Five traditional Chinese medicines were screened for their antiangiogenic activities through zebrafish angiogenic assay. Two of them, Tripterygium wilfordii and Rheum palmatum showed potential in the primary screening. T. wilfordii was selected in further study. / In the further investigation of antiangiogenic activity of triptolide on mammal systems, triptolide showed potent activity in human umbilical vein endothelial cells (HUVECs) assays including proliferation, migration and tube formation assay. It inhibited HUVEC proliferation with IC50 as low as 34 nM. It also showed more potency in HUVEC migration and tube formation assay at as low concentration as nanomolar level than SU5416, a putative VEGFR-2 inhibitor currently in clinic trials. RT-PCR and western blotting analysis showed that the underlying mechanism of triptolide correlated with down-regulation of VEGFR-2 and Tie2 expression and production. Tie2 inhibition appeared to be a later event as compared with VEGFR-2. Tie2 overexpression in HUVEC could attenuate the inhibitory effect of triptolide on HUVEC proliferation. Tie2 knockdown mimicked the inhibition activity of triptolide in tube formation assay. These phenomemon revealed that Tie2 signaling pathway plays a crucial role in triptolide-mediated angiogenesis inhibition. In in vivo Matrigel Plug assay, triptolide showed inhibition effect at as low as 100 nM. / T. wilfordii is an immune-suppressive, anti-inflammatory herb used in China for centuries. Through bioassay-guided purification, three antiangiogenic terpenoids were isolated from the ethyl acetate fraction, namely, celastrol, cangoronine and triptolide. Among them, triptolide manifested the most potent antiangiogenic activities against vessel formation. As low as 0.31microM, triptolide inhibited 20% of vessel formation, and the inhibition reached a plateau of 50% at 1.2 microM. Celatrol reduced vessel formation by more than 30% at 0.62microM, but killed 50% of the embryos at higher concentrations. Cangoronine was much weaker, inhibiting vessel formation by 20% at 2.5microM. These three components all showed stronger antiangiogenic activities than 2-methoxyestradiol, a putative compound currently under clinical trials as an antiangiogenic agent for cancer treatment, as the latter inhibited angiogenesis in zebrafish embryos by 34% at 10microM. The loss of vessel formation in the embryos treated with triptolide was further confirmed using endogenous alkaline phosphatase staining. Semi-quantitative RT-PCR analysis revealed that triptolide dose- and time-dependently reduced the mRNA expression of angiopoietin (angpt2) and tie2 in zebrafish, indicating the involvement of angpt2/tie2 signaling pathway in the antiangiogenic action of triptolide. / This research revealed that zebrafish model is a promising antiangiogenic model for both the screening of antiangiogenic agents from Chinese herbal medicine and the subsequent discovery for the drug targets. Triptolide, an anti-inflammatory component from T. wilfordii, is a potent angiogenic inhibitor through targeting VEGFR-2 and Tie2 pathways in mammal models whereas targeting ang2-tie2 pathway in zebrafish model. The anti-tumor action of triptolide was demonstrated to be partly through inhibition of tumor angiogenesis. Moreover, the potent antiangiogenic action exerted by triptolide at nanomolar dosage level gives proof that it is a promising lead compound for the development of antiangiogenic drug for cancer treatment. (Abstract shortened by UMI.) / He, Mingfang. / Adviser: Paul Pui-Hey Bot. / Source: Dissertation Abstracts International, Volume: 71-01, Section: B, page: 0247. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 84-106). / 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 Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

Capsaicin (1.3 mumol/kg, i.v.) and resiniferatoxin (48 nmol/kg, i.v.) failed to induce plasma extravasation in Suncus murinus (P>0.05). But SP (20 nmol/kg, i.v.) was able to induce salivation, and plasma extravasation in the bladder and the trachea significantly (P<0.05). NK1 receptor antagonists CP-99,994, R116301 (ID50 = 1.2 mumol/kg), and R115614 (ID50 = 1.8 mumol/kg) significantly reduced plasma leakage in the bladder (P<0.05), but not the trachea (P>0.05). R116301 (ID50 = 0.7 mumol/kg) and R115614 (ID50 = 1.2 mumol/kg) were able to inhibit the salivation response significantly (P<0.05). / R116301 and R115614 significantly reduced emesis induced by resiniferatoxin, motion, copper sulphate, and cisplatin (P<0.05), in the dose range between 23-70 mumol/kg, s.c. Both antagonists (100-300 nmol, i.c.v.) were also able to reduce cisplatin-induced emesis significantly (P<0.05), but only R116301 (10-300 nmol, i.c.v.) was able to significantly inhibit emesis induced by nicotine and copper sulphate (P<0.05). / The development of tachyldnin NK1 receptor antagonist aprepitant as an effective anti-emetic drug illustrates the importance of NK1 receptors in the emetic reflex. However, the exact anti-emetic mechanism of action is still unknown. The primary aim of the study was to investigate the relative contribution of centrally versus peripherally located NK1 receptors in the emetic reflex in Suncus murinus. The study also investigated the potential contribution of NK2 and NK3 receptors in emesis control. / The present studies demonstrated that R116301 and R115614 exhibited anti-emetic properties against various drugs, motion, and tachykinin receptor agonists. The studies also imply the existence of the classical SP subsite and the septide subsite of the NK1 receptors that are involved in the emetic reflex of Suncus murinus, which suggests that NK1 receptor antagonists that can block both subsites could become effective anti-emetic drugs. The present studies also demonstrated that both NK2 and NK3 receptors maybe involved in emesis control. It is possible that dual NK1/NK2 receptor antagonists or triple NK 1/NK2/NK3 receptor antagonists may have clinical potential as anti-emetic drugs besides the clinically used NK1 receptor antagonists. / The rank order of potency (based on pEC50 values) of tachykinin receptor agonists to contract Suncus murinus ileum was as follow: [Sar9Met(O2)11] substance P (SP) (8.1) > septide (7.9) (both NK1 receptor agonists) > neurokinin A (NKA) (7.7) > SP (7.6) > GR 64349 (NK2 receptor agonist) (7.0). For the NK1 receptor antagonists, the rank order of potency (based on pKB/pA2 values) to inhibit ileal contraction was: R116301 (7.8-8.2) ≈ R115614 (7.7-8.3) > CP-99,994 (6.4-7.3) against various NK1 receptor agonists. Furthermore, NK2 receptor antagonist saredutant (pA2 = 7.3) competitively antagonised GR 64349-induced ileal contraction. / When injected intracerebroventricularly, SP (100 nmol), septide, [Sar 9Met(O2)11] SP, NKA (all at 30 nmol), GR 64349 (10 and 30 nmol), and senktide (NK3 receptor agonist) (3-30 nmol) significantly induced emesis in Suncus murinus (P<0.05). They were also effective in inducing locomotor hyperactivity, ano-genital grooming, circling, face washing, hindlimb licking, scratching, and straub tail (3-30 nmol, P<0.05). R116301 and R115614 (both at 3 and 10 mumol/kg, s.c.) significantly antagonised some of the actions of the agonists including emesis, locomotor hyperactivity, ano-genital grooming, licking, scratching, and straub tail (P<0.05). Saredutant and NK3 receptor antagonist osanetant (both at 30 mumol/kg, s.c.) attenuated emesis induced by GR 64349 and senktide respectively (P<0.05). Saredutant (30 mumol/kg, s.c.) was also able to inhibit GR 64349-induced face washing and scratching, while osanetant (30 mumol/kg, s.c.) also significantly attenuated senktide-induced straub tail (P<0.05). / Cheng, Ho Man Frankie. / "September 2007." / Adviser: John A. Rudd. / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4691. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 194-223). / 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.

