睡眠剝奪對大鼠骨盆神經-外尿道括約肌反射塑性之影響

Effects of Sleep Deprivation on the Pelvic Nerve-to-Urethra Reflex Plasticity in Rats

簡瑋槿(Wei-Ching Chien)

指導教授 ： 林則彬 ； 陳進典

中山醫學大學/醫學院/醫學研究所/碩士(2006年)

https://doi.org/10.6834/CSMU.2006.00084

摘要

骨盆神經-外尿道括約肌反射 (pelvic nerve-urethral reflex, PUR) 會引發尿道括約肌的收縮及維持儲存尿液期間的自律性，在藥理試驗也證實在骨盆神經-外尿道括約肌反射的活性是透過麩胺酸 NMDA 接受器所調控的；此外，給予睡眠剝奪後在大鼠的海馬回會減低長期增益效應 (Long-term potentiation, LTP) 或是給予快速動眼期睡眠剝奪則有抑制 LTP 的現象；一氧化氮 (NO) 會間接調控許多生理及病理上神經退化性疾病的神經突觸塑性的活性。因此本篇實驗主要目的想探討給予睡眠剝奪後，骨盆神經-外尿道括約肌反射的活性是否會有影響，並且 NO 是否參與其調控；更進ㄧ步想探討脊髓內 nNOS 的表現量之變化情形。
本實驗使用大鼠並在椎管內注射 Glutamatergic 拮抗劑及促進劑，亦注射 Nitrinergic 拮抗劑及促進劑，並取下脊髓 (L6-S2) 組織，來探討 nNOS 蛋白質表現量之變化。實驗動物經睡眠剝奪的處理後，發現反覆性刺激引發骨盆神經─外尿道括約肌反射之活性有降低之情形。Glutamatergic 拮抗劑，可抑制反覆性刺激所引發 PUR 之活性；給予睡眠剝奪後，亦有此抑制現象產生。Glutamatergic 促進劑，可增加測試性刺激所引發 PUR 之活性；給予睡眠剝奪後，亦有此增加趨勢。Nitrinergic 拮抗劑，可抑制反覆性刺激所引發 PUR 之活性；給予睡眠剝奪後，亦可抑制此現象的產生。接著再給予 Nitrinergic 促進劑，可將此被抑制的情形恢復至原本反覆性刺激所產生的現象；給予睡眠剝奪後，亦有此回復現象的產生。
從西方點墨法與免疫組織化學法的結果可以發現，在脊髓 (L6-S2) 組織中，確實有神經性一氧化氮合成酶 (nNOS) 及磷酸化神經性一氧化氮合成酶 (p-NOS) 蛋白質的表現，並在給予電刺激下，nNOS 蛋白質表現有略高的現象，此外，給予睡眠剝奪下，nNOS 蛋白質表現量有降低之情形。
由本篇實驗結果顯示，動物給予睡眠剝奪後，會降低反覆性刺激引發 PUR 之活性，推論睡眠剝奪可能對下泌尿系統儲尿功能造成傷害及神經退化的現象。

關鍵字

膀胱 ； 睡眠剝奪 ； 一氧化氮 ； 一氧化氮合成酶塑性

並列摘要

The aim of this study was to examine the participation of nitrinergic neurotransmission in the stimulation-induced potentiation on pelvic nerve-to-urethra reflex (PUR) activities and determine the neuronal nitric oxide synthase (nNOS) reactivity of lower spinal levels (L6-S2), which mediated micturition function after total sleep deprivation (TSD).The magnitude of the repetitive stimulation (RS, 1 Hz)-induced potentiation in PUR activity decreased significantly in the TSD group when compared with the control groups (19.86±1.39 and 9.43±1.39 spikes/stimulation in control and TSD group; respectively, P < 0.01, n=7). The magnitude of the RS induced potentiation in PUR activity decreased significantly after intrathecal 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX; from 15.36±1.11 to 7.21±0.74, and from 9.44±0.44 to 5.44±0.58 spikes/stimulation in control, and TSD, respectively, P<0.01, n=7) and D-2-amino-5-phosphonoraleric acid (APV, from 17.38±1.30 to 2.94±0.34, and from 9.44±0.44 to 2.25±0.31 spikes/stimulation in control, and TSD, respectively, P < 0.01, n=7). In addition, potentiation in PUR activities was induced by intrathecal L-glutamate (from 1.50±0.22 to 26.50±0.50, and from 1.33±0.21 to 5.67±0.42 spikes/stimulation in control, and TSD, respectively, P < 0.01, n=7) and N-methyl-Daspartic acid (from 1.17±0.17 to 11.50±1.50, and from 1.33±0.21 to 3.83±0.40 spikes/stimulation in control, and TSD); Intrathecal L-NAME blocked repetitive stimulation induced potentiation in pelvic-urethral reflex activities (from 14.56±0.80 to 3.67±0.71, and from 9.33±0.33 to 3.67±0.33 spikes/stimulation in control, and TSD P<0.01, n=7). Intrathecal L-Arg reversed the blocking effects exerted by L-NAME on repetitive stimulation-induced pelvic-urethral reflex potentiation (from 3.67±0.71 to 14.17±1.19, and from 3.67±0.33 to 9.67±0.33 spikes/stimulation in control, and TSD P<0.01, n=7).
nNOS activities were found in the lower spinal levels. In addition, following TSD treatment, the intensity of nNOS reactivity were reduced when compared with control group (n=5). All the results suggest that the increased nNOS in spinal levels appears to be related with the RS-induced potentiation in PUR activity and decrease of nNOS expression may deteriorate the spinal neurotransmission and result in the TSD-induced micturition dysfunction.

並列關鍵字

bladder ； sleep deprivation ； nitric oxide ； nitric oxide synthase ； plasticity

參考文獻

Agostino PV, Ferreyra GA, Murad AD, Watanabe Y, Golombek DA (2004) Diurnal, circadian and photic regulation of calcium/calmodulin-dependent kinase II and neuronal nitric oxide synthase in the hamster suprachiasmatic nuclei. Neurochem Int 44:617-25.

Anderson KE (1993) Pharmacology of lower urinary tract smooth muscles and penile erectile tissues. Pharmacol Rev 45:253-308.

Andersson KE, Arner A (2004) Urinary bladder contraction and relaxation: physiology and pathophysiology. Physiol Rev 84:935-86.

Andersson KE, Persson K (1994) Nitric oxide synthase and nitric oxide-mediated effects in lower urinary tract smooth muscles. World J Urol 12:274-80.

Ayers NA, Kapas L, Krueger JM (1996) Circadian variation of nitric oxide synthase activity and cytosolic protein levels in rat brain. Brain Res 707:127-30.