本文選題：膀胱過(guò)度活動(dòng)癥 + 逼尿肌細(xì)胞��； 參考：《第三軍醫(yī)大學(xué)》2014年碩士論文

【摘要】：目的： 膀胱過(guò)度活動(dòng)癥（Overactive bladder，OAB）是一種常見(jiàn)的泌尿外科疾病，其定義為：OAB是一種以尿急癥狀為特征的癥候群，常常伴有尿頻和夜尿癥狀，可伴或不不伴有急迫性尿失禁，，OAB可獨(dú)立發(fā)生，也可與其他泌尿系統(tǒng)疾�。ㄈ缒蚵犯腥�、老年性排尿異常、神經(jīng)源性膀胱、下尿路梗阻、間質(zhì)性膀胱炎、下尿路腫瘤、泌尿系結(jié)石、不明原因的男女性尿頻）相伴發(fā)生。OAB病因不明，目前存在神經(jīng)源性和肌源性兩種學(xué)說(shuō)，前者被廣泛接納但仍有不能解釋的疑問(wèn)，后者在近年來(lái)被廣泛關(guān)注。OAB的主要病理生理學(xué)改變是逼尿肌過(guò)度活動(dòng)（detrusor overactivity， DO）。 鈣離子（Ca2+）是細(xì)胞內(nèi)一種重要的無(wú)機(jī)陽(yáng)離子，Ca2+參與了細(xì)胞的許多生理活動(dòng)以及生理功能，在這其中，平滑肌的收縮性和興奮性的調(diào)節(jié)是一項(xiàng)重要作用。在鈣離子通道中，鈣庫(kù)調(diào)控的鈣離子通道（Store Operated Calcium Channels， SOCCs）參與了血管平滑肌、呼吸道平滑肌的收縮和興奮的調(diào)控，進(jìn)過(guò)論證，我們推測(cè)SOCCs可能在膀胱逼尿肌細(xì)胞的興奮性和收縮性的調(diào)節(jié)中發(fā)揮了重要作用，但是目前缺乏相關(guān)的證據(jù)。 因此，我們通過(guò)研究SOCCs在DO時(shí)的SD大鼠膀胱逼尿肌中的功能改變，探索SOCCs在OAB發(fā)生的機(jī)制和病理生理改變中的作用。 方法： 1.建立DO模型：取60只體質(zhì)量為180-200g的雌性SD大鼠，其中30只作為對(duì)照組，其余30只行下尿路部分梗阻（partial bladder outlet obstruction，PBOO）手術(shù)，術(shù)后6周通過(guò)膀胱測(cè)壓法鑒定，選取其中出現(xiàn)不穩(wěn)定收縮的，即為DO模型，組成DO組； 2.膀胱逼尿肌離體肌條實(shí)驗(yàn)：利用離體肌條牽拉實(shí)驗(yàn)，采用SOCCs激動(dòng)劑CPA和抑制劑SKF-96365干預(yù)，觀察SOCCs對(duì)膀胱逼尿肌離體肌條自發(fā)收縮情況（包括收縮頻率和收縮幅度）的影響，并對(duì)兩組的結(jié)果進(jìn)行統(tǒng)計(jì)學(xué)比較； 3.全細(xì)胞膜片鉗實(shí)驗(yàn)：通過(guò)2步法急性酶分離法分離得到膀胱逼尿肌細(xì)胞，分離得到的逼尿肌細(xì)胞采用全細(xì)胞膜片鉗技術(shù)，觀察其SOCCs電流，并用其激動(dòng)劑CPA和和抑制劑SKF-96365進(jìn)行干預(yù)，觀察SOCCs電流的變化情況，并對(duì)兩組的結(jié)果進(jìn)行統(tǒng)計(jì)學(xué)比較； 4.鈣負(fù)載實(shí)驗(yàn)：通過(guò)1步法急性酶分離法分離的膀胱逼尿肌細(xì)胞，培養(yǎng)24h后，用Fluo-4AM熒光染色，用激光共聚焦顯微鏡觀察逼尿肌細(xì)胞內(nèi)的鈣離子熒光，用CPA和SKF-96365進(jìn)行干預(yù)，觀察細(xì)胞內(nèi)鈣離子熒光變化情況，并對(duì)兩組的結(jié)果進(jìn)行統(tǒng)計(jì)學(xué)比較； 結(jié)果： 1. DO模型建立成功，通過(guò)膀胱測(cè)壓，30只進(jìn)行PBOO手術(shù)的雌性SD大鼠有25只出現(xiàn)不穩(wěn)定收縮，成為DO模型大鼠； 2.膀胱逼尿肌離體肌條實(shí)驗(yàn)提示，采用CPA激活SOCCs后，離體逼尿肌肌條自發(fā)收縮頻率加快，幅度降低，采用SKF-96365抑制SOCCs后，離體逼尿肌肌條自發(fā)收縮頻率減慢，幅度回升，DO組與對(duì)照組相比，其自發(fā)收縮頻率的變化有顯著差異（P0.05），而收縮幅度的變化差異不顯著； 3.全細(xì)胞膜片鉗實(shí)驗(yàn)提示，成功分離大鼠膀胱逼尿肌細(xì)胞，并記錄到CPA可激發(fā)SOCCs電流，SKF-96365可使SOCCs電流降低，兩組組相比，DO組在+20mv和-80mv的電流密度顯著高于對(duì)照組（P0.05）; 4.鈣負(fù)載實(shí)驗(yàn)提示：成功培養(yǎng)了原代膀胱逼尿肌細(xì)胞，利用激光共聚焦顯微鏡觀察可知CPA可增加細(xì)胞內(nèi)的鈣離子濃度[Ca2+]i，SKF-96365可降低細(xì)胞內(nèi)鈣離子濃度[Ca2+]i，兩組相比較，DO組的[Ca2+]i顯著高于對(duì)照組（P0.05）。 結(jié)論： 1. SOCCs參與調(diào)控了膀胱逼尿肌的收縮性和興奮性； 2. DO時(shí)，SOCCs活性發(fā)生改變； 3. SOCCs在DO時(shí)活性增強(qiáng)，為OAB病因及病理生理學(xué)研究提供了新的思路。
