本文選題：膀胱炎 + 間質(zhì)性��； 參考：《天津醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的： 1、通過膀胱內(nèi)灌注硫酸魚精蛋白(Protamine sulfate, PS)建立間質(zhì)性膀胱炎(Interstitial Cystitis, IC)動(dòng)物模型。 2、通過研究雙側(cè)卵巢切除對(duì)IC大鼠模型膀胱形態(tài)學(xué)、膀胱肥大細(xì)胞數(shù)目及狀態(tài)的變化,來明確女性激素在間質(zhì)性膀胱炎發(fā)病中的作用。 材料及方法： 1.實(shí)驗(yàn)對(duì)象 成年雌性SD大鼠,鼠齡6-8周,體重193～255克,平均233.5g。 2.主要試劑 硫酸魚精蛋白粉劑(Protamine sulfate Grade X):SIGMA公司；血清17β-雌二醇Elisa試劑盒：IBL公司；血清孕激素Elisa試劑盒：IBL公司；甲苯胺藍(lán)：天津市泌尿外科研究所提供。 3.方法 30只成年雌性SD大鼠隨機(jī)分為3組：卵巢切除+硫酸魚精蛋白組(OVX+PS組)、假手術(shù)+PS組(SHAM+PS組)和正常對(duì)照組(假手術(shù)+生理鹽水組,SHAM+NS組)。三組大鼠手術(shù)后間斷測定血清17β-雌二醇與孕激素水平,當(dāng)雌性激素水平恒定后(約一個(gè)月)分別給予硫酸魚精蛋白或生理鹽水膀胱灌注。60天后處死大鼠并切取膀胱,HE染色觀察膀胱粘膜變化及炎性細(xì)胞浸潤情況,甲苯胺藍(lán)染色標(biāo)記并計(jì)數(shù)肥大細(xì)胞,透射電鏡觀察肥大細(xì)胞活性狀態(tài)。 4.統(tǒng)計(jì)與分析 采用IBM SPSS19.0統(tǒng)計(jì)軟件對(duì)所得數(shù)據(jù)進(jìn)行統(tǒng)計(jì)學(xué)分析,符合正態(tài)分布的資料應(yīng)用x±s描述。兩組獨(dú)立樣本采用Student't檢驗(yàn),組間比較采用單因素方差分析,α=0.05。 結(jié)果： 1. OVX+PS組大鼠雙側(cè)卵巢切除后,其血清17β-雌二醇與孕酮水平均降低,約1月左右降至最低水平并維持恒定。與術(shù)前及假手術(shù)組(SHAM+PS組和SHAM+NS組)相比,差異均有統(tǒng)計(jì)學(xué)意義(P0.01) 2.正常對(duì)照組(SHAM+NS組)膀胱粘膜未見明顯損傷,粘膜固有層未見明顯炎性細(xì)胞浸潤。PS灌注組(OVX+PS組與SHAM+PS組)之間膀胱粘膜改變沒有明顯差異,均表現(xiàn)出明顯損傷,但損傷表現(xiàn)不均勻,部分區(qū)域膀胱粘膜上皮剝脫或局部潰瘍形成,部分區(qū)域粘膜上皮細(xì)胞增生明顯,粘膜下層可見炎性細(xì)胞浸潤。 3.與正常對(duì)照組相比,OVX+PS組、SHAM+PS組膀胱粘膜固有層、逼尿肌和漿膜層肥大細(xì)胞計(jì)數(shù)均明顯增多,差異均有統(tǒng)計(jì)學(xué)意義(P0.01)；而OVX+PS組和SHAM+PS組相比,其粘膜固有層、逼尿肌及漿膜層肥大細(xì)胞計(jì)數(shù)差異均不具有統(tǒng)計(jì)學(xué)意義(P0.05) 4.在粘膜固有層與逼尿肌,OVX+PS組與SHAM+NS組脫顆粒肥大細(xì)胞所占百分比均明顯低于SHAM+PS組(P0.01)。在粘膜層,OVX+PS組脫顆粒肥大細(xì)胞百分比均較SHAM+NS組明顯減少(P0.01)；而在逼尿肌,OVX+PS組與SHAM+NS組脫顆粒肥大細(xì)胞百分比沒有明顯差距,差異不具有統(tǒng)計(jì)學(xué)意義(P0.05)；在漿膜層,SHAM+NS組肥大細(xì)胞極少,未見明顯脫顆粒肥大細(xì)胞,OVX+PS組脫顆粒肥大細(xì)胞百分比較SHAM+PS組明顯下降(P0.01)。 5.透射電鏡觀察發(fā)現(xiàn)SHAM+NS組與OVX+PS組膀胱組織內(nèi)絕大多數(shù)肥大細(xì)胞處于未脫顆粒狀態(tài),胞漿內(nèi)含有大量均一的、完整的、電子致密的顆粒,脫顆粒肥大細(xì)胞占肥大細(xì)胞總數(shù)的30%以下。而SHAM+PS組大鼠膀胱組織內(nèi)多數(shù)肥大細(xì)胞呈部分脫顆粒或完全脫顆粒狀態(tài),脫顆粒細(xì)胞可達(dá)70%以上 結(jié)論： 1、硫酸魚精蛋白(PS)膀胱灌注可以建立病理組織學(xué)表現(xiàn)較為典型的IC動(dòng)物模型。通過反復(fù)、長時(shí)間PS灌注,大鼠膀胱上皮細(xì)胞結(jié)構(gòu)形態(tài)發(fā)生持續(xù)性改變,膀胱粘膜固有層、逼尿肌甚至漿膜層均可見肥大細(xì)胞增多與脫顆粒。上述改變與在IC患者膀胱變化相似。這一膀胱上皮損傷模型的建立為IC以及腺性膀胱炎、放射性膀胱炎等其他膀胱炎性疾病的研究提供了成熟的模型基礎(chǔ)。 2、女性激素(雌激素和孕激素)對(duì)IC患者膀胱粘膜的損傷和膀胱壁各層組織肥大細(xì)胞數(shù)量的變化沒有明顯的影響,但是可顯著影響IC患者膀胱內(nèi)肥大細(xì)胞的活化。因此降低IC患者體內(nèi)女性激素的含量或拮抗女性激素的作用可能會(huì)對(duì)IC患者癥狀的減輕起到積極作用。
