation in the mature 3T3-L1 adipocytes --- p.101 / Chapter 5.1. --- Introduction --- p.101 / Chapter 5.1.1. --- Induction of differentiation in Swiss mouse preadipocyte 3T3-L1 --- p.101 / Chapter 5.1.2. --- Study objectives --- p.102 / Chapter 5.2. --- Materials and Methods --- p.103 / Chapter 5.2.1. --- Reagents --- p.103 / Chapter 5.2.2. --- Cell culture --- p.103 / Chapter 5.2.3. --- Oil-Red-O staining and measurement of absorbance --- p.105 / Chapter 5.2.4. --- Knockdown of Gper/GPER by siRNA --- p.107 / Chapter 5.2.5. --- Reverse transcription polymerase chain reaction (RT-PCR) --- p.110 / Chapter 5.2.6. --- SDS-PAGE and Western blot analysis --- p.110 / Chapter 5.2.7. --- Statistical analysis --- p.110 / Chapter 5.3. --- Results --- p.112 / Chapter 5.3.1. --- GPER activation on 3T3-L1 differentiation --- p.114 / Chapter 5.3.2. --- Knockdown of Gper/GPER in Swiss mouse preadipocyte 3T3-L1 --- p.114 / Chapter 5.3.3. --- Phosphorylation of p44/42 MAPK in Gper/GPER-knockdown 3T3-L1 after time-dependent activation of GPER by G1 --- p.117 / Chapter 5.3.4. --- Action of drugs on differentiation of Gper/GPER-knockdown 3T3-L1 --- p.117 / Chapter 5.4. --- Discussion --- p.118 / Chapter Chapter 6: --- Role of GPER in regulating cell cycle progression during mitotic clonal expansion (MCE) stage in adipogenesis of 3T3-L1 --- p.120 / Chapter 6.1. --- Introduction --- p.120 / Chapter 6.1.1. --- Differentiation stages of Swiss mouse preadipocyte 3T3-L1 --- p.121 / Chapter 6.1.2. --- Apoptosis and cell cycle progression --- p.122 / Chapter 6.1.3. --- Study objectives --- p.126 / Chapter 6.2. --- Materials and Methods --- p.127 / Chapter 6.2.1. --- Reagents --- p.127 / Chapter 6.2.2. --- Cell culture --- p.127 / Chapter 6.2.3. --- Oil-Red-O staining and measurement of absorbance --- p.129 / Chapter 6.2.4. --- Trypan blue exclusion assay for cell viability determination --- p.129 / Chapter 6.2.5. --- SDS-PAGE and Western blot analysis --- p.131 / Chapter 6.2.6. --- Flow cytometry for analysis of cell cycle progression --- p.132 / Chapter 6.2.7. --- Statistical analysis --- p.133 / Chapter 6.3. --- Results --- p.134 / Chapter 6.3.1. --- Temporal effect of GPER activation on differentiation progress of Swiss mouse preadipocyte 3T3-L1 --- p.137 / Chapter 6.3.2. --- Effect of GPER activation on cell viability during adipogenesis --- p.139 / Chapter 6.3.3. --- Effect of GPER activation on apoptosis during adipogenesis --- p.139 / Chapter 6.3.4. --- Effect of GPER activation on cell cycle distribution during induced adipogenesis --- p.140 / Chapter 6.3.5. --- Effect of GPER activation on expression of cell cycle markers during induced adipogenesis --- p.142 / Chapter 6.3.6. --- Activation of PI3K/Akt pathway by GPER stimulation during induced adipogenesis --- p.143 / Chapter 6.4. --- Discussion --- p.144 / Chapter Chapter 7: --- Conclusions and Future Perspectives --- p.148 / References --- p.155

Although there is strong evidence that thyroid tumors occur more frequently in females than in males, few studies have investigated the role sex hormones play in thyroid carcinogenesis, especially the role of estrogen (E2). This laboratory has previously shown that estrogen receptors (ERs) exist in thyroid papillary carcinoma cells. Continuing along this line of research, we studied the role of E2 and its receptors on the regulation of human thyroid cancer. / In conclusion, we have demonstrated (1) a novel mechanism by which E2 contributes to the proliferation and growth of thyroid cancer cells, (2) that E2 influences the expression of ERalpha and ERbeta differently, causing an imbalance between them, which may change the biological behavior of thyroid cancer cells, giving them the ability