Tacrolimus (FK-506)及急性單側輸尿管擴張對於麻醉老鼠在骨盆—尿道反射中glutamate依賴性的增益現象的影響。

Effects of tacrolimus and acute unilateral ureter distension on glutamate- dependent potentiation in pelvic-urethral reflex potentiation in anesthetized rats

陳國榮(Kuo-Jung Chen)

指導教授 ： 林則彬

中山醫學大學/醫學院/醫學研究所/博士(2007年)

https://doi.org/10.6834/CSMU.2007.00030

摘要

Tacrolimus對於骨盆—尿道反射增益現象的影響中文摘要
本實驗的目的在研究tacrolimus(FK506)，一種蛋白質磷酸化酶2B (protein phosphatase 2B) 抑制劑，對於在使用urethane麻醉的老鼠中作為研究時，對骨盆傳入神經纖維與尿道之間的反射可塑性(reflex plasticity)的影響。反覆刺激(Repetitive stimulation， RS， 頻率1赫茲) 會在骨盆—尿道反射(pelvic-urethral reflex ，PUR)誘發一個增益現象 (在控制組及反覆刺激組中，每次刺激所引起的動作電位數目分別為0.9 ± 0.2次及10.5 ± 1.6 次，P值小於零點零一， N值為 10)。椎管內注射tacrolimus (100微莫耳， 10tacrolimus組，每次刺激引起的動作電位數目為3.2 ± 0.9 次，相較於反覆刺激組每次刺激引起的動作電位數目為10.5 ± 1.6次，兩者相比，P值小於零點零一， N值為10) 。椎管內注射glutamate (100微莫耳， 10微升， 一次注射) 及 N-methyl-D-aspartic acid (NMDA，100微莫耳，10微升，一次注射) ，兩者皆可以逆轉由tacrolimus所施加的阻礙反覆刺激誘發的骨盆—尿道反射活動的增益現象(在glutamate組每次刺激引起的動作電位數目為 15.0 ± 1.4次，在NMDA組每次刺激引起的動作電位數目為11.4 ± 1.4次，相較於反覆刺激組在使用tacrolimus治療過後的每次刺激引起的動作電位數目3.2 ± 0.9次， P值小於零點零一，N值為 7) 。 此外，由該兩種glutamate接受體作用劑，所展現的逆轉效果並沒有統計上顯著的差異 (P值無顯著的差異，N值為 7) 。 所有的結果顯示，tacrolimus可以阻礙 glutamatergic NMDA 接受體媒介的骨盆—尿道反射活動的增益現象。 該發現對於使用tacrolimus作為免疫抑制治療的病患而言，可能是切要的病理發現。而tacrolimus是否會在這類的病人引起尿失禁，則需要更進一步的研究。
急性單側輸尿管擴張對於骨盆—尿道反射增益現象的影響中文摘要
因輸尿管內壓力(intra-ureter pressure，IUP)的急性增加，而對於骨盆—尿道反射(pelvic-urethral reflex，PUR) 增益現象 (potentiation)的影響，將在使用urethane麻醉的老鼠身上在作骨盆神經傳入性刺激後，以記錄外尿道括約肌肌電圖(external urethra sphincter electromyogrm，EUSE ) 活動作探討。當與測試刺激 (test stimulation，TS，以1/30赫茲的頻率刺激)相比較時，反覆刺激(repetitive stimulation，RS，以1赫茲頻率刺激)會在骨盆—尿道反射誘發一個脊髓反射增益現象(spinal reflex potentiation，SRP) 。在測試刺激時，每次刺激引起的動作電位數目為1.03 ± 0.12次，而在反覆刺激時，每次刺激引起的動作電位數目為16.90 ± 2.00次者相比，有顯著的差異，P值小於零點零一，N值各為7。而該反覆刺激所引起的脊髓反射增益現象可以被椎管內注射 2,3-dihydroxy-6-nitro-7- sulfamoly-benzo (F) quinoxaline [NBQX，為一種α-amino-3- hydroxy-5-methyl-4-isoxazoleproprionat (AMPA)接受體擷抗劑]及D-2-amino-5-phosphonovalerate [APV，為一種N-methyl-D- aspartate (NMDA) 接受體擷抗劑]明顯地壓制下來(其每次刺激引起的動作電位數目，分別為在椎管內注射NBQX為11.6 ± 0.71次，及在椎管內注射APV時為 1.01 ± 0.30次，與因反覆刺激所引起的動作電位數目每秒 16.90 ± 2.00次相比，有顯著的差異，P值小於零點零一，N值各為7) 。急性階梯式的增加輸尿管內壓力，會以依劑量多寡的方式(dose dependent manner)逐漸地減弱，而最後廢除反覆刺激誘發的反射增益現象(在輸尿管內壓力分別為0， 2.5， 5， 7.5， 10， 12.5，15， 17.5及 20 公分水柱時，其相對的每次刺激所誘發的動作電位數目分別為 16.80 ± 1.30 ， 17.00 ± 1.30， 16.30 ± 1.30， 10.50 ± 1.80, 8.80 ± 1.90 ， 3.50 ± 1.60 ，0.80 ± 0.20 ，0.70 ± 0.20及 0.20 ± 0.10次，在每一個輸尿管內壓力值時，N值各為7) 。椎管內注射NMDA (一種glutamate NMDA接受體作用劑) 及bicuculline [一種gamma amino-butyric acid (GABA)接受體擷抗劑]時，兩者都可以逆轉因為單側輸尿管擴張而造成對於反覆刺激所誘發的脊髓反射增益現象的抑制作用(其每次刺激引起的動作電位數目分別為椎管內注射NMDA時為14.0 ± 4.04次，及椎管內注射bicuculline時為8.00 ± 1.53次，兩者與因單側輸尿管擴張而造成的動作電位數目相比，有顯著的差異；而且兩者相比較，也有顯著的差異，P值小於零點零一，N值各為7) 。以上的結果顯示，單側輸尿管擴張可以經由GABA系統所媒介的反應抑制脊髓NMDA依賴性的骨盆—尿道反射增益現象，而代償性地放鬆尿道。

