2015-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/651194摘要：膀胱出口阻塞 (bladder outlet obstruction; BOO)與許多泌尿科的疾病有關。過去的研究顯示膀胱出口阻塞造成膀胱傷害的機轉是因為過度的膀胱張力會導致缺氧（hypoxia），而引發一連串的細胞傷害，最終將造成膀胱組織學型態及功能的變化。這種因為缺血造成細胞傷害的相關機轉包括未摺疊蛋白反應（unfolded protein response；UPR）、內質網壓力（endoplasmic reticulum stress；ER stress）、壓力相 關 蛋 白 的 活 化 (heat shock proteins, c-Jun, heat-shock proteins, phospho-SAPK/JNK, phospho-c-Jun, phospho-AMPK) 、 mammalian target of rapamycin (mTOR)、氮化合物(nitrogen species; iNOS)及一氧化氮（nitric oxide; NO）、 細胞自噬作用（autophagy）、血管新生作用 (angiogenesis)等。目前臨床上仍無可用之藥物減輕膀胱傷害。 我們在過去的研究中已經發現，內質網壓力確實參與在其中，且利用 GRP78 inhibitor (epigallocatechin gallate; EGCG) 能抑制內質網壓力造成的膀胱傷害。尤其美國食品藥物管理局在 2011 年核准了第 5 型磷酸二脂酶抑制劑 (phosphodiesterase; PDE5 inhibitors) tadalafil治療前列腺增生相關的下泌尿到症狀, 這些機轉在膀胱出口阻塞造成的膀胱功能變化所扮演的角色，仍非常值得研究，也非常有潛力能發展出新的治療模式。 我們設計此一研究，主要目的在探討在膀胱出口阻塞造成膀胱傷害之機轉，針對這些尚未清楚的細胞傷害或保護機轉，找尋新的治療藥物並發展新的治療模式。 此為三年計畫 第一年 我們已經建立了兩種膀胱出口阻塞造成膀胱傷害之動物模式 :部分阻塞（partial bladder outlet obstruction; pBOO）及 bladder overdistension （BO），可用以探討造成膀胱出口阻塞造成膀胱傷害之機轉，我們先證實在不同的膀胱出口阻塞的時間點，包括 pBOO的急性期（48hr）,亞急性期（30 days）,慢性期（90 days）及 BO 48hr 時有關缺氧（hypoxia）及未摺疊蛋白（unfolded protein response）堆積的情形，並利用活體測量其膀胱功能變化。 第二年 根據第一年研究發現之結果，我們將進一步探討包括內質網壓力（ER stress）、壓力相關蛋白的活化 （stress-related prtoeins）、氮化合物(nitrogen species)及 NO、m-TOR、細胞自噬作用（autophagy）、血管新生作用 (angiogenesis)，釐清其在兩種膀胱出口阻塞模式中扮演的角色 （保護 vs. 傷害）。 第三年 利用調控這些細胞生理反應機轉，利用藥物如 m-TOR inhibitor (rapamycin), ER stress inhibitor (salubrinol), angiogenesis inducer (Okadaic acid)，PDE5 inhibitor (tadalafil), NOS inhibitor (L-NAME) 來評估藥物在兩種膀胱出口阻塞的模式中之治療效果，並利用活體測量其膀胱功能之改善情況，以證實此藥物之效果。在慢性期（>90 days），能否預防膀胱組織之纖維化(fibrosis)。 吾人相信，本研究將有助於了解膀胱出口阻塞之細胞傷害機轉，並藉此能找到有效之藥物，發展新的治療模式，日後能進一步應用於臨床。<br> Abstract: Bladder outlet obstruction (BOO) is a common outcomes of many urological diseases. However, the underlying mechanism of BOO-induced bladder injury remains unclear. Several previous reports suggested that excessive distension of bladder caused hypoxia, which is implicated in structural and functional alterations of the urinary bladder. Cells respond to hypoxia and ER stress through the activation of unfolded protein response (UPR), which is an adaptive response to increase cell survival during stressful condition. Meanwhile, autophagy, angiogenesis, m-TOR, stress-related protein activation (heat shock proteins, c-Jun, heat-shock proteins, phospho-SAPK/JNK, phospho-c-Jun, phospho-AMPK) and NO production are also involved in the process. What are the roles of these mechanisms play in BOO-induced bladder injury still remains elusive. Our previous study has proved that ER stress plays an important role in the process of BOO-induced bladder injury. Most important of all, we found GRP78 inhibitor, (epigallocatechin gallate; EGCG) could alleviate BOO-induced bladder injury by suppressing ER stress. In 2011, U.S. FDA approved the indications of tadalafil (phosphodiesterase; PDE5 inhibitors) in treating BPH-related lower urinary tract symptoms. These cellular responded to hypoxia and unfolded protein accumulation after BOO have potential to develop novel target for the treatment of BOO-induced bladder injury. In this study, we aim to delineate this important issue. We design the 3-year study In the first year We will take advantage of two animal models that we have established (partial bladder outlet obstruction; pBOO and bladder overdistension). We will investigate the role of hypoxia and UPR play in BOO-induced bladder injury at different time points---pBOO (acute stage<48 hr, sub-acute, 30 days, chronic 90 days) and bladder overdistension (48 hr). Also we will examine the bladder function by using in vivo cystometry. In the second year Based on the findings in the first year, we aim to investigate the roles of hypoxia and UPR related cellular response such as autophagy, angiogenesis, m-TOR, stress-related protein activation and NO production on BOO-induced bladder injury by using the two animal models respectively. In the third year By modulating these novel cellular responses, we will select some candidate drugs (m-TOR inhibitor (rapamycin), ER stress inhibitor (salubrinol), angiogenesis inducer (Okadaic acid)，PDE5 inhibitor (tadalafil), NOS inhibitor (L-NAME)) to examine the protective effects of these candidate drugs on BOO-induced bladder injury. Not only will we evaluate the histologic change and apoptotic effects, but also the detrusor function by in vivo cystometry. Additionally, we will validate the therapeutic mechanisms of these candidate drugs. We are confident that we will obtain the promising results, which will have important implications for the development of novel therapeutic strategies to prevent or alleviate bladder damage resulting from BOO.膀胱出口阻塞未摺疊蛋白反應內質網壓力一氧化氮細胞自噬作用血 管新生作用細胞凋亡bladder outlet obstruction (BOO)hypoxiaendoplasmic reticulum stressautophagyangiogenesisunfolded protein responseapoptosis