膀胱出口梗阻后大鼠逼尿肌细胞内质网形态及GRP78表达的研究

发布时间：2018-03-06 18:26

本文选题：膀胱出口部分梗阻　切入点：膀胱重构　出处：《山西医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的观察膀胱出口梗阻后膀胱重构中,逼尿肌平滑肌细胞内质网形态及内质网应激标志性蛋白——葡萄糖调节蛋白78(GRP78)的表达情况,探讨内质网应激在膀胱重构机制中可能的作用。方法雄性Wistar大鼠40只,随机分为假手术组10只,梗阻2周组15只,梗阻4周组15只,采用经会阴途径球部尿道部分结扎的方法建立膀胱出口部分梗阻模型。采用透射电子显微镜观察逼尿肌平滑肌细胞的超微结构,通过免疫组化和real-time PCR分别检测逼尿肌中GRP78蛋白及mRXA表达变化。结果假手术组、梗阻2周组和梗阻4周组大鼠膀胱逼尿肌细胞GRP78蛋白阳性表达率为(3.37±0.38)%、(51.22±0.27)%、(27.02±1.85)%,梗阻2周组大鼠膀胱逼尿肌中GRP78阳性表达明显增多,与假手术组相比有明显差异(P0.01),梗阻4周组大鼠逼尿肌中GRP78阳性表达增多,与假手术组相比有明显差异(P0.01),但比梗阻2周组表达量减少(P0.01)。各组GRP78mRNA表达量分别为(22.21±1.58)、(30.62±2.49)、(24.52±2.24),梗阻2周组大鼠膀胱逼尿肌细胞GRP78mRNA表达较假手术组明显增加(P0.01)；与梗阻2周组相比,梗阻4周组大鼠膀胱逼尿肌细胞GRP78mRNA表达量下降,有显著性统计学差异(P0.01),梗阻4周组与假手术组相比无统计学差异。结论采用会阴途径球部尿道部分结扎方法能成功建立雄性大鼠膀胱出口梗阻动物模型。在梗阻后膀胱重构过程中,逼尿肌平滑肌细胞内质网形态有明显损伤性变化,GRP78蛋白及mRNA表达水平明显增加,提示内质网应激参与膀胱重构的进展。
[Abstract]:Objective to observe the morphology of endoplasmic reticulum (ER) and the expression of endoplasmic reticulum (ER) in detrusor smooth muscle cells (SMCs) after bladder outlet obstruction, and to investigate the role of endoplasmic reticulum stress (ER) in bladder remodeling. Methods 40 male Wistar rats were randomly divided into sham operation group (n = 10), obstruction group (n = 15) and obstruction group (n = 15). The model of bladder outlet obstruction was established by transperineal partial ligation of the bulbar urethra. The ultrastructure of detrusor smooth muscle cells was observed by transmission electron microscope. The changes of GRP78 protein and mRXA expression in detrusor muscle were detected by immunohistochemistry and real-time PCR, respectively. Results. In the sham operation group, the positive expression rate of GRP78 protein in the bladder detrusor muscle cells was 3.37 卤0.38 in the obstruction 2 weeks group and 4 weeks in the obstruction group. The positive expression rate of GRP78 was significantly increased in the bladder detrusor muscle of the rats in the obstruction 2 weeks group (51.22 卤0.27) and the bladder detrusor muscle in the obstruction 2 weeks group (27.02 卤1.85). Compared with the sham operation group, there was a significant difference in the expression of GRP78 in the detrusor muscle of rats in the 4-week obstructive group, and the positive expression of GRP78 in the detrusor muscle was increased. Compared with the sham operation group, there was a significant difference between the two groups (P 0.01), but the expression of GRP78mRNA was decreased than that of the obstruction group at 2 weeks. The expression of GRP78mRNA in each group was 22.21 卤1.58, 30.62 卤2.49 and 24.52 卤2.240.The GRP78mRNA expression of bladder detrusor cells in the obstruction group was significantly higher than that in the sham operation group, which was significantly higher than that in the sham operation group, and was significantly higher than that in the obstruction group at 2 weeks. The expression of GRP78mRNA in bladder detrusor cells in the 4-week group was significantly lower than that in the sham operation group (P 0.01), but there was no significant difference between the 4 week group and the sham operation group. Conclusion. The animal model of bladder outlet obstruction in male rats can be successfully established by partial urethral ligation via the perineal approach. The changes of endoplasmic reticulum in detrusor smooth muscle cells showed that the expression of GRP78 protein and mRNA increased significantly, suggesting that endoplasmic reticulum stress was involved in bladder remodeling.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R694.2

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