鄭劍廷Chien, Chiang-Ting李孟哲Ly, Meng-Che2022-06-082027-01-142022-06-082022https://etds.lib.ntnu.edu.tw/thesis/detail/a776a049ccb28e776dda86bacb7fd5e3/http://rportal.lib.ntnu.edu.tw/handle/20.500.12235/117136膀胱是人體中重要器官之一，主要功能包含暫存尿液及排除含代謝廢物的尿液。隨著年齡增加，高齡人口表現膀胱過動症的比例也隨之遞增，考慮到生活品質降低和社會醫療成本增加，過動膀胱症是一個不可忽視的醫療議題。膀胱過動症為一種泌尿系統疾病，常見症狀如頻尿、尿急、儲尿功能下降，致病原因有很多，如糖尿病、下尿道阻塞、細菌感染以及尿路上皮受損，病 理 上 可 觀 察 到 逼 尿 肌 過度敏感 、 慢 性 發 炎 等 現 象 。 環 磷 醯 胺(Cyclophosphamide, CYP) 作為一種化療藥物，常在組織移植過程作為免疫抑制劑使用，在體內經過代謝後產生丙烯醛 (Acrolein)，經過腎臟濾出後順著尿液經過膀胱，將對膀胱造成損傷形成出血性膀胱炎，因此使用環磷醯胺作為誘導過動膀胱症的藥物。目前對於過動膀胱症的治療藥物多半伴隨不適的副作用，往往降低病患服藥意願，脂肪幹細胞為脂肪組織中具有自我修復與增殖的細胞群，許多研究注重脂肪幹細胞應用於活體中的作用以及培養脂肪幹細胞時分泌之微囊泡或胞外體應用於實驗研究中。因為自體細胞移植產生較少的免疫排斥、較高的組織相容性等。先前研究已各別使用脂肪幹細胞與脂肪幹細胞分泌之微囊泡治療膀胱缺血大鼠模式，能有效改善大鼠頻尿、缺血、發炎等。本研究欲探討脂肪幹細胞分泌之微囊泡應用於 CYP 誘導出血性膀胱炎中是否具抑制膀胱過度活化以及抗發炎效果。實驗結果顯示脂肪幹細胞分泌之微囊泡可下調減少第三型毒蕈鹼受體減少膀胱逼尿肌過度活化、減緩頻尿、過敏化的現象和以及下調減少 P2X7 受體表現量以及下游 Caspase 1、IL-1β等發炎路徑中相關分子的表現量，病理上觀察脂肪幹細胞所分泌之微囊泡可以降低免疫細胞浸潤現象、維持尿路上皮完整程度與抑制膀胱纖維化等結果。總結，脂肪幹細胞分泌之微囊泡可以改善CYP 引起之過動膀胱症。The urinary bladder plays an important role in human body. Its main functionsinclude temporary storage of urine and emptying of urine rich in metabolic waste.With the increase of age, the proportion of elderly people with overactive bladderalso increases. Considering the decline in the quality of life and the increase in social medical costs, overactive bladder is a medical issue that cannot be ignored.Overactive bladder is a urinary system disease, which can be caused by manyreasons, such as diabetes, lower urethral obstruction, bacterial infection, andurothelial damage. Pathologically, over-sensitivity of the detrusor muscle, decreased capacity of the bladder, frequent urination, and chronic inflammation can be observed. As a chemotherapeutic drug, Cyclophosphamide (CYP) is often used as an immunosuppressant during tissue transplantation. Acrolein, a metabolite of CYP, was produced and filtered out through kidneys. Ultimately, acrolein reaches urinary bladder via urine, causes damage to the bladder and forms hemorrhagic cystitis. In the present study, I use cyclophosphamide as an agent to induce overactive bladder disease.Most of the medial treatments for overactive bladder are accompanied byuncomfortable side effects, which often reduce the patient’s willingness to take the drug. Adipose stem cells are a group of cells that have self-repair and proliferation in adipose tissue. Many studies have focused on the role and culture of adipose stem cells in vivo and in vitro. Microvesicles or exosome secreted by adipose stem cells are used in experimental research. Due to autologous cell transplantation produces less immune rejection and higher histocompatibility. Our laboratory has successfully used adipose stem cells and adipose stem cells derived microvesicles to treat the bladder ischemia induced overactive bladder in the rat model. These results displayed that microvesicless treatment effectively improved urinary frequency ischemia, and inflammation in rats. This research intends to investigate whether microvesicles secreted by adipose stem cells can attenuate bladder overactivity and exert anti-inflammatory effects in CYP-induced hemorrhagic cystitis.Our experimental results show that the microvesicles secreted by adipose stemcells downregulate the type 3 muscarinic receptor, reduce the overactivation of thebladder detrusor, slow down the phenomenon of frequent urination, and reduceexpression of P2X7 receptors which involved downstream inflammatory pathwaysmolecule such as Caspase 1, and IL-1β. The pathologic results found that the thereduction of immune cell infiltration, the preservation of the integrity in theurothelium, and the inhibition of CYP-induced fibrosis in the bladder were discovered by the treatment of adipose stem cells derived microvesicles. Insummary, adipose stem cells derived microvesicles can ameliorate CYP-inducedbladder overactivity.環磷醯胺過動膀胱症脂肪幹細胞微囊泡毒蕈鹼受體嘌呤受體出血性膀胱炎cyclophosphamideoveractive bladderadipose stem cellsmicrovesiclesmuscarinic receptorspurinergic receptorshemorrhagic cystitis學術論文