Lan Wei. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 260-301). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Photocopy. Ann Arbor, Mich. : UMI Dissertation Services, 2002. xx, 301 p. : ill. ; 22 cm. / Abstracts in English and Chinese.

Song Jingzheng. / "June 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 170-186). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

In the second part of the project, the immunology of PfMSP-133 was studied. During the invasion of merozoites, PfMSP--142 is processed into two fragments with molecular weight of 33kDa and 19kDa. The 19kDa fragment (PfMSP-119) originating from the carboxyl--terminal of PfMSP--142 is relatively more immuno-dominant in different malarial species such as P. falciparum, P. vivax and P. yoelii. In the past, only limited researches about PfMSP-1 33 were performed. Apart from its difficulty in expression, PfMSP-1 33 was also believed to be incapable of inducing protection. / Nevertheless, following the breakthrough of expressing recombinant PfMSP-1 33 in our laboratory, we have demonstrated in this study that recombinant MSP-133 can elicit antibodies with a titer up to a million. Also, we observed that MSP-133 can help MSP-119 to induce protective immunity and such effect is independent from the covalent linkage between these two proteins. Most importantly, our results show that recombinant PfMSP-133 can elicit the production of antibodies that can potentiate the inhibitory effect of anti-MSP-142 serum at high serum dilution. Results of this study give new insights in malarial vaccine development in terms of optimizing the use of adjuvant and immunization regimens. / The 42kDa carboxyl terminal fragment of Plasmodium falciparum Merozoite Surface Protein-1 (PfMSP--142) is one of the most promising candidate antigens in the development of malarial vaccine. In vivo experiments in the 1990's showed that Aotus monkeys immunized with PfMSP--142 were protected from malarial challenge. Later on, other experiments also demonstrated the possibility of using recombinant PfMSP-142 as candidate antigen for malarial vaccine. Previously, recombinant PfMSP-142 (Bvp42) was expressed with the baculovirus expression system and characterized in our laboratory. / The aim of the first part of this project is to improve the production of Bvp42. Experimental results have shown that the expression level of Bvp42 was increased under a BMN compatible baculovirus expression vector---pVL1393. Besides, a codon optimized MSP-142 nucleotide is constructed for the construction of a baculovirus carrying codon optimized MSP-142 gene and aimed for higher expression level. Unfortunately, no Bvp42 expression is observed in the transfection samples and the reason of this observation is unclear. Meanwhile, the purification of Bvp42 was also improved. Pretreatment of the hemolymph with Q--sepharose before affinity chromatography could enhance the purity of the final product. / Yuen, Sai-hang Don. / "July 2007." / Adviser: Walter K. K. Ho. / Source: Dissertation Abstracts International, Volume: 69-01, Section: B, page: 0220. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 183-195). / 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. / Abstracts in English and Chinese. / School code: 1307.

Lun Samantha Wei Man. / "August 2006." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 181-206). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

乳腺癌是女性最常見的腫瘤之一，多發生在女性絶經後，並具有雌激素依賴性。芳香化酶(CYP19)是雌激素生物合成過程中的關鍵酶，而芳香化酶抑製劑(AI)則被用於替代治療雌激素依賴性的乳腺癌。然而，AI在降低雌激素水平的同時能夠引起骨質酥鬆。此項研究的目的是找尋AI替代物。 / 黃酮類化合物是一種多酚化合物，廣泛分佈于植物中。我們先前的研究發現二氢黄酮陈皮素能夠抑制芳香化酶的生物活性，并且抑制芳香化酶高表達的乳腺癌生長。在本研究中，我們發現陳皮素在抑制腫瘤生長的同時能夠降低来曲唑引起的骨質流失。木犀草素是另外一種黄酮类化合物，它同樣能夠抑制芳香化酶的活性并減少骨流失。而與陳皮素不同的是，它能夠抑制芳香化酶的表達。在芳香化酶高表達的乳腺癌細胞(MCF-7 aro)中，木犀草素抑制芳香化酶活性的IC50是3 μM。在MCF-7 細胞中，5 μM的木犀草素能夠抑制CYP19 mRNA 的表達，螢光素酶報告實驗顯示木犀草素是通過作用于啟動子I.3和II來抑制CYP19的表達。蛋白印跡實驗表明木犀草素抑制CYP19表達的分子機制可能通過調節JNK信號通路進而減少AP-1的活性來實現。動物實驗結果顯示木犀草素能夠抑制MCF-7aro腫瘤的生長并改善來曲唑引起的骨流失。 / 環氧化酶(COX)是花生四烯酸轉化為前列腺素途徑中的一種關鍵酶。研究發現COX-2在乳腺癌組織中廣泛表達。本實驗研究了COX抑製劑在裸鼠動物模型中對乳腺癌腫瘤的作用機制。研究結果表明塞來昔布和阿司匹林在不影響血液中雌激素水平的情況下抑制乳腺癌腫瘤的生長。蛋白印迹實驗顯示這兩種藥物能夠降低腫瘤中COX-2，Cyclin A和Bcl-xL的表達。miR-98, miR-222和miR-145也能夠被塞來昔布和阿司匹林影響。 / 本研究表明陳皮素，木犀草素及COX抑制劑有潛力成為替代AI的化學治療藥物或共同治療藥物。 / Breast cancer is one of the most prevalent cancers affecting women. The majority of breast tumor growth occurred in the post-menopausal period are estrogen dependent. Aromatase (CYP19) catalyzes the rate-limiting step in the synthetic reaction of estrogen and aromatase inhibitors (AIs) are contemporary treatment for estrogen-positive breast cancer. However, estrogen-lowering drugs may promote osteoporosis. Our objective of this study further identified some alternatives for AIs. / Flavonoids are polyphenolic compounds that are ubiquitously distributed in plants. We have previously found that the flavanone hesperetin can inhibit the activity of aromatase and suppress aromatase-expressing breast tumor growth. In this project, we investigated the potential interaction between hesperetin and the AI letrozole in a mouse model. Our results showed that hesperetin could inhibit the tumor growth and reduce bone loss induced by letrozole. Similarly, another flavonoid luteolin also inhibited aromatase and prevented bone deterioration as observed in this project. In cells stably transfected with CYP19 (MCF-7aro), luteolin inhibited the aromatase activity with an IC50 value of 3μM. In addition, 5μM luteolin significantly reduced CYP19 mRNA expression in MCF-7 cells. Luciferase reporter assay revealed that luteolin could suppress CYP19 transcription at promoter regions I.3 and II. Western analysis illustrated that JNK signaling pathway was involved and deactivation of AP-1 could be the underlying molecular mechanism. Subsequently, we examined the effect in vivo. Our results showed that luteolin could inhibit the MCF-7aro tumor growth and improved bone loss induced by letrozole. / Cyclooxygenase (COX) is an enzyme responsible for the conversion of arachidonic acid into prostaglandins. It is over-expressed in breast cancer tissue and an increased expression of COX-2 was also observed in the xenograft model employed in this project. In the last study we evaluated the importance of COX-2 in breast tumor growth in this model. Our data showed that celecoxib and aspirin could significantly suppress the tumor growth without changing the plasma estrogen level. Western analysis illustrated that COX-2, Cyclin A, Bcl-xL and ER were reduced in celecoxib- and aspirin- treated tumor samples and miR-98, miR-222 and miR-145 were altered by celecoxib or aspirin. / After all, this project demonstrated that hesperetin, luteolin and COX-inhibitors could be potential chemopreventive or co-therapeutic agents. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Li, Fengjuan. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 131-148). / Abstract also in Chinese. / ACKNOWLEDGEMENTS --- p.I / ABSTRACT --- p.II / 摘要 --- p.IV / LIST OF ABBREVIATIONS --- p.V / TABLE OF CONTENTS --- p.VII / CHAPTER 1 --- p.1 / GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Types of Breast Cancer --- p.3 / Chapter 1.2 --- Nuclear Receptor Signaling Pathways in Breast Cancer --- p.5 / Chapter 1.3 --- Estrogen and Breast Cancer --- p.7 / Chapter 1.4 --- Estrogen and Bone Health --- p.8 / Chapter 1.5 --- Estrogen Biosynthesis and Aromatase --- p.10 / Chapter 1.6 --- Tissue Specific Promoter for Aromatase Expression --- p.13 / Chapter 1.7 --- Nuclear Receptors and Aromatase Promoter Regulation --- p.15 / Chapter 1.8 --- Signaling Pathway and Aromatase Expression --- p.17 / Chapter 1.9 --- Cell Cycle in Breast Cancer --- p.20 / Chapter 1.10 --- Cell Apoptosis --- p.23 / Chapter 1.11 --- Treatment of breast cancer --- p.25 / Chapter 1.12 --- Phytoestrogens --- p.29 / Chapter 1.13 --- Aim of My Study --- p.32 / CHAPTER 2 --- p.33 / MATERIALS AND METHODS --- p.33 / Chapter 2.1 --- Chemicals and Materials --- p.33 / Chapter 2.1.1 --- Chemicals --- p.33 / Chapter 2.1.2 --- Plasmids --- p.33 / Chapter 2.2 --- Cell Culture --- p.33 / Chapter 2.3 --- Aromatase Activity Assay --- p.34 / Chapter 2.4 --- Quantitative Real Time PCR --- p.36 / Chapter 2.4.1 --- RNA Isolation and cDNA Synthesis --- p.36 / Chapter 2.4.2 --- Quantitative Real Time PCR Assay --- p.37 / Chapter 2.4.3 --- MiRNA Quantitative Real Time PCR Assay --- p.38 / Chapter 2.5 --- Western Blot --- p.39 / Chapter 2.6 --- Measurement of Promoter Activity --- p.41 / Chapter 2.6.1 --- Plasmid Preparation --- p.41 / Chapter 2.6.2 --- Transient Transfection and Dual-Luciferase Assay --- p.42 / Chapter 2.7 --- Electrophoretic Mobility Shift Assay (EMSA) --- p.43 / Chapter 2.7.1 --- Nuclear protein extraction --- p.43 / Chapter 2.7.2 --- Electrophorectic Mobility Shift Assay --- p.44 / Chapter 2.8 --- Animal Experiment Design --- p.45 / Chapter 2.8.1 --- Animal Model for Hesperetin Study --- p.45 / Chapter 2.8.2 --- Animal Model for Luteolin Study --- p.46 / Chapter 2.8.3 --- Animal Model for Cycooxygenase Inhibitors Study --- p.48 / Chapter 2.8.4 --- Serum Estradiol Determination --- p.49 / Chapter 2.8.5 --- Analysis of serum lipoproteins --- p.49 / Chapter 2.8.6 --- Bone Image Acquisition and Region of Interest Selection --- p.50 / Chapter 2.9 --- Statistical Analysis --- p.50 / CHAPTER 3 --- p.51 / The citrus flavonone hesperetin prevents letrozole- induced bone loss in a mouse model of breast cancer --- p.51 / Chapter 3.1 --- Introduction --- p.51 / Chapter 3.2 --- Results --- p.54 / Chapter 3.2.1 --- Murine Body Weight and Liver Weight --- p.54 / Chapter 3.2.2 --- Effect of Hesperetin and Letrozole on Xenograft