劉馨慈副研究員

藉由蛋白質分析技術如酵素接合免疫分析法(ELISA)、免疫組織化學染色(Immunohistochemistry)及分子生物技術如即時聚合酶鏈鎖反應(real-time PCR)等，本研究室檢測病患血清、尿液及組織中感覺神經蛋白質—如神經生長因子(NGF)、感覺神經受器--(TRPV1、P2X3)、發炎反應相關物質--前列腺素E2 (PGE2)及氧化壓力相關蛋白質，於膀胱過動症病患之表現狀態。進而將這些研究數據進行統計及資料分析後，將病人與對照組的表現量相互比較，以厘清膀胱過動症的致病原因。此外本研究室亦取膀胱過動症病患於肉毒桿菌毒素A治療前後的膀胱組織，分析其治療前後之功能性蛋白質變化情形。從而瞭解肉毒桿菌毒素A的療效機轉。

目前的研究結果顯示:(1)膀胱過動症病人, 其膀胱組織中肥大細胞數量顯著增多，合併血清及尿液中神經生長因子及發炎相關物質明顯上升，推論部份原發性之膀胱過動症可能與膀胱局部或全身慢性發炎有關。(2)抗膽鹼藥物治療無效之病人，其膀胱黏膜組織中P2X3與PGP9.5蛋白質的含量兩者皆明顯較對照組高。由此推論，膀胱內感覺神經纖維與分布於感覺神經上的感覺神經受器P2X3的異常增生可能是造成這類膀胱過動症的致病原因之一。近年來我們亦發現缺血再灌流所產生的氧化壓力在膀胱過動症的致病機轉上扮演一定的角色。

研究室未來的目標，將以大鼠和基因轉殖小鼠的動物轉譯模式，觀察尿路感染及氧化壓力對膀胱及身體其他器官所造成的影響。進而以目前臨床常用之抗氧化藥物治療，觀察藥物治療前後之分子病理變化。由此了解氧化壓力所造成的疾病機轉及抗氧化藥物的療效。期望本實驗室之研究成果，能提供臨床醫師於相關疾病診斷及治療時的一個客觀輔助參考。

The main research focus in the laboratory is the investigation of the pathophysiological mechanisms underlying overactive bladder (OAB) syndrome. Our interest in this topic developed through elucidation of the roles of sensory nerve proteins (e.g., nerve growth factor; NGF), sensory nerve receptors (e.g., TRPV1 and P2X3), inflammation-related substances (e.g., prostaglandin E2; PGE2), and oxidative stress-related proteins in the bladders of patients with OAB. We use protein analysis techniques, such as enzyme-linked immunoassay (ELISA) and immunohistochemistry, and molecular biology techniques, such as real-time polymerase chain reaction (real-time PCR), to analyze the regulation and expression of these proteins in the serum, urine and tissues of OAB patients. Using statistical analyses, we compare the protein expressions between OAB patients and control subjects to identify the pathogenesis of OAB. In addition, in order to understand the therapeutic mechanism of onabotulinumtoxinA (BoNT-A) therapy, our laboratory also investigates the changes of bladder functional proteins in the tissue of patients before and after BoNT-A treatment.

The results of our current study indicated the following: (1) increased numbers of mast cells in the bladder and elevated levels of serum and urinary NGF and pro-inflammation proteins, which suggested involvement of local or systemic inflammation in certain OAB patients; (2) elevation of P2X3 and PGP9.5 expressions in the bladder mucosa of patients refractory to antimuscarinic drugs, which implied that an abnormally increased level of the sensory receptor P2X3 is one part of the pathogenesis of the disease.

In recent years, we found that oxidative stress caused by ischemia and reperfusion also plays an important role in several diseases. Therefore, future goals of our laboratory include the use of rat and transgenic mouse models to study the effects of oxidative stress on the bladder and other organs, and the application of antioxidant drugs in current clinical use for the treatment of other diseases to explore whether the drugs cause molecular pathological changes in the bladders of patients. By comparison with clinical specimens from patients, our study combined basic research and clinical pathologic analysis, and the findings provide an objective reference for clinicians regarding the diagnosis and treatment of diseases. 