to proliferate and resist apoptosis by influencing the level of ERK1/2 activity and subsequently the ratio of anti-apoptotic Bcl-2 to pro-apoptotic Bax, and (3) that the subcellular localization of ERalpha and ERbeta may be a factor that contributes to the differing pathogeneses of papillary and anaplastic thyroid cancers. / To further clarify the mechanism by which E2 promotes cellular proliferation in thyroid cancer cells, we studied the localization of ERalpha and ERbeta in both KAT5 and anaplastic carcinoma cells (FRO) by immunofluorescence staining and by immunoblotting of the proteins in subcellular fractions. Cell proliferation and apoptosis were examined together with the expression of selected apoptotic proteins such as Bax, AIF and cytochrome c. We showed that the subcellular localization of ERalpha and ERbeta differed in papillary and anaplastic thyroid cancer. E2 administration led to an increase in the level of ERalpha in the nuclei of papillary cancer cells while the levels of ERbeta remained unchanged. However, the level of mitochondrial ERbeta surpassed that of ERalpha in anaplastic cancer cells. We also showed that E2 affected caspase-dependent and/or independent apoptosis via ERs in thyroid cancers. / We first studied the molecular pathways by which E2 promotes cellular proliferation in thyroid cancer cells using a human thyroid cancer cell line (KAT5) treated with E2, a selective E2 alpha receptor (ERalpha) agonist (PPT), a selective E2 beta receptor (ERbeta) agonist (DPN), an ERalpha antagonist (MPP), an E2 antagonist (ICI182780) and siRNA, which blocks ERalpha and ERbeta, by MTT assay, DNA fragmentation ELISA, BrdU cell proliferation assay and Western blot. We found that E2 and PPT gradually promoted cell proliferation by increasing the expression of ERalpha and by up-regulating the expression of Bcl-2 and pERK1/2. In contrast, we found that DPN had a negative effect on cell growth by enhancing the expression of ERbeta and Bax and by down-regulating pERK1/2 expression. At the same time, blocking ERalpha significantly reduced the E2-mediated Bcl-2 and pERK1/2 expression. On the other hand, blocking ERbeta markedly enhanced their expression. These results suggest that E2 regulates cellular growth of KAT5 cells by an ER-ERK1/2-MAPK pathway and also that E2 affects mitochondrial homeostasis. / Zeng, Qiang. / "September 2007." / Adviser: George Gong Chen. / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4616. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 136-154). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract in English and Chinese. / School code: 1307.

by Cheung Tak Chi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 117-123). / Abstract also in Chinese. / Acknowledgeements --- p.i / Abbreviations --- p.ii / Abstract --- p.v / List of Figures --- p.viii / List of Tables --- p.xiv / Contents --- p.xv / Contents / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Epidemiological Risk Factors --- p.1 / Chapter 1.1.1 --- Age --- p.1 / Chapter 1.1.2 --- Race --- p.2 / Chapter 1.1.3 --- Environmental or Migratory Factor --- p.2 / Chapter 1.1.4 --- Diet --- p.2 / Chapter 1.1.5 --- Genetics --- p.3 / Chapter 1.2 --- Regulation of Normal Prostate Development and Function --- p.4 / Chapter 1.3 --- Biochemistry and Development of Prostate Cancer --- p.6 / Chapter 1.3.1 --- Androgen-Dependent Prostate Cancer --- p.6 / Chapter 1.3.2 --- Androgen-Independent Prostate Cancer --- p.8 / Chapter 1.4 --- Classification of Prostate Cancer --- p.9 / Chapter 1.4.1 --- Stage A Prostate Cancer --- p.10 / Chapter 1.4.2 --- Stage B Prostate Cancer --- p.10 / Chapter 1.4.3 --- Stage C Prostate Cancer --- p.11 / Chapter 1.4.4 --- Stage D Prostate