Growth in Ovariectomized Mice --- p.55 / Chapter 3.2.3 --- Hesperetin Reduced Plasma Estradiol Concentration --- p.58 / Chapter 3.2.4 --- PS2 mRNA Expression in Tumor --- p.59 / Chapter 3.2.5 --- Uterine Wet Weight --- p.60 / Chapter 3.2.6 --- Hesperetin Prevent Bone Deterioration Induced by Letrozole --- p.61 / Chapter 3.3 --- DISCUSSION --- p.63 / CHAPTER 4 --- p.66 / dIETARY FLAVONOID LUTEOLIN ON cyp19 transcription in the breast cancer cells mcf-7 --- p.66 / Chapter 4.1 --- Introduction --- p.66 / Chapter 4.2 --- Results --- p.68 / Chapter 4.2.1 --- Inhibitory Effect of Luteolin on Aromatase Activity --- p.68 / Chapter 4.2.2 --- Luteolin Reduced Aromatase mRNA Expression in MCF-7 Cells --- p.70 / Chapter 4.2.3 --- Effect of Luteolin on Promoter I.3/II Activity of CYP19 in MCF-7 Cells --- p.71 / Chapter 4.2.4 --- The Effect of Luteolin on Truncation CYP19 Gene Reporter Assay --- p.72 / Chapter 4.2.5 --- Luteolin Reduced AP-1 Binding in Promoter I.3/II DNA Fragment --- p.74 / Chapter 4.2.6 --- Inhibitory Effect of Luteolin on Protein Kinase Signaling --- p.76 / Chapter 4.3 --- Discussion --- p.78 / CHAPTER 5 --- p.83 / interaction OF LUTEOLIN and letrozole in a postmenopausal breast cancer model --- p.83 / Chapter 5.1 --- Introduction --- p.83 / Chapter 5.2 --- Results --- p.86 / Chapter 5.2.1 --- Luteolin and letrozole treatment had no effect on mouse body weight and liver weight --- p.86 / Chapter 5.2.2 --- Effect of luteolin and Letrozole on Xenograft Growth in Ovariectomized Mice --- p.88 / Chapter 5.2.3 --- Luteolin reduced plasma estradiol concentration --- p.91 / Chapter 5.2.4 --- Luteolin Counteracted Uterine Weight Reduction under Letrozole Treatment --- p.92 / Chapter 5.2.5 --- Luteolin Prevented Bone Deterioration Induced by Letrozole --- p.93 / Chapter 5.2.6 --- The Effect of Luteolin on Plasma TC and TG --- p.95 / Chapter 5.2.7 --- Luteolin Increased HDL Level and Reduced the Ratio of LDL/HDL --- p.97 / Chapter 5.2.8 --- Effect of Luteolin on Cell Cycle and Apoptotic Protein Expression --- p.99 / Chapter 5.3 --- DISCUSSION --- p.104 / CHAPTER 6 --- p.107 / cyclooxygenase inhibitors suppresse breast tumor growth in NUDE MICE --- p.107 / Chapter 6.1 --- Introduction --- p.107 / Chapter 6.2 --- Results --- p.109 / Chapter 6.2.1 --- Celecoxib and aspirin treatment had no effect on mouse body weight and liver weight --- p.109 / Chapter 6.2.2 --- Effect of celecoxib and aspirin on Xenograft Growth in Ovariectomized Mice --- p.111 / Chapter 6.2.3 --- Celecoxib and aspirin had no effect on plasma estradiol concentration --- p.113 / Chapter 6.2.4 --- Celecoxib and Aspirin Had no Effect on Uterine Weight --- p.114 / Chapter 6.2.5 --- Protein expression of COX-2, Cell cycle-related and cell Apoptotic Genes --- p.115 / Chapter 6.2.6 --- Detection of Related miRNA Expression Level in Tumors --- p.118 / Chapter 6.2.7 --- c-Myc mRNA Expression Level were Regulated in Tumors --- p.121 / Chapter 6.3 --- DISCUSSION --- p.124 / CHAPTER 7 --- p.127 / SUMMARY --- p.127 / REFERENCE --- p.131

背景及研究目的：據報導薑黃素（curcumin）在阿爾茨海默癥（老年癡呆癥）的動物模型中表現出有效性。這可能與其能阻抑β 澱粉樣蛋白的聚集有關，而β 澱粉樣蛋白正是老年癡呆癥中達成共識的神經毒性物質。然而，薑黃素的水溶性很差，這一弱點直接限制其在作為口服藥物的生物利用度。羥丙基-β-環糊精是一種由七個單糖分子鍵合成的環狀分子，該環具有親水性的外部和疏水性的內部，這一特點使得疏水性藥物可以裝入環糊精分子內部從而提高該藥物的親水性，進而提高藥物的口服吸收率。 / 方法：包合物在50 攝氏度下，由摩爾比為1:2 的薑黃素和羥丙基-β-環糊精經過6 小時的攪拌而形成。其后对包合物的物化性質與相同摩爾比的薑黃素與羥丙基-β-環糊精的混合物的物化特性进行了对比以证实包合物的形成，诸如水溶性，掃描電鏡下的形態以及傅利葉紅外圖譜特性。接著用經包合物和普通薑黃素粉末喂食的SD 大鼠做藥代動力學研究，取出大鼠血樣然之後用高效液相色譜-質譜聯用法對薑黃素進行定量，計算出達峰時間（T{U+2098}{U+2090}{U+2093} ）， 峰濃度（C{U+2098}{U+2090}{U+2093} ）以及 0 到4 小時藥時曲綫下面積 （AUC₀→₄{U+2095} ）從而比較包合物較普通薑黃素是否有優越之處。之後，用Tg2576 轉基因小鼠做短期研究，此种小鼠能過度表達β 澱粉樣蛋白，是一種被廣泛應用于老年癡呆癥相關科研的動物模型，經過連續7 日的喂食之後，對小鼠的β 樣澱粉蛋白進行觀察。進一步，將用能同時過度表達β 樣澱粉蛋白與神經纖維結（NTFs）的Tau/APP 轉基因小鼠做長達兩月的藥效學實驗，在實驗始末配以关联恐惧调件反应测试（CFC）和八臂迷宮（Radial Arm Maze）來對實驗小鼠的記憶失進行定量分析。處死小鼠之後，對β 樣澱粉蛋白與神經纖維結（NTFs）進行定量分析從而確定包合物在藥效學上的優勢。 / 結果：在包合的過程中，大部份的薑黃素被整合到包合物之中。通過傅利葉紅外圖譜和掃描電鏡照片都可以觀察到包合物與混合物的顯著不同。在藥代動力學研究中，普通薑黃素粉末的達峰時間為22 分鐘左右，而包合物是40 分鐘，同時，經過包合，峰濃度也提高了3 倍左右，藥時曲綫下面積提高了2 倍以上。在用Tg2576 進行的為期一周的短期實驗中，觀察不到包合物和普通薑黃素在β 樣澱粉蛋白清除方面有明顯差別，然後通過體內染色卻可以看到經包合物喂食的小鼠腦切片中可以觀察到更多的來自薑黃素的螢光信號。在2 個月的長期實驗中，就关联恐惧调件反应测试和八臂迷宮實驗的結果來看，可以觀察到包合物有更好延遲TAPP 小鼠記憶失過程的趨勢但無顯著性，除此之外，對處死后的小鼠腦部進行分析，其β 澱粉樣蛋白與神經纖維節的含量分析結果也和行為測試具有一致性。 / 結論：用羥丙基-β-環糊精對薑黃素進行包合確實可以通過增加薑黃素的水溶性從而提高其生物利用度，讓更多的薑黃素通過血腦屏障進入大腦進而與β 澱粉樣蛋白進行結合。然後短期實驗無法表明包合物具有β 澱粉樣蛋白清除效應。而長期實驗中行為實驗和處死後大腦分析顯示出較普通薑黃素而言包合物具有有限的優點，如果要證明這一優點確實存在，可能需要更長時間的喂食與另外劑量。 / Background: Curcumin is reported to be an effective treatment in animal models of Alzheimer’s disease (AD), possibly by inhibiting aggregation of amyloid-β peptides, which can be neurotoxic.However, curcumin is poorly soluble in water, limiting its oral bioavailability. Hydroxypropyl-β-cyclodextrin (HP-β-CD), a cyclic oligosaccharide made of seven sugar molecules bound togetherin a ring has a hydrophobic exterior and a hydrophobic interior, within which curcumin can reside, thus increasing the aqueous solubility of curcumin. This study aims to solve the problem of poor water-solubility of curcumin using HP-β-CD. / Method: The inclusion complexes were formed by stirring a suspension of curcumin and HP-β-CD at a molar ratio of 1:2 at 50°C for 6 hr. Physicochemical properties, including watersolubility,morphology under scanning electron microscopy (SEM) and the Fourier Transform Infrared (FTIR) spectrum, varied between the inclusion complex and a physical mixture of the two compounds. The inclusion complex and curcumin powder were fed to Sprague Dawley rats for pharmacokinetic studies, from which blood samples were analyzed using LC/MS/MS, and relevant parameters such as T{U+2098}{U+2090}{U+2093}, C{U+2098}{U+2090}{U+2093} and AUC₀→₄{U+2095} were calculated to study the effect of HP-β-CD on the bioavailability of curcumin. To evaluate the pharmacodynamic effects, Tg2576 mice, which over express amyloid-β, were treated for 7 consecutive days with curcumin powder or inclusion complexes. Further, to examine effects of long-term treatment, Tau/APP mice, a commonly used AD model producing both amyloid-β and mutant tau proteins, were treated for 2 months. Behavior tests were conducted at the beginning and end of the long-term treatment to quantify the memory loss of the mice, and post mortem analyses, including quantification of amyloid-β plaques and neurofibrillary tangles, were performed after sacrificing the mice. / Result: The majority of curcumin was integrated into complexes. FTIR profiles and SEM photographs of complexes displayed significant differences from the physical mixture. In the pharmacokinetic study, the concentration of curcumin in the control group peaked at around 22.5 min, while that of inclusion complexes peaked around 40 min. The maximum concentration of curcumin trebled and the area under the curve from 0 to 4 hours more than doubled. For shortterm treatment in Tg2576 mice, paraffin sections stained with Thioflavin T, a dye detecting amyloid-β plaques, showed no obvious difference between mice treated with curcumin powder or complexes; however brain sections from complex-treated mice had more fluorescence signal from curcumin than did mice treated with curcumin powder. For long-term treatment, in terms of the results of contextual fear conditioning and radial arm maze, there was a trend toward inclusion complexes delaying the memory loss of Tau/APP mice more effectively than curcumin powder. Compared to curcumin powder, complexes tended to reduce the number of plaques and neurofibrillary tangles. / Conclusion: Complexation with HP-β-CD can significantly enhance curcumin bioavailability by increasing its water-solubility, allowing more curcumin to penetrate the blood brain barrier to bind to amyloid-β plaques. However, short-term treatment showed no advantage of inclusion complexes in clearing amyloid-β plaques. The results of behavior tests and post-mortem studies from the 2-month long-term treatment indicated limited superiority of inclusion complexes over curcumin powder. A longer feeding period or altered dosage or both might be necessary to enhance the effect of curcumin inclusion complexes. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Liu, Hao. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 104-117). / Abstracts also in Chinese. / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Overview of Alzheimer’s disease --- p.1 / Chapter 1.1.1 --- The pathological mechanisms of Alzheimer’s disease --- p.2 / Chapter 1.1.2 --- Anti-Alzheimer’s disease drugs --- p.3 / Chapter 1.2 --- Curcumin as a potential anti-AD agent --- p.5 / Chapter 1.2.1 --- Actions of curcumin on Aβ --- p.6 / Chapter 1.2.2 --- Anti-inflammatory and anti-oxidant activities of curcumin in AD --- p.8 / Chapter 1.2.3 --- Activities of curcumin to combat metal toxicity --- p.9 / Chapter 1.3 --- The physicochemical properties of curcumin and limitations of curcumin in clinical usage --- p.12 / Chapter 1.4 --- Cyclodextrin in pharmaceutical usage --- p.18 / Chapter 1.5 --- Aims of the study --- p.23 / Chapter Chapter 2 --- Preparation and characterization of curcumin-HP-β-CD complexes --- p.24 / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Phase solubility study --- p.24 / Chapter 2.1.2 --- Preparation of solid complexes --- p.28 / Chapter 2.1.3 --- Characterization of solid complexes --- p.29 / Chapter 2.1.4 --- Previous work on the curcumin HP-β-CD inclusion complex --- p.31 / Chapter 2.2 --- Methodology --- p.33 / Chapter 2.2.1 --- Phase solubility test --- p.33 / Chapter 2.2.2 --- Preparation of curcumin-CD complexes --- p.33 / Chapter 2.2.3 --- Recovery --- p.35 / Chapter 2.2.4 --- Differential solubility --- p.36 / Chapter 2.2.5 --- SEM Studies --- p.37 / Chapter 2.2.6 --- Infrared --- p.37 / Chapter 2.3 --- Results and discussion --- p.38 / Chapter 2.3.1 --- Phase solubility analysis --- p.38 / Chapter 2.3.2 --- Recovery test --- p.40 / Chapter 2.3.3 --- Differential solubility --- p.42 / Chapter 2.3.4 --- SEM study --- p.44 / Chapter 2.3.5 --- Infrared study --- p.45 / Chapter Chapter 3 --- Staining of amyloid plaques in Tg2576 mice after oral administration of curcumin-HP-β-cyclodextrin inclusion complex --- p.47 / Chapter 3.1 --- Introduction --- p.47 / Chapter 3.2 --- In vitro staining of amyloid plaques by thioflavin T and curcumin --- p.49 / Chapter 3.2.1 --- Thioflavin T and curcumin specifically binding to amyloid plaques --- p.49 / Chapter 3.2.2 --- Chemicals --- p.51 / Chapter 3.2.3 --- Method --- p.51 / Chapter 3.3 --- In vivo staining of amyloid plaques after oral administration of curcumin-HP-β-CD inclusion complex --- p.53 / Chapter 3.3.1 --- Animal treatment --- p.53 / Chapter 3.3.2 --- Method --- p.54 / Chapter 3.4 --- Results and discussion --- p.56 / Chapter 3.4.1 --- In vitro staining of amyloid plaques by thioflavin T and curcumin --- p.56 / Chapter 3.4.2 --- In vivo staining of amyloid plaques by curcumin --- p.57 / Chapter 3.4.3 --- Plaque removal effects after short-term feeding --- p.58 / Chapter Chapter 4 --- Pharmacokinetic study of curcumin in Sprague-Dawley (SD) rats after oral administration of curcumin-HP-β-cyclodextrin inclusion complex --- p.61 / Chapter 4.1 --- Introduction --- p.61 / Chapter 4.1.1 --- Previous pharmacokinetic study of curcumin-HP-β-CD inclusion complex --- p.61 / Chapter 4.1.2 --- Previously established LC/MS/MS methods for quantification of curcumin in SD rat plasma sample --- p.62 / Chapter 4.2 --- Development and validation of LC/MS/MS assay --- p.63 / Chapter 4.2.1 --- Chemicals --- p.63 / Chapter 4.2.2 --- Instrumentation and chromatographic conditions --- p.63 / Chapter 4.2.3 --- Preparation of standard solutions --- p.63 / Chapter 4.2.5 --- Validation of the assay method --- p.64 / Chapter 4.3 --- Pharmacokinetic profile of curcumin in SD rats --- p.65 / Chapter 4.3.1 --- Animals --- p.65 / Chapter 4.3.2 --- Animal treatment and blood sampling --- p.65 / Chapter 4.3.3 --- Plasma sample analysis --- p.65 / Chapter 4.3.4 --- Data analysis --- p.66 / Chapter 4.4 --- Result and discussion --- p.67 / Chapter 4.4.1 --- Mass spectrum --- p.67 / Chapter 4.4.2 --- Chromatography and specificity --- p.69 / Chapter 4.4.3 --- Linearity and sensitivity --- p.71 / Chapter 4.4.4 --- Method validation---Precision and accuracy --- p.71 / Chapter 4.4.5 --- Method validation---Liquid-liquid extraction recovery --- p.72 / Chapter 4.4.6 --- Pharmacokinetics parameters --- p.73 / Chapter Chapter 5 --- Long-term effects of curcumin-HP-β-CD inclusion complex on Alzheimer’s disease model mice --- p.79 / Chapter 5.1 --- Introduction --- p.79 / Chapter 5.1.1 --- Tau/APP trangenic mouse --- p.79 / Chapter 5.1.2 --- Memory --- p.80 / Chapter 5.1.3 --- The role of the hippocampus in memory --- p.81 / Chapter 5.1.4 --- Memory task contextual fear conditioning (CFC) --- p.82 / Chapter 5.1.5 --- Memory task radial arm maze (RAM) --- p.83 / Chapter 5.2 --- Methods --- p.85 / Chapter 5.2.1 --- Animal treatment --- p.85 / Chapter 5.2.2 --- Contextual fear conditioning test --- p.85 / Chapter 5.2.3 --- Radial arm maze test --- p.87 / Chapter 5.2.4 --- Post-mortem analysis amyloid plaque removal effects of the curcumin-HP-β-CD inclusion complex --- p.88 / Chapter 5.2.5 --- Post-mortem analysis neurofibrillary tangle removal effects of the curcumin-HP-β-CD inclusion complex --- p.90 / Chapter 5.3 --- Results and discussion --- p.92 / Chapter 5.3.1 --- Mortality of the mice --- p.92 / Chapter 5.3.2 --- Contextual fear conditioning test --- p.93 / Chapter 5.3.3 --- Radial arm maze (RAM) test --- p.95 / Chapter 5.3.4 --- Post-mortem analysis amyloid plaque removal effects of the curcumin-HP-β-CD inclusion complex --- p.97 / Chapter 5.3.5 --- Post-mortem analysis neurofibrillary tangle removal effects of the curcumin-HP-β-CD inclusion complex --- p.99 / Chapter Chapter 6 --- Conclusions and future perspective --- p.100 / Chapter 6.1 --- Conclusions --- p.101 / Chapter 6.2 --- Future perspective --- p.103 / References --- p.104

