间质性膀胱炎动物模型去势对其膀胱肥大细胞的影响研究

发布时间：2018-06-15 23:54

  本文选题：膀胱炎 + 间质性　； 参考：《天津医科大学》2012年硕士论文

【摘要】：目的： 1、通过膀胱内灌注硫酸鱼精蛋白(Protamine sulfate, PS)建立间质性膀胱炎(Interstitial Cystitis, IC)动物模型。 2、通过研究双侧卵巢切除对IC大鼠模型膀胱形态学、膀胱肥大细胞数目及状态的变化,来明确女性激素在间质性膀胱炎发病中的作用。 材料及方法： 1.实验对象 成年雌性SD大鼠,鼠龄6-8周,体重193～255克,平均233.5g。 2.主要试剂 硫酸鱼精蛋白粉剂(Protamine sulfate Grade X):SIGMA公司；血清17β-雌二醇Elisa试剂盒：IBL公司；血清孕激素Elisa试剂盒：IBL公司；甲苯胺蓝：天津市泌尿外科研究所提供。 3.方法 30只成年雌性SD大鼠随机分为3组：卵巢切除+硫酸鱼精蛋白组(OVX+PS组)、假手术+PS组(SHAM+PS组)和正常对照组(假手术+生理盐水组,SHAM+NS组)。三组大鼠手术后间断测定血清17β-雌二醇与孕激素水平,当雌性激素水平恒定后(约一个月)分别给予硫酸鱼精蛋白或生理盐水膀胱灌注。60天后处死大鼠并切取膀胱,HE染色观察膀胱粘膜变化及炎性细胞浸润情况,甲苯胺蓝染色标记并计数肥大细胞,透射电镜观察肥大细胞活性状态。 4.统计与分析 采用IBM SPSS19.0统计软件对所得数据进行统计学分析,符合正态分布的资料应用x±s描述。两组独立样本采用Student't检验,组间比较采用单因素方差分析,α=0.05。 结果： 1. OVX+PS组大鼠双侧卵巢切除后,其血清17β-雌二醇与孕酮水平均降低,约1月左右降至最低水平并维持恒定。与术前及假手术组(SHAM+PS组和SHAM+NS组)相比,差异均有统计学意义(P0.01) 2.正常对照组(SHAM+NS组)膀胱粘膜未见明显损伤,粘膜固有层未见明显炎性细胞浸润。PS灌注组(OVX+PS组与SHAM+PS组)之间膀胱粘膜改变没有明显差异,均表现出明显损伤,但损伤表现不均匀,部分区域膀胱粘膜上皮剥脱或局部溃疡形成,部分区域粘膜上皮细胞增生明显,粘膜下层可见炎性细胞浸润。 3.与正常对照组相比,OVX+PS组、SHAM+PS组膀胱粘膜固有层、逼尿肌和浆膜层肥大细胞计数均明显增多,差异均有统计学意义(P0.01)；而OVX+PS组和SHAM+PS组相比,其粘膜固有层、逼尿肌及浆膜层肥大细胞计数差异均不具有统计学意义(P0.05) 4.在粘膜固有层与逼尿肌,OVX+PS组与SHAM+NS组脱颗粒肥大细胞所占百分比均明显低于SHAM+PS组(P0.01)。在粘膜层,OVX+PS组脱颗粒肥大细胞百分比均较SHAM+NS组明显减少(P0.01)；而在逼尿肌,OVX+PS组与SHAM+NS组脱颗粒肥大细胞百分比没有明显差距,差异不具有统计学意义(P0.05)；在浆膜层,SHAM+NS组肥大细胞极少,未见明显脱颗粒肥大细胞,OVX+PS组脱颗粒肥大细胞百分比较SHAM+PS组明显下降(P0.01)。 5.透射电镜观察发现SHAM+NS组与OVX+PS组膀胱组织内绝大多数肥大细胞处于未脱颗粒状态,胞浆内含有大量均一的、完整的、电子致密的颗粒,脱颗粒肥大细胞占肥大细胞总数的30%以下。而SHAM+PS组大鼠膀胱组织内多数肥大细胞呈部分脱颗粒或完全脱颗粒状态,脱颗粒细胞可达70%以上 结论： 1、硫酸鱼精蛋白(PS)膀胱灌注可以建立病理组织学表现较为典型的IC动物模型。通过反复、长时间PS灌注,大鼠膀胱上皮细胞结构形态发生持续性改变,膀胱粘膜固有层、逼尿肌甚至浆膜层均可见肥大细胞增多与脱颗粒。上述改变与在IC患者膀胱变化相似。这一膀胱上皮损伤模型的建立为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础。 2、女性激素(雌激素和孕激素)对IC患者膀胱粘膜的损伤和膀胱壁各层组织肥大细胞数量的变化没有明显的影响,但是可显著影响IC患者膀胱内肥大细胞的活化。因此降低IC患者体内女性激素的含量或拮抗女性激素的作用可能会对IC患者症状的减轻起到积极作用。
[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锛，

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