[Abstract]:Objective:
1, establish a Interstitial Cystitis (IC) animal model by intravesical instillation of Protamine sulfate (PS).
2, the role of female hormones in the pathogenesis of interstitial cystitis was determined by studying the changes in the morphology of the bladder and the number and state of the bladder mast cells in the IC rat model.
Materials and methods:
1. experimental objects
Adult female SD rats aged 6-8 weeks, weighing 193~255 grams, with an average of 233.5g.
2. main reagents
Protamine sulfate Grade X (Grade X): SIGMA company; serum 17 beta estradiol Elisa Kit: IBL company; serum progesterone Elisa Kit: IBL company; toluidine blue: Tianjin Department of Urology Research Institute.
3. method
30 adult female SD rats were randomly divided into 3 groups: ovariectomy + sulfate protamine group (group OVX+PS), sham operation +PS group (group SHAM+PS) and normal control group (sham operation + physiological saline group, SHAM+NS group). The level of serum 17 beta estradiol and progesterone was measured intermittently in the three groups after operation, when the estrogen level was constant (about one month), respectively. The rats were sacrificed and the bladder was sacrificed after intravesical instillation of protamine sulfate or physiological saline.60. The changes of bladder mucosa and infiltration of inflammatory cells were observed by HE staining. Mast cells were labeled with toluidine blue and the mast cell activity was observed by transmission electron microscope.
4. statistics and analysis
IBM SPSS19.0 statistical software was used to make statistical analysis of the data. The data in accordance with the normal distribution were described by X + s. The two groups of independent samples were tested by Student't, and the single factor analysis of variance, alpha =0.05. was used.
Result錛，

本文編號(hào)：2024227