Zhang Jie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 54-74). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.ii / ABSTRACT (Chinese) --- p.iv / ABBREVIATION --- p.vi / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Overview of the study --- p.1 / Chapter 1.2 --- Secretin --- p.3 / Chapter 1.2.1 --- Discovery / Chapter 1.2.2 --- Molecular biology / Chapter 1.2.3 --- Biosynthesis and localization / Chapter 1.2.4 --- Function / Chapter 1.3 --- Secretin receptor --- p.8 / Chapter 1.3.1 --- Molecular biology / Chapter 1.3.2 --- Localization / Chapter 1.3.3 --- Signal transduction pathway / Chapter 1.4 --- Secretin and autism --- p.13 / Chapter 1.5 --- AMPA receptor --- p.15 / Chapter 1.5.1 --- Molecular biology / Chapter 1.5.2 --- Localization / Chapter 1.5.3 --- Pharmacological property / Chapter 1.5.4 --- Function / Chapter 1.6 --- Cerebellum --- p.20 / Chapter 1.6.1 --- Structure of the cerebellar cortex / Chapter 1.6.2 --- Neurons of the cerebellar cortex / Chapter 1.6.2.1 --- Granule cells / Chapter 1.6.2.2 --- Purkinje cells / Chapter 1.6.2.3 --- Basket and stellate cells / Chapter 1.6.2.4 --- Golgi cells / Chapter 1.6.3 --- Intrinsic circuitry of the cerebellar cortex / Chapter CHAPTER 2 --- METHODS AND MATERIALS --- p.25 / Chapter 2.1 --- Brain slice preparation and maintenance --- p.25 / Chapter 2.2 --- Experimental set-up --- p.26 / Chapter 2.2.1 --- Visualization of neurons / Chapter 2.2.2 --- Electrophysiological recordings / Chapter 2.2.3 --- Evoked stimulation / Chapter 2.2.4 --- Drug preparation and administration / Chapter 2.3 --- Data analysis --- p.29 / Chapter 2.3.1 --- Construction of dose-response curve / Chapter 2.3.2 --- Analysis of synaptic currents / Chapter 2.3.3 --- Statistics / Chapter CHAPTER 3 --- RESULTS --- p.31 / Chapter 3.1 --- Basic characteristics of IPSCs recorded from PCs --- p.31 / Chapter 3.1.1 --- Spontaneous IPSCs / Chapter 3.1.2 --- Miniature IPSCs / Chapter 3.1.3 --- Evoked IPSCs / Chapter 3.1.4 --- Rundown of IPSCs / Chapter 3.2 --- Electrophysiological effects of secretin --- p.33 / Chapter 3.2.1 --- Effects of secretin on evoked IPSCs and EPSCs / Chapter 3.2.2 --- Effects of secretin on spontaneous IPSCs / Chapter 3.2.3 --- Effects of secretin on miniature IPSCs / Chapter 3.3 --- Mechanisms of secretin as a neuropeptide --- p.37 / Chapter 3.3.1 --- Non-involvement of a postsynaptic site of action / Chapter 3.3.2 --- Non-involvement of calcium influx / Chapter 3.3.3 --- Involvement of cAMP second messenger / Chapter 3.3.4 --- Involvement of presynaptic AMP A receptors / Chapter 3.3.4.1 --- Glutamate-mediated action of secretin / Chapter 3.3.4.2 --- Effects of AMPA on miniature IPSCs / Chapter 3.3.4.3 --- Pharmacological evidence / Chapter CHAPTER 4 --- DISCUSSION --- p.45 / Chapter 4.1 --- Secretin as a novel neuropeptide --- p.45 / Chapter 4.2 --- Mechanisms of secretin --- p.46 / Chapter 4.3 --- Physiological role of secretin in the cerebellum --- p.52 / Chapter 4.4 --- Secretin and autism --- p.52 / REFERENCES --- p.54

Chan Hoi Yun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 131-144). / Abstracts in English and Chinese. / DECLARATION --- p.i / ACKNOWLEDGMENTS --- p.ii / ABBREVIATIONS --- p.iii / ABSTRACT IN ENGLISH --- p.v / ABSTRACT IN CHINESE --- p.viii / CONTENTS --- p.xi / Chapter Chapter 1 --- Introduction / Chapter 1.1. --- Steroid Hormones --- p.1 / Chapter 1.1.1. --- Synthesis of estrogens and progesterone --- p.1 / Chapter 1.2. --- Cellular Mechanisms of Female Steroid Hormones --- p.5 / Chapter 1.2.1. --- Genomic actions of female steroid hormones --- p.5 / Chapter 1.2.2. --- Non-genomic actions of female steroid hormones --- p.7 / Chapter 1.2.3. --- Estrogen antagonists --- p.7 / Chapter 1.2.3.1. --- Classification of estrogen antagonists --- p.7 / Chapter 1.2.3.2. --- Mechanisms of estrogen antagonists --- p.9 / Chapter 1.3. --- Chronic (genomic) Effects of 17β-Estradiol and Progesterone --- p.10 / Chapter 1.3.1. --- Effects of lipid metabolism --- p.10 / Chapter 1.3.2. --- Effects on cell proliferation --- p.11 / Chapter 1.3.3. --- Effects on endothelial cells --- p.12 / Chapter 1.4. --- Acute Effects of 17β-Estradiol and Progesterone --- p.13 / Chapter 1.4.1. --- Role of endothelium in 17β-estradiol or progesterone Relaxation --- p.13 / Chapter 1.4.2. --- Involvement of plasma membrane estrogen receptors --- p.14 / Chapter 1.4.3. --- Role of Ca2+ and K+ channel in estrogen relaxation --- p.14 / Chapter 1.4.4. --- Interaction with vasoconstrictors --- p.15 / Chapter 1.4.5. --- Interaction with endothelium-dependent dilators --- p.16 / Chapter 1.4.6. --- Interaction with adrenergic response --- p.17 / Chapter 1.5. --- Clinical Studies --- p.19 / Chapter 1.6. --- Therapeutic Values of Estrogen and Progesterone --- p.20 / Chapter 1.7. --- Objectives of the Present Study --- p.22 / Chapter Chapter 2 --- Method and Materials / Chapter 2.1. --- Tissue Preparation --- p.25 / Chapter 2.1.1. --- "Preparation of the rat aorta, mesenteric artery and carotid Artery" --- p.25 / Chapter 2.1.2. --- Removal of the functional endothelium --- p.26 / Chapter 2.2. --- Organ Bath Set-up --- p.26 / Chapter 2.3. --- Force Measurement --- p.28 / Chapter 2.3.1. --- Vascular action of 17β-estradiol and progesterone --- p.29 / Chapter 2.3.1.1. --- Role of endothelium/nitric oxide in 17β-estradiol- or progesterone-induced