Cancer --- p.11 / Chapter 1.5 --- Methods for Early Detection of Prostate Cancer --- p.12 / Chapter 1.6 --- Clinical Treatment of Prostate Cancer --- p.12 / Chapter 1.6.1 --- Surgery --- p.12 / Chapter 1.6.2 --- Radiotherapy --- p.13 / Chapter 1.6.3 --- Chemotherapy --- p.13 / Chapter 1.6.4 --- Hormonal Therapy --- p.13 / Chapter 1.7 --- Objective --- p.14 / Chapter 1.8 --- Estrogen and Its Related Compounds --- p.16 / Chapter 1.8.1 --- 17β-Estradiol --- p.16 / Chapter 1.8.2 --- Tamoxifen --- p.18 / Chapter 1.8.3 --- Aromatase Inhibitor --- p.20 / Chapter 1.9 --- Anticancer Drugs --- p.23 / Chapter 1.9.1 --- Doxorubicin --- p.23 / Chapter 1.9.2 --- cis-Platinum --- p.24 / Chapter 1.10 --- Apoptotic Pathways --- p.25 / Chapter 1.10.1 --- BCL-2 /BAD Pathway --- p.26 / Chapter 1.10.2 --- FADD Pathway --- p.27 / Chapter 1.10.3 --- CAS Pathway --- p.27 / Chapter 2. --- Materials and Methods --- p.28 / Chapter 2.1 --- Materials --- p.28 / Chapter 2.2 --- Cell Lines --- p.32 / Chapter 2.3 --- Preparation of Drugs --- p.32 / Chapter 2.4 --- Drug Sensitivity Assay --- p.33 / Chapter 2.5 --- Cell Cycle Analysis --- p.35 / Chapter 2.6 --- DNA Fragmentation Assay --- p.36 / Chapter 2.7 --- Annexin Binding Assay --- p.37 / Chapter 2.8 --- Western Blot Analysis --- p.38 / Chapter 2.9 --- Data Analysis --- p.41 / Chapter 3. --- Results --- p.42 / Chapter 3.1 --- Response of Human Androgen-Independent Prostate Cancer Cells to Doxorubicin and cis-Platinum --- p.42 / Chapter 3.2 --- The Effect of 17p-Estradiol on the Growth and Anticancer Drug Sensitivity of Human Androgen-Independent Prostate Cancer Cells --- p.45 / Chapter 3.2.1 --- 17β-Estradiol on Cell Growth --- p.45 / Chapter 3.2.2 --- 17β-Estradiol on Anticancer Drug Sensitivity --- p.45 / Chapter 3.2.3 --- 17β-Estradiol and Doxorubicin on Cell Cycle Progression --- p.51 / Chapter 3.2.4 --- 17β-Estradiol and Doxorubicin Induced DNA Fragmentation --- p.57 / Chapter 3.2.5 --- 17β-Estradiol and Doxorubicin on Annexin Staining --- p.59 / Chapter 3.2.6 --- 17β-Estradiol and Doxorubicin on Apoptotic Protein Expression --- p.62 / Chapter 3.3 --- The Effect of Tamoxifen on the Growth and Anticancer Drug Sensitivity of Human Androgen-Independent Prostate Cancer Cells --- p.64 / Chapter 3.3.1 --- Tamoxifen on Cell Growth of Human --- p.65 / Chapter 3.3.2 --- Tamoxifen on Anticancer Drug Sensitivity --- p.65 / Chapter 3.3.3 --- Tamoxifen and Doxorubicin on Cell Cycle Progression --- p.71 / Chapter 3.3.4 --- Tamoxifen and Doxorubicin Induced DNA Fragmentation --- p.76 / Chapter 3.3.5 --- Tamoxifen and Doxorubicin on Annexin Staining --- p.78 / Chapter 3.3.6 --- Tamoxifen and Doxorubicin on Apoptotic Protein Expression --- p.79 / Chapter 3.4 --- The Effect of Aromatase Inhibtiors on the Growth and Anticancer Drug Sensitivity of Human Androgen-Independent Prostate Cancer Cells --- p.81 / Chapter 3.4.1 --- Aromatase Inhibitors on Cell Growth --- p.81 / Chapter 3.4.2 --- Aromatase Inhibitors on Anticancer Drug Sensitivity --- p.83 / Chapter 3.4.3 --- 4-AcA and Doxorubicin on Cell Cycle Progression --- p.93 / Chapter 3.4.4 --- 4-AcA and Doxorubicin Induced DNA Fragmentation --- p.99 / Chapter 3.4.5 --- 4-AcA and Doxorubicin on Annexin Staining --- p.100 / Chapter 3.4.6 --- 4-AcA and Doxorubicin on Apoptotic Protein Expression --- p.102 / Chapter 4. --- Discussion --- p.105 / Chapter 4.1 --- 17 β-Estradiol and Anticancer Drug Sensitivity --- p.106 / Chapter 4.2 --- Tamoxifen and Anticancer Drug Sensitivity --- p.109 / Chapter 4.3 --- Aromatase Inhibitors and Anticancer Drug Sensitivity --- p.112 / Chapter 4.4 --- DU145 Cells vs PC3 Cells --- p.115 / Chapter 5. --- Conclusion and Perspectives --- p.116 / Chapter 6. --- References --- p.117