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卵巢切除大鼠毒蕈碱受体和连接蛋白43表达变化作为膀胱过度活动症的机制

Changes of muscarinic receptors and connexin-43 expression as a mechanism of overactive bladder in ovariectomized rats.

作者信息

Lee Keon-Cheol

机构信息

Department of Urology, Inje University School of Medicine, Ilsanpaik Hospital, Goyang, Korea.

出版信息

World J Urol. 2015 Nov;33(11):1875-9. doi: 10.1007/s00345-015-1496-5. Epub 2015 Feb 3.

Abstract

PURPOSE

After menopause, the bladder is known to become overactive. To investigate the mechanisms involved in these changes, we examined the muscarinic receptors M2, M3 and gap junction protein connexin-43 in an ovariectomized rat bladder.

METHODS

Twenty 10-week-old female SD rats were used. Ten rats were ovariectomized, (Ovx group) and 10 rats received a sham operation (Con group). Four weeks after the operation, urodynamic tests were performed to verify overactive bladder, and the animals were killed. The body, bladder and uterus weights were measured. The bladder specimens were prepared for immunohistochemical staining for muscarinic receptors M2, M3 and connexin-43. Western blotting was also used for the same protein measurement (M2, M3 and connexin-43). A t test with a p value of 0.05 was considered significant, and SPSS 12.0 for Windows was used to analyze the data.

RESULTS

The mean body weight of the Ovx group (315.8 ± 18.1 g) was heavier than the Con group (270.0 ± 23.6 g) (p = 0.009). The mean uterus weight of the Ovx group (260.4 ± 186.8 g) was lighter than the Con group, (600.6 ± 175.9 g) (p = 0.028) and the mean bladder weight of the Ovx group (80.2 ± 15.9 g) was lighter than the Con group (97.4 ± 10.6 g) (p = 0.041). The mean bladder contraction of the Ovx group (5.5 ± 2.3/10 min) was more frequent than that of the Con group (3.2 ± 2.8) (p < 0.05). The expressions of M2 and M3 were not different between the Ovx and the Con group, but the expression of connexin-43 in the Ovx group was more intense than in the Con group in immunohistochemical staining. These findings were also confirmed by Western blotting results.

CONCLUSIONS

Ovariectomized rats showed frequent bladder contraction and increased connexin-43 expression without changes in M2 and M3 receptor expression. These results imply that ovariectomy-induced overactive bladder may be due to an altered gap junction protein function rather than muscarinic receptor modification.

摘要

目的

已知绝经后膀胱会变得过度活跃。为了研究这些变化所涉及的机制,我们检测了去卵巢大鼠膀胱中的毒蕈碱受体M2、M3和缝隙连接蛋白连接蛋白43。

方法

使用20只10周龄雌性SD大鼠。10只大鼠接受去卵巢手术(去卵巢组),10只大鼠接受假手术(对照组)。手术后4周,进行尿动力学测试以验证膀胱过度活动症,然后处死动物。测量体重、膀胱重量和子宫重量。制备膀胱标本用于毒蕈碱受体M2、M3和连接蛋白43的免疫组织化学染色。Western印迹法也用于相同蛋白质的测量(M2、M3和连接蛋白43)。p值为0.05的t检验被认为具有显著性,使用SPSS 12.0 for Windows软件分析数据。

结果

去卵巢组的平均体重(315.8±18.1 g)高于对照组(270.0±23.6 g)(p = 0.009)。去卵巢组的平均子宫重量(260.4±186.8 g)低于对照组(600.6±175.9 g)(p = 0.028),去卵巢组的平均膀胱重量(80.2±15.9 g)低于对照组(97.4±10.6 g)(p = 0.041)。去卵巢组的平均膀胱收缩频率(5.5±2.3/10分钟)高于对照组(3.2±2.8)(p < 0.05)。在免疫组织化学染色中,去卵巢组和对照组之间M2和M3的表达没有差异,但去卵巢组中连接蛋白43的表达比对照组更强。Western印迹结果也证实了这些发现。

结论

去卵巢大鼠表现出频繁的膀胱收缩和连接蛋白43表达增加,而M2和M3受体表达没有变化。这些结果表明,去卵巢诱导的膀胱过度活动症可能是由于缝隙连接蛋白功能改变而非毒蕈碱受体修饰所致。

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