- 學位論文
探討引發膀胱過動性的分子病理學機轉與治療策略
The molecular pathological mechanism and therapeutic modulation of urinary bladder hyperactivity
摘要
目的: 影響膀胱過動性的可能病理機轉十分多元，尿路上皮的完整性，聚葡萄糖胺(GAG)層的屏障作用，粒線體及內質網(ER)因代謝、老化、荷爾蒙、動脈硬化等因素受到自由基傷害增加氧化壓力及發炎反應等等狀況，均會影響膀胱功能及造成解尿障礙，因此本研究主要以一系列實驗研究膀胱過動性的病理機轉，並應用在臨床上治療策略的選擇以動物實驗模式，來針對不同病因所引發的過動性膀胱做詳細的分析並嘗試解釋可能的致病模式及治療機轉。 方法: 泌尿上皮完整性受到K它命破壞後會引起發炎反應並造成頻尿、急尿膀胱過動症狀，以玻尿酸(hyaluronic acid , HA)灌注治療實驗，觀察K它命腹膜注射雌性大白鼠解尿行為及膀胱壓力，評估膀胱完整性受K它命腹膜注射後是否影響膀胱過動性，以免疫染色的方法，檢視細胞病理型態變化，試驗玻尿酸(HA)灌注促進膀胱修補的細胞學機轉。也以飲食引起高血糖高血脂代謝障礙之大白鼠模擬並觀察對膀胱過動性的影響，並評估若合併卵巢切除之雌性大白鼠給予不同濃度的兒茶素(epigallo-catechin-3-gallate, EGCG)注射後其氧化壓力及細胞凋亡相關蛋白質的表現以了解由代謝症候群及卵巢荷爾蒙缺損所引發之過動性膀胱其內質網細胞凋亡路徑 和粒線體呼吸鏈作用酶對於代謝症候群(MetS)和卵巢功能缺損(ovarian hormone deficiency )的細胞學機轉及治療策略。 結果: 玻尿酸(HA)的研究證實，K它命有正向調節(up-regulation)性的刺激會增進環氧合酶2(COX-2) 和轉化生長因子β1的蛋白質表現，而從膀胱灌注玻尿酸會降低環氧合酶2(COX-2) 和轉化生長因子β1的表現，可以調節膀胱的過動性。在K它命注射實驗組的尿路上皮層中，細胞增生蛋白Ki67、分化標記uroplakin III及細胞表面抗原44(CD44)的蛋白質表現均明顯較控制組低，以玻尿酸進行膀胱灌注治療，能引發尿路上皮層細胞的增生(proliferation)與分化(differentiation)，因而刺激尿路上皮黏膜的再生。兒茶素EGCG的動物實驗中則證實，卵巢切除造成的卵巢功能缺損狀態和高糖高脂飲食所致代謝症候群大白鼠會誘導GRP78、 CHOP和caspase-12的表現增加，增加膀胱組織的細胞凋亡反應，並刺激膀胱組織內細胞凋亡相關蛋白的表現，活化粒腺體相關細胞凋亡路徑之分子訊號，對於大白鼠的膀胱組織造成極大的氧化傷害，細胞藉由兒茶素EGCG的抗氧化作用可以調節粒線體的功能，緩解代謝症候群(MetS)和卵巢功能缺損(ovarian hormone deficiency )的影響。 結論: 玻尿酸動物實驗發現玻尿酸藉由活化尿路上皮相關黏著蛋白E-cadherin、細胞增生標記蛋白Ki67以及刺激尿路上皮分化標記uroplakin III，得以修補重建膀胱尿路上皮緩解K它命所致的膀胱過動。而兒茶素EGCG動物實驗則顯示，代謝障礙和卵巢荷爾蒙缺損會藉由粒線體與內質網路徑增加氧化壓力和細胞凋亡反應。兒茶素EGCG會減弱氧化壓力並調控粒線體與內質網細胞凋亡路徑的分子訊號，幫助細胞抵抗氧化傷害，可能是治療下泌尿道膀胱過動症狀的新思維與策略。
並列摘要
Objectives: The etiology of bladder hyperactivity is multifactorial related to urothelial integrity, oxidative damage to mitochondria and endoplasic reticulum (ER) in response to metabolic, aging, hormonal and arthrosclerotic conditions. Free radical injury to excite oxidative stress and inflammatory reactions could be pivotal to bladder. A series of animal experiments were designed to investigate the molecular pathological mechanism and therapeutic modulation of urinary bladder hyperactivity. Methodszz: Chemical ketamine destroyed the urethelial integrity resulting in bladder hyperactivity symptoms such as frequency and urgency. In the hyaluronic acid (HA) instillation study, female Sprague-Dawley rats intraperitoneally injected with ketamine. By immunofluorescence analysis, the cellular pathological behaviors were observed to elucidate the mechanisms of bladder repair after HA instillation. However, in the epigallocatechin-3-gallate (EGCG) study attempted to clarify the mechanisms of EGCG modulating oxidative stress in metabolic syndrome (MetS) and ovarian hormone deficiency-associated bladder hyperactivity by examing the expressions of oxidative markers and mitochondria