關鍵字

tacrolimus(FK506) ； 骨盆—尿道反射 ； 增益現象 ； 急性單側輸尿管擴張

並列摘要

Tacrolimus對於骨盆—尿道反射增益現象的英文摘要
Effects of tacrolimus, a protein phosphatase 2B inhibitor, on the reflex plasticity between the pelvic afferent nerve fibers and the urethra were examined in urethane anesthetized rats. Repetitive stimulation (1 Hz) induced a potentiation (0.9 ± 0.2 and 10.5 ± 1.6 spikes in control and repetitive stimulation groups, respectively, P< 0.01, N= 10) in the activities of the pelvic-urethral reflex. Intrathecal tacrolimus (100 μM, 10μl, bolus) blocked repetitive stimulation- induced potentiation in pelvic-urethral reflex activities (3.2 ± 0.9 spikes in tacrolimus group versus 10.5 ± 1.6 spikes in repetitive stimulation group, P< 0.01, N= 10). Glutamate (intrathecal, 100μM, 10μl, bolus) and N-methyl-D-aspartic acid (intrathecal, 100μM10μl, bolus) both reversed the blocking effects exerted by tacrolimus on repetitive stimulation-induced pelvic-urethral reflex potentiation (15.0 ± 1.4 spikes in glutamate group and 11.4 ± 1.4 spikes in N-methyl-D-aspartic acid group versus 3.2 ± 0.9 spikes in tacrolimus-treated repetitive stimulation group, P< 0.01, N= 7). In addition, the reversal effect elicited by these two agonists of glutamate receptors showed no statistical difference (P= NS, N= 7). All these results demonstrated that tacrolimus could block glutamatergic N-methyl-D-aspartic acid receptor-mediated potentiation in pelvic-urethral reflex activities. This finding may be pathologically relevant in patients who take tacrolimus as immunosuppressant therapy. Whether tacrolimus will induce urine incontinence in such patients or not needs further investigation.

急性單側輸尿管擴張對於骨盆—尿道反射增益現象的影響英文摘要
The effects of acute increase in intra-ureter pressure (IUP) on the pelvic-urethral reflex potentiation were examined in urethane- anesthetized rats by recording the external urethra sphincter electromyogram (EUSE) activities evoked by the pelvic afferent stimulation. When compared with a single action potential elicited by the test stimulation (TS, which is characterized by an intensity that evoked a constant reflex response without facilitation, 1/30 Hz, 1.03 ± 0.12 spikes/stimulation, N= 7), the repetitive stimulation (RS, with the identical stimulation intensity as the test stimulation, 1 Hz) significantly induced spinal reflex potentiation (SRP, 16.90 ± 2.00 spikes/stimulation, P< 0.01, N= 7). Such spinal reflex potentiation was significantly attenuated by intrathecal 2, 3 -dihydroxy-6-nitro- 7-sulfamoyl- benzo (F) quinoxaline [NBQX, a glutamatergic α- amino-3-hydroxy-5-methyl-4-isoxazoleproprionat(AMPA) receptor antagonist] and D-2-amino-5-phosphonovalerate [APV, a glutamatergic N-methyl-D-aspartate (NMDA) antagonist; the spike number per stimulation:11.6 ± 0.71 for NBQX, 1.01 ± 0.30 for APV, and 16.90 ± 2.00 for RS, respectively, P< 0.01, N= 7]. Acute stepwise elevations in the intra-ureter pressure gradually attenuated and eventually abolished the repetitive stimulation- induced spinal reflex potentiation (16.80 ± 1.30, 17.00 ± 1.30, 16.30 ± 1.30, 10.50 ± 1.80, 8.80 ± 1.90, 3.50 ± 1.60, 0.80 ± 0.20, 0.70 ± 0.20 and 0.20 ± 0.10 spikes/stimulation at intra-ureter pressure of 0, 2.5, 5, 7.5, 10, 12.5, 15, 17.5 and 20 cmH2O, respectively, N=7). Intrathecal NMDA (a glutamatergic NMDA receptor agonist) and bicuculline [a gamma amino-butyric acid (GABA) receptor antagonist] both reversed the abolition on repetitive stimulation-induced spinal reflex potentiation caused by the unilateral ureter distension (14.0 ± 4.04 and 8.00 ± 1.53 spikes/stimulation, respectively, N= 7, P< 0.01). All the results suggested unilateral ureter distension might compensatory relax the urethra via a GABAergic inhibition on NMDA-dependent spinal reflex potentiation.

並列關鍵字

Tacrolimus ； ureter distension ； glutamate ； potentiation ； pelvic-urethral reflex

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