臺灣博碩士論文加值系統

糖尿病導致的膀胱增大現象，肇因於多尿症引起的慢性擴張作用。而對於糖尿病引起尿量增加，其調控膀胱平滑肌的作用機轉，目前仍不清楚。我們最近的研究發現，以腹腔注射方式給予實驗動物STZ誘發糖尿病產生的3-7天，除了血糖增高；膀胱重量增加外，亦發現 nNOS蛋白表現量也增加；且會誘發IL-6 mRNA表現，及促進IGFBP2蛋白量增加；而細胞外訊息調控激素ERK1及ERK2亦會被磷酸化。在我們使用STZ誘發實驗動物產生糖尿病的實驗模式中發現，一氧化氮合成酵素抑制劑-L-NAME，具有抑制IL-6 mRNA 及IGFBP2蛋白表現的能力，並且也會抑制ERK蛋白磷酸化。此外，anti-IL-6抗體同樣地也具有拮抗IGFBP2蛋白表現，及抑制ERK蛋白磷酸化的能力，並且會抑制膀胱肥大現象。再者，我們也發現有三種型態的PKC蛋白會由細胞質轉移至細胞膜；而其膀胱組織的收縮情形亦較血糖正常組劇烈。由上述的結果顯示：糖尿病狀態下，造成實驗老鼠膀胱組織的肥大現象，一氧化氮-NO及細胞激素-IL-6在這過程中扮演著重要的角色。
內毒素LPS，具有誘導發炎反應產生，而造成敗血症的特性。過去的研究報告指出，無論是以腹腔注射的方式給予LPS；或將LPS直接注射到膀胱中，皆發現在實驗大鼠的膀胱組織中，LPS會誘發iNOS表現。然而，iNOS在膀胱功能上所扮演的角色目前仍不清楚。我們最近的研究發現，給予實驗小鼠LPS後的3-24小時，在其膀胱組織中會誘發IL-6 mRNA、iNOS mRNA及iNOS蛋白的表現。此外，PKC在調控膀胱肌肉收縮功能上亦扮演著重要的角色。而LPS處理後，在其膀胱組織中也會促使三種型態的PKC蛋白-alpha、beta及gamma，由細胞質轉移至細胞膜。若同時給予具有選擇性的iNOS抑制劑-AG或L-NIL，則可拮抗此現象。由上述的結果顯示：受到內毒素感染的實驗小鼠，其膀胱收縮功能的改變與PKC蛋白及NO有著密不可分的關係。
綜合以上的結果：無論是在內毒素血症；或早期糖尿病狀態下，NO與IL-6在膀胱組織中，具有調控細胞生長及影響膀胱收縮等功能。

During diabetes, the urinary bladder is markedly enlarged, a result of chronic distension caused by polyuria. The mechanisms regulating adaptation of urinary bladder smooth muscle to the increased diuresis poorly understood. In the present study, we found that after 3-7 days of injection of streptozotocin (STZ), the increase of blood glucose, urinary bladder weight, expression of neuronal type-nitric oxide synthase (nNOS) protein and IL-6 mRNA, insulin-like growth factor-binding protein-2 (IGFBP-2), and phosphorylation of extracellular signal-regulated kinases (ERK 1 & 2), induce diabetes. The NOS inhibitor-L-NAME was capable of inhibiting the expressions of IL-6 mRNA and IGFBP-2 and phosphorylation of ERK induced by STZ-diabetes. Moreover, the anti-IL-6 antibody could also inhibit the IGFBP-2 expression, phosphorylation of ERK and hypertrophying bladder induced by STZ-diabetes. In addition, we also found that the translocation of conventional PKC isoforms from the cytosolic to the particulate fractions and the electrical field stimulation-induced muscle contraction in urinary bladder was enhanced. These results indicate that NO and IL-6 may play a important role in the urinary bladder hypertrophy in the diabetic mouse.
On the other hand, systemic bacterial lipopolysaccharide (LPS) induce inflammatory responses characteristic of sepsis. Both intraperitoneal injection and bladder instillation of LPS has been shown to induce the expression of inducing inducible nitric oxide synthase (iNOS) in rat urinary bladder. However, the role of iNOS induction in the function of urinary bladder is still unknown. In the present study, we found that a significant induction of IL-6 and iNOS mRNA and iNOS protein expression in the urinary bladder 3-24 hours after the application of LPS to mouse. In addition, protein kinase C (PKC) plays a key role in modulating muscle contraction of the mouse urinary bladder muscle. The translocation of the PKC alpha、beta and gamma isoforms from the cytosolic to the particulate fractions was found in LPS-treated mouse urinary bladder. The in vivo effects of LPS on the mouse urinary bladder could be prevented by the administration of AG and L-NIL (iNOS inhibitors). These results indicate that the changes of conventional PKC isoforms in urinary bladder may be related to the NO-mediated effects on contractions of urinary bladder in endotoxemic mice.
All of these results suggest that NO and IL-6 may be involved in the regulation of the cell growth and functions of urinary bladder during endotoxemic and early stage of diabetic conditions.

中文摘要------------------------------------------------1
英文摘要------------------------------------------------3
緒 論------------------------------------------------5
材料與方法----------------------------------------------9
結 果-----------------------------------------------23
討 論-----------------------------------------------28
參考文獻-----------------------------------------------37
圖 表-----------------------------------------------49

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