relaxation --- p.29 / Chapter 2.3.1.2. --- Role of inducible nitric oxide in progesterone-induced relaxation --- p.30 / Chapter 2.3.1.3. --- Effect of estrogen receptor inhibitor on 17β-estradiol- induced relaxation --- p.30 / Chapter 2.3.1.4. --- Interaction between progesterone and 17β-estradiol --- p.31 / Chapter 2.3.1.5. --- Effect of 17β-estradiol on protein kinase C-mediated contraction --- p.31 / Chapter 2.3.1.6. --- Synergistic interaction between β-adrenoceptor agonists and 17β-estradiol --- p.32 / Chapter 2.4. --- Porcine Coronary Artery Experiments --- p.33 / Chapter 2.4.1. --- Vessel preparation --- p.33 / Chapter 2.4.2. --- Force measurement --- p.33 / Chapter 2.4.3. --- Experimental protocol --- p.34 / Chapter 2.4.3.1. --- Effect of physiological level of 17β-estradiol on β- adrenoceptor agonist-induced relaxation --- p.34 / Chapter 2.4.3.2. --- Effect of physiological level of 17β-estradiol on phosphodiesterase inhibitor-induced relaxation --- p.34 / Chapter 2.5. --- Ovariectomy --- p.35 / Chapter 2.5.1. --- Method of ovariectomy --- p.35 / Chapter 2.5.2. --- Preparation of blood vessels --- p.36 / Chapter 2.5.3. --- Experimental protocols --- p.38 / Chapter 2.5.3.1. --- Effect of ovariectomy on contractility of rat carotid arteries --- p.38 / Chapter 2.5.3.2. --- Effect of ovariectomy on relaxation of rat carotid arteries --- p.38 / Chapter 2.6. --- Chemicals and Solutions --- p.39 / Chapter 2.7. --- Statistical Analysis --- p.42 / Chapter Chapter 3 --- Results / Chapter 3.1. --- Role of Endothelium/Nitric Oxide in 17β-Estradiol- and Progesterone-induced Relaxations --- p.43 / Chapter 3.1.1. --- Relaxant response of 17β-estradiol --- p.43 / Chapter 3.1.2. --- Effects of inhibitors of nitric oxide activity on 17β- estradiol-induced relaxation --- p.46 / Chapter 3.1.3. --- Relaxant response of progesterone --- p.46 / Chapter 3.1.4. --- Effects of inhibitors of nitric oxide activity on progesterone-induced relaxation --- p.50 / Chapter 3.2. --- Effect of Estrogen Receptor Inhibitor on 17β-Estradiol- induced Relaxation --- p.56 / Chapter 3.3. --- Interaction between Progesterone and 17β-Estradiol --- p.56 / Chapter 3.4. --- Effect of Female Sex Steroid Hormones on Protein Kinase C-mediated Contraction --- p.59 / Chapter 3.4.1. --- Effect of 17β-estradiol on phorbol ester-induced contraction --- p.59 / Chapter 3.4.2. --- Effect of progesterone on phorbol ester-induced contraction --- p.59 / Chapter 3.5. --- Effects of β-adrenoceptor Agonists on 17β-Estradiol- induced Relaxations --- p.62 / Chapter 3.5.1. --- Effect of isoproterenol on 17β-estradiol-induced relaxation --- p.62 / Chapter 3.5.2. --- Role of endothelium/nitric oxide on the isoproterenol potentiation of 17β-estradiol-induced relaxation --- p.63 / Chapter 3.5.3. --- Role of cyclic AMP on isoproterenol-enhancement of 17β- estradiol-induced relaxation --- p.69 / Chapter 3.5.4. --- Effects of β-adrenoceptor antagonists --- p.69 / Chapter 3.6. --- Effects of Physiological Concentration of 17β-EstradioI onβ-adrenoceptor Agonists-induced Relaxationsin Porcine Coronary Artery --- p.77 / Chapter 3.6.1. --- Effect of 17β-estradiol on isoproterenol-induced relaxations --- p.77 / Chapter 3.6.2. --- Effect of 17β-estradiol on fenoterol-induced relaxations --- p.11 / Chapter 3.6.3. --- Effect of 17β-estradiol on dobutamine-induced relaxations --- p.81 / Chapter 3.6.4. --- Effect of 17β-estradiol on IBMX-induced relaxation --- p.86 / Chapter 3.7. --- Effect of Ovariectomy on the Vascualr Reactivity --- p.88 / Chapter 3.7.1. --- Effect of ovariectomy on the contractile activity of rat carotid artery --- p.88 / Chapter 3.7.1.1. --- Effect of ovariectomy on phenylephrine-induced contraction --- p.88 / Chapter 3.7.1.2. --- Effect of ovariectomy on U46619-induced contraction --- p.96 / Chapter 3.7.1.3. --- Effect of ovariectomy on high K+- induced contraction --- p.102 / Chapter 3.7.1.4. --- Effect of ovariectomy on acetylcholine-induced relaxation --- p.106 / Chapter Chapter 4 --- Discussions / Chapter 4.1. --- Role of Endothelium/Nitric oxide in 17β-Estradiol- and Progesterone-induced Relaxations --- p.110 / Chapter 4.2. --- Effect of Estrogen Receptor Inhibitor on 17β-Estradiol- induced Relaxation --- p.113 / Chapter 4.3. --- Interaction between Progesterone and 17β-Estradiol --- p.114 / Chapter 4.4. --- Effects of Female Sex Steroid Hormones on Protein Kinase C-mediated Contraction --- p.115 / Chapter 4.5. --- Effects of β-Adrenoceptor Agonists on 17β-Estradiol- induced Relaxations --- p.116 / Chapter 4.6. --- Effects of 17β-Estradiol on β-Adrenoceptor Agonists- induced Relaxations in Porcine Coronary Artery --- p.121 / Chapter 4.7. --- Effect of Ovariectomy on the Vascular Reactivity --- p.125 / Chapter 4.8. --- Conclusions --- p.129 / References --- p.131 / Publications --- p.145