[Abstract]:Objective:
Overactive bladder (OAB) is a common disease in the Department of urology. Its definition is that OAB is a symptom characterized by urgent urination, often accompanied by frequency of urination and nocturia, with or without urgent incontinence, OAB can occur independently, and can also be associated with other urinary tract diseases (such as urinary tract infection, senile sex). Abnormal urination, neurogenic bladder, lower urinary tract obstruction, interstitial cystitis, lower urinary tract tumor, urinary calculi, unexplained frequency of male and female urinary frequency) associated with unknown.OAB etiology, there are two doctrines of neurogenic and muscular origin, the former is widely accepted but still have unexplained questions, the latter has been widely concerned in.OAB in recent years. The main pathophysiological change is detrusor overactivity (DO).
Calcium ion (Ca2+) is an important inorganic cation in cells. Ca2+ participates in many physiological and physiological functions of cells. In this, the regulation of contractility and excitability of smooth muscle is an important role. In calcium channel, calcium ion channel (Store Operated Calcium Channels, SOCCs) participates in blood in calcium channel. The regulation of smooth muscle, contraction and excitement of the smooth muscle of the respiratory tract has been demonstrated. We speculate that SOCCs may play an important role in the regulation of excitability and contractility of the bladder detrusor cells, but there is no relevant evidence at present.
Therefore, we explored the role of SOCCs in the pathogenesis and pathophysiological changes of OAB by studying the functional changes of SOCCs detrusor in SD rats at DO.
Method錛

本文編號(hào)：2013433