氯胺酮是一種麻醉劑，也是一種濫用藥物。近年來，氯胺酮濫用增長迅速，在香港已經成爲第二大濫用藥物。氯胺酮的短期效果主要導致精神狀態改變，但對其長期效果還了解甚少。研究目的：本研究旨在探討長期使用氯胺酮對中樞神經系統，腎上腺，胰腺和膀胱的影響。研究方法：我們在氯胺酮濫用的動物模型中進行的行爲學，神經化學，組織學和分子生物學研究。在水迷宮中，對這些小鼠學習和記憶能力進行了評估。應用基因芯片評估中樞神經系統的基因表達變化。用聚合酶鏈式反應陣列研究神經遞質及其調控基因表達變化。通過實時定量聚合酶鏈式反應和免疫印迹法檢測-氨基丁酸受體和多巴胺相關基因的基因表達的變化。用酶聯免疫法測定多巴胺含量。用原位末端轉移酶標記技術染色（細胞凋亡），天狼星紅染色（纖維化），免疫組織化學（乳酸脫氫酶，酪氨酸羟化酶，多巴胺β羟化酶）研究腎上腺，胰腺，膀胱癌的組織學變化。研究結果：與對照組相比，氯胺酮組小鼠學習記憶能力下降。基因芯片結果顯示，110個基因表達上調和136個基因表達下調。基因本體分析表明，氯胺酮明顯影響神經遞質和受體的活性。特別地，4-氨基丁酸A受體5型亞基的mRNA和蛋白水平在前額皮層的顯著上調。聚合酶鏈式反應陣列結果表明，氯胺酮顯著改變-氨基丁酸受體，神經肽，多巴胺和膽鹼能系統基因表達。對多巴胺系統的研究結果顯示，中腦多巴胺含量上調，酪氨酸羟化酶的顯著增加。在腎上腺和胰腺，氯胺酮和氯胺酮加酒精組都未觀察到細胞凋亡增加，但是觀察到乳酸脫氫酶的陽性染色。此外，在腎上腺中發現酪氨酸羟化酶和多巴胺β羟化酶下調。在膀胱中，在肌肉層觀察到細胞凋亡和纖維化。結論：本研究的結果研究指出，長期使用氯胺酮能引起中樞神經系統異常的基因表達，還能導致腎上腺，胰腺，膀胱癌的病理性改變。這些結果爲氯胺酮濫用相關的健康風險評估提供了重要的信息。 / Ketamine is an anesthetic agent and a drug of abuse. In recent years, ketamine abuse has been increasing rapidly and it has become the second-most popular abusive drug in Hong Kong. While the acute effects of ketamine are mainly linked to altered mental status, the long-term effects of ketamine are poorly understood. Objectives: The present study was designed to investigate the long-term effects of ketamine on the CNS, adrenal, pancreas and urinary bladder. Methods: Behavioral, neurochemical, histological and molecular studies were performed in a ketamine abuse animal model. Learning and memory ability in these mice were assessed in a morris water maze. An Affymetrix Genechip study was performed to assess the global gene expression changes in the CNS and a PCR-array study focused on the neurotransmitters and regulators was also performed. Gene expression changes for gamma-aminobutyric acid (GABA) receptors and dopamine related genes were assay by real-time PCR and western blot. Dopamine contents were measured by ELISA. Histological changes in adrenal, pancreas and urinary bladder were examined by TUNEL staining (apoptosis), Sirius red staining (fibrosis), and immunohistochemistry. Results: Compared with saline controls, there was a decline in learning and memory performance in the ketamine-treated mice. Genechip results showed that 110 genes were up-regulated and 136 genes were down-regulated in ketamine group. An ontology analysis revealed the most significant effects of ketamine were on neurotransmitter and receptor activities. In particular, there was a significant up-regulation of both mRNA and protein levels of the alpha 5 subunit (Gabra5) of the GABAA receptors in the prefrontal cortex. Results from the PCR-array study revealed significant gene expression changes in the GABA receptors, neuropeptides, dopaminergic and cholinergic system following ketamine treatment. Studies on the DA system revealed significant increase of DA content and up-regulation of Tyrosine Hydroxylase (TH) in the midbrain. In the adrenal and pancreas, no obvious apoptosis was found while lactate dehydrogenase (LDH) positive staining was observed in both ketamine and ketamine plus alcohol treated groups. On top of these, downregulation of TH and DBH were observed. In the urinary bladder, apoptosis and fibrosis were observed in the muscular layer. Conclusion: The present study pointed out that long-term of ketamine use caused aberrant gene expression in the CNS and led to pathological changes in adrenal, pancreas and urinary bladder. These results have provided novel and important insights in evaluating the health risks in ketamine abusers. / Detailed summary in vernacular field only. / Tan, Sijie. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 138-154). / 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 --- p.I / 摘 要 --- p.III / List of abbreviations --- p.IV / Acknowledgements --- p.VI / Contents --- p.VII / Chapter Chapter 1 --- General introduction --- p.1 / Chapter 1.1 --- Ketamine and abuse --- p.1 / Chapter 1.2 --- Pharmacological effects of ketamine --- p.4 / Chapter 1.3 --- Effects of ketamine on the CNS --- p.6 / Chapter 1.4 --- GABA receptors --- p.9 / Chapter 1.5 --- Dopamine system in the CNS --- p.10 / Chapter 1.6 --- The modulation of dopaminergic neurons --- p.12 / Chapter 1.7 --- Toxic effects of ketamine on other organs --- p.14 / Chapter 1.8 --- Thesis outline --- p.17 / Chapter Chapter 2 --- Cognition and GABA receptor expression following long-term ketamine administration --- p.20 / Chapter 2.1 --- Introduction --- p.20 / Chapter 2.2 --- Materials and methods --- p.22 / Chapter 2.2.1 --- Animals and drug administrations --- p.22 / Chapter 2.2.3 --- Dosage Determination --- p.23 / Chapter 2.2.4 --- Morris water maze --- p.24 / Chapter 2.2.5 --- Brain tissue collection and RNA extraction --- p.25 / Chapter 2.2.6 --- Microarray analysis --- p.26 / Chapter 2.2.7 --- Quantitative real-time PCR --- p.27 / Chapter 2.2.8 --- Western blotting --- p.28 / Chapter 2.2.9 --- Statistical analysis --- p.29 / Chapter 2.3 --- Results --- p.29 / Chapter 2.3.1 --- Morris water maze --- p.29 / Chapter 2.3.2 --- Microarray analysis --- p.30 / Chapter 2.3.3 --- Quantitative real-time PCR --- p.30 / Chapter 2.3.4 --- Western blotting --- p.31 / Chapter 2.4 --- Discussion --- p.44 / Chapter Chapter 3 --- PCR-array gene expression profiling on the neurotransmitters following chronic ketamine administration --- p.49 / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Materials and methods --- p.50 / Chapter 3.3 --- Results --- p.51 / Chapter 3.4 --- Discussion --- p.63 / Chapter Chapter --- 4 Chronic ketamine administration modulates midbrain dopamine system in mice --- p.67 / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.2 --- Materials and methods --- p.69 / Chapter 4.2.1 --- Cell culture and ketamine treatment --- p.69 / Chapter 4.2.2 --- MTT assay --- p.70 / Chapter 4.2.3 --- Animals and ketamine administration --- p.70 / Chapter 4.2.4 --- Dopamine determination --- p.71 / Chapter 4.2.5 --- Real-time PCR --- p.72 / Chapter 4.2.6 --- Western blotting --- p.73 / Chapter 4.2.7 --- Immunohistchemistry --- p.74 / Chapter 4.2.8 --- Statistical analysis --- p.74 / Chapter 4.3 --- Results --- p.75 / Chapter 4.3.1 --- Effects of ketamine on dopamine concentrations in PC12 cells --- p.75 / Chapter 4.3.2 --- Long-term effects of ketamine on dopamine in mouse brain --- p.76 / Chapter 4.3.3 --- Effects of ketamine on mRNA levels of dopamine related genes --- p.76 / Chapter 4.3.4 --- Long-term effects of ketamine on BDNF protein levels in mouse brain --- p.77 / Chapter 4.3.5 --- Increased TH inmmureactive neurons in midbrain following 3 months ketamine treatment --- p.78 / Chapter 4.4 --- Discussion --- p.90 / Chapter Chapter 5 --- Chronic treatment of ketamine affects adrenal gland and pancreas --- p.95 / Chapter 5.1 --- Introduction --- p.95 / Chapter 5.2 --- Materials and methods --- p.95 / Chapter 5.2.1 --- Grouping of experimental animals and treatments --- p.95 / Chapter 5.2.2 --- Histological studies on pancreas and adrenal --- p.96 / Chapter 5.2.3 --- Immunohistochemistry on pancreas and adrenal --- p.97 / Chapter 5.2.4 --- TUNEL evaluation --- p.99 / Chapter 5.3 --- Results --- p.100 / Chapter 5.4 --- Discussion --- p.114 / Chapter Chapter 6 --- Ketamine Effects on the Urogenital System-Changes in the Urinary Bladder and Sperm Motility --- p.117 / Chapter 6.1 --- Introduction --- p.117 / Chapter 6.2 --- Materials and methods --- p.117 / Chapter 6.2.1 --- Studies of the Bladder --- p.117 / Chapter 6.2.2 --- Studies on Sperm Motility --- p.120 / Chapter 6.3 --- Results --- p.121 / Chapter 6.4 --- Discussion --- p.131 / Chapter Chapter 7 --- Conclusion --- p.134 / Chapter 7.1 --- Conclusion --- p.134 / Chapter 7.2 --- Future studies --- p.137 / Bibliography --- p.138
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_328237 |
| Date | January 2012 |
| Contributors | Tan, Sijie., Chinese University of Hong Kong Graduate School. Division of Biomedical Sciences. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | electronic resource, electronic resource, remote, 1 online resource (ix, 154 leaves) : ill. (some col.) |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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