磷酸二酯酶（PDEs）在大鼠膀胱中的表达与功能
Expression and function of phosphodiesterases (PDEs) in the rat urinary bladder.
作者信息
Zhu Xiaofei, Zhai Kui, Mi Yue, Ji Guangju
机构信息
Department of Urology, Beijing Jishuitan Hospital, Beijing, China.
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Beijing, 100101, China.
出版信息
BMC Urol. 2017 Jul 7;17(1):54. doi: 10.1186/s12894-017-0244-0.
BACKGROUND
It has been shown that hosphodiesterases (PDEs) play an important role in mediating the smooth muscle tone of rat urinary bladder. However, the gene expression profiles of them were still unknown.
METHODS
Urinary bladder Strips were obtained from both neonatal and adult Sprague-Dawley rats. RT-PCR/western blot and organ bath were used to measure the expression and function of PDEs.
RESULTS
Adult rat urinary bladder expressed various PDE mRNA with the following rank order: PDE5A ≈ PDE9A ≈ PDE10A > PDE2A ≈ PDE4A ≈ PDE4D > PDE4B ≈ PDE3B ≈ PDE8B ≈ PDE7A ≈ PDE7B > PDE1A. PDE1B, PDE1C, PDE3A, PDE4C, PDE8A, and PDE11A were not detected. Of interest, the mRNA and protein of PDE3A were significantly decreased in adult rat urinary bladder compared to neonatal rat urinary bladder. Cilostamide, a specific inhibitor for PDE3, significantly inhibited the amplitude and frequency of carbachol-enhanced phasic contractions of neonatal rat bladder strips by 38.8% and 12.1%, respectively. Compared to the neonatal rat bladder, the effect of cilostamide was significantly blunted in adult rat urinary bladder: the amplitude and frequency of carbachol-enhanced phasic contractions were decreased by 13.4% (P < 0.01 vs neonatal rat bladder) and 4.4%, respectively. However, the mRNA and the protein levels of PDE3B were similar between neonatal and adult rat bladder.
CONCLUSION
We found that several PDE isoforms were expressed in the rat urinary bladder with distinct levels. Moreover, we showed that the function of PDE3 was blunted in adult rat bladder likely due to the decreased expression of PDE3A.
背景
已有研究表明，磷酸二酯酶（PDEs）在介导大鼠膀胱平滑肌张力方面发挥重要作用。然而，其基因表达谱仍不清楚。
方法
从新生和成年Sprague-Dawley大鼠获取膀胱条。采用逆转录聚合酶链反应/蛋白质免疫印迹法（RT-PCR/ Western blot）和器官浴槽法测量PDEs的表达和功能。
结果
成年大鼠膀胱表达多种PDE mRNA，其表达水平排序如下：PDE5A≈PDE9A≈PDE10A＞PDE2A≈PDE4A≈PDE4D＞PDE4B≈PDE3B≈PDE8B≈PDE7A≈PDE7B＞PDE1A。未检测到PDE1B、PDE1C、PDE3A、PDE4C、PDE8A和PDE11A。有趣的是，与新生大鼠膀胱相比，成年大鼠膀胱中PDE3A的mRNA和蛋白质水平显著降低。西洛酰胺是一种PDE3特异性抑制剂，可分别显著抑制新生大鼠膀胱条中卡巴胆碱增强的相性收缩幅度和频率38.8%和12.1%。与新生大鼠膀胱相比，西洛酰胺对成年大鼠膀胱的作用明显减弱：卡巴胆碱增强的相性收缩幅度和频率分别降低13.4%（与新生大鼠膀胱相比，P＜0.01）和4.4%。然而，新生和成年大鼠膀胱中PDE3B的mRNA和蛋白质水平相似。
结论
我们发现大鼠膀胱中表达了几种不同水平的PDE亚型。此外，我们表明成年大鼠膀胱中PDE3的功能减弱可能是由于PDE3A表达降低所致。
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