1.

Liu HT, Kuo HC: Expressions of urothelial functional proteins in idiopathic detrusor overactivity patients refractory to antimuscarinic therapy with different urodynamic characteristics. Neurourol Urodyn, 2016, Sep 21. doi: 10.1002/nau.23138. [Epub ahead of print].(SCI)

2.

Chuang FC, Liu HT, Kuo HC: Lower Levels of Urinary Nerve Growth Factor Might Predict Recurrent Urinary Tract Infections in Women. International Neurourology Journal, 2016, 20(1): 33-39.(SCI)

3.

Liu HT, Chen SH, Chancellor MB, Kuo HC: Presence of Cleaved Synaptosomal-Associated Protein-25 and Decrease of Purinergic Receptors P2X3 in the Bladder Urothelium Influence Efficacy of Botulinum Toxin Treatment for Overactive Bladder Syndrome. PLoS One, 2015, 4;10(8):e0134803. doi: 10.1371/journal.pone.0134803. (SCI)

Liu HT, Kuo HC: Biomarkers for patients with interstitial cystitis/bladder pain syndrome.  Urological Science, 2015, 26(4): 225-229. (Review)

Liu HT, Jiang YH, Kuo HC: Alteration of Urothelial Inflammation, Apoptosis, and Junction Protein in Patients with Various Bladder Conditions and Storage Bladder Symptoms Suggest Common Pathway Involved in Underlying Pathophysiology. LUTS-Lower Urinary Tract Symptoms, 2015, 7(2): 102-107.(SCI)

Jiang YH, Liu HT, Kuo HC: Urothelial dysfunction and chronic inflammation in patients with spinal cord injuries at different levels and correlation with urodynamic findings. Neurourol Urodyn, 2015, 34: 757-762.(SCI)

Jiang YH, Liu HT, Kuo HC: Decrease of Urinary Nerve Growth Factor but Not Brain-Derived Neurotrophic Factor in Patients with Interstitial Cystitis/Bladder Pain Syndrome Treated with Hyaluronic Acid. PLoS One, 2014.9(3): p. e91609(SCI)

Kuo HC, Liu HT,Chuang YC, Birder LA, Chancellor, MB: Pilot study of liposome-encapsulated onabotulinumtoxina for patients with overactive bladder: a single-center study. Eur Urol, 2014,65(6): 1117-1124(SCI)

Chuang FC, Liu HT, Wang LY, Kuo HC: Overactive bladder changes with time: a 5-year longitudinal followup of changes in overactive bladder symptoms, urodynamic studies and urinary nerve growth factor levels. J Urol, 2014,192(2): 458-463.(SCI)

Kuo HC , Liu HT, Shie JH: Potential urine and serum biomarkers for patients with overactive bladder and interstitial cystitis/bladder pain syndrome. Tzu Chi Medical Journal 2013; 25:13-18.

Liu HT, Jiang YH, Kuo HC: Increased Serum Adipokines Implicate Chronic Inflammation in the Pathogenesis of Overactive Bladder Syndrome Refractory to Antimuscarinic Therapy. PLoS One, 2013;8(10): p. e76706(SCI)

Jiang YH, Peng CH, Liu HT, Kuo HC: Increased Pro-Inflammatory Cytokines, C-Reactive Protein and Nerve Growth Factor Expressions in Serum of Patients with Interstitial Cystitis/Bladder Pain Syndrome. PLoS One, 2013;8(10): p. e76779(SCI)

ChenYR, Huang HB, Lo CJ, Wang CC, Ho LK, Liu HT, Shiao MS, Lin DT, Chen YC: Effect of Alanine Replacement of L17 and F19 on the Aggregation and Neurotoxicity of Arctic-Type Ab40. PLoS One, 2013.8(4): p. e61874 (SCI)