respiratory enzyme complexes in the bladders. Results: In the HA study, the ketamine-treated rats exhibited significant bladder hyperactivity, the expressions of cyclooxygenase 2(COX-2) and TGF β1 were up-regulated by ketamine. Moreover, HA instillation down regulated the expressions of cyclooxygenase 2(COX-2) and TGF β1, ameliorated bladder hyperactivity, and lessened interstitial fibrosis. In the urothelial layer of ketamine-treated bladder, the expressions of Ki67, uroplakin III, and CD44 were decreased than control. After HA instillation, the regeneration of bladder were excited in the urothelium. In the EGCG study, the rats fed with high fat high sugar (HFHS) diet plus bilaterally ovariectomy would deteriorate the changes of metabolic syndrome (MetS) and bladder hyperactivity. The expressions of GRP78, CHOP, caspase-12, mitochondrial apopototic signals were motivated with apoptotic reactions and proteins in the metabolic syndrome and ovarian hormone deficiency rats. EGCG could modulate oxidative stress through mitochondrial and ER apoptotic pathways to ameliorate the bladder hyperactivity associated with MetS and ovarian hormone deficiency. Conclusionsz: HA could modulate E-cadherin, Ki67 and uroplakin III to restore urothelial integrity and bladder hyperactivity induced by ketamine. The metabolic syndrome and ovarian hormone deficiency incited the oxidative stress and apoptosis through mitochondrial and ER apoptotic pathways. Antioxidant EGCG offered a beneficial effect on lessening oxidative damages through mitochondria and ER-mediated pathways. A better understanding of the relevant pathophysiology of bladder hyperactivity could lead to noval therapeutic modulations.
參考文獻
延伸閱讀
- 趙惠如（2020）。膀胱過動症患者對膀胱狀況之感受與症狀困擾相關性探討-以問題取向因應行為和情緒反應為中介變項之比較〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU202001914
- Peng, C. H. (2010). 間質性膀胱炎之膀胱內藥物治療及症狀影響因子評估 [master's thesis, National Taiwan University]. Airiti Library. https://doi.org/10.6342/NTU.2010.01938
- Lin, Y. H., Huang, H. C., Chiang, B. J., Wang, M. H., & Liao, C. H. (2018). 膀胱過動症患者合併尿失禁病人施行體外電慈刺激：單一機構之研究經驗分享. 輔仁醫學期刊, 16(2), 57-65. https://doi.org/10.3966/181020932018061602001
- Cheung, T. (1996). A study of urinary-type plasminogen activators in biological fluids [doctoral dissertation, The University of Hong Kong]. Airiti Library. https://www.airitilibrary.com/Article/Detail?DocID=U0029-1812201200007383
- Neuhaus, J., Schwalenberg, T., Horn, L. C., Alexander, H., & Stolzenburg, J. U. (2011). New Aspects in the Differential Diagnosis and Therapy of Bladder Pain Syndrome/Interstitial Cystitis. Advances in Urology, 2011(), 151-159. https://doi.org/10.1155/2011/639479