| 摘要: | 全球有65億的癲癇病人，每天的抗癲癇藥用量約有1014噸，雖然有研究報告指出抗癲癇藥物可能會對患者的聽力產生傷害，然而還缺乏直接的證據。在本篇論文我們以兩種臨床常用的抗癲癇藥物進行實驗，分別為carbamazepine (CBZ)、clobazam (CLO)，CBZ主要是以阻斷鈉離子通道來阻止腦神經的異常放電，CLO則是以增加抑制性神經傳導物質-GABA分泌，來抑制腦神經的異常放電。我們以斑馬魚為動物模式來觀察抗癲癇藥對於聽覺毛細胞的影響。哺乳類動物的聽覺毛細胞位於顳骨內的內耳，難以觀察，然而斑馬魚除了具有內耳毛細胞外，體表上也有側線毛細胞，具有容易觀察的優勢。我們以PTZ引發斑馬魚癲癇模式，檢驗CBZ、CLO是否抑制斑馬魚癲癇症狀的產生。另一方面也確認pentylenetetrazole (PTZ)對於斑馬魚的毛細胞是沒有毒性的，結果顯示100 μM以及200 μM CBZ和50 μM以及100 μM CLO可以明顯的延長斑馬魚胚胎進入癲癇時期的時間，也減少進入癲癇時期的斑馬魚隻數。接著我們將斑馬魚胚胎暴露在CBZ、CLO之水溶液中長期處理96 h (0-96 hpf)，或是短期處理4 h (96 hpf-100 hpf)、6 h (96 hpf-102 hpf)、24 h (72 hpf-96 hpf)、48 h (48 hpf-96 hpf)，在藥物處理過後以活體染劑FM1-43標定側線毛細胞，觀察細胞數目的變化。結果顯示斑馬魚的胚胎曝露在CBZ 100 μM以及200 μM之下96 h或是短時間4、6、24 h之後可以發現側線毛細胞有顯著減少。斑馬魚的胚胎曝露在CLO 10 μM以及100 μM之下96 h或是短時間4、6、24 h、48 h之後可以發現側線毛細胞有顯著減少。我們的研究結果顯示，CBZ、CLO可抑制斑馬魚的癲癇症狀，也會造成毛細胞死亡。
In global, there are 6.5 billion patients with epilepsy, and the daily antiepileptic usage is about 1014 tons. There are reports pointed out that antiepileptic drugs (AED) would impact patient’s hearing, but the direct evidence is lacking. In our study, we used zebrafish as our animal models to examine if AED (carbamazepine (CBZ) and clobazam (CLO)) could impair lateral line hair cells, which is expressed on the skin of zebrafish and easy to observe.
Zebrafish embryos were exposed to CBZ and CLO for 96 h (0-96 hpf), 4 h (96 hpf-100 hpf), 6 h (96 hpf-102 hpf), 24 h (72 hpf-96 hpf), 48 h (48 hpf-96 hpf) and stained with FM1-43 to label hair cells. The results showed that the number of hair cells was decreased after the exposure of 100 μM and 200 μM CBZ for 96 h, 4 h, 6 h, 24 h. In addition, the number of hair cells was decreased after the exposure of 10 μM and 100 μM CLO for 96 h, 4 h, 6 h, 24 h, 48 h.
Thereafter, we examined if CBZ and CLO relieve pentylenetetrazole (PTZ) induced zebrafish seizure model. The results showed that 100 μM and 200 μM CBZ as well as 50 μM and 100 μM CLO increased the latency entered seizure stage. Our results indicated that CBA and CLO decreased the number of hair cells and inhibit the seizure in zebrafish model. |
