臺灣博碩士論文加值系統

摘 要
1. 在本實驗中，主要探討不同種類已上市的降低眼壓藥物，例如timolol、betaxolol、levobunolol、dipivefrin、brimonidine、latanoprost、carteolol、pilocarpine、dorzolamide、brinzolamide及unoprostone在高眼壓動物模式中，對於H2O2引起視網膜及脈絡膜中自由基的產生、視網膜中iNOS和脈絡膜中eNOS的增加及對KCl引起血管平滑肌細胞內鈣離子增加所產生的影響。
2. 實驗中所測試的降眼壓藥濃度是以藥水原濃度，經稀釋成1/100, 1/1000及1/10000。
3. 兔子眼睛經100 mmHg且維持3小時的升壓狀態後，視網膜及脈絡膜中nitric oxide synthase（NOS）及自由基產生量皆顯著的提升。
4. 利用western blotting分析視網膜神經節細胞之表面抗原，Thy-1，結果顯示Thy-1 protein在經過3小時的高眼壓狀態下，於視網膜的表現並未有顯著的差異。
5. 在正常眼壓視網膜中，timolol不論濃度的高低都有抑制H2O2引起自由基產生的效果，1/100及1/1000稀釋液之carteolol、dipivefrin及brinzolamide也可抑制正常眼壓視網膜中自由基的產生，而1/100稀釋液之brimonidine、dorzolamide及levobunolol只有在最高濃度下才有抑制效果。但1/100及1/1000稀釋液之unoprostone卻增加自由基的產生。
6. 在高眼壓視網膜中，dipivefrin及brinzolamide不論濃度的高低都有抑制H2O2引起自由基產生的效果，1/100及1/1000稀釋液之timolol及levobunolol對自由基的產生也有抑制作用，而1/100稀釋液之brimonidine、dorzolamide、betaxolol及carteolol只在最高濃度下才有抑制效果。但unoprostone卻增加自由基的產生。
7. 在正常眼壓脈絡膜中，dipivefrin及brinzolamide三種稀釋濃度都可抑制自由基的產生，1/100及1/1000之betaxolol及levobunolol也具抑制作用，而carteolol、timolol及dorzolamide只有在1/100的稀釋濃度下才有抑制效果，但latanoprost卻增加自由基的產生。
8. 在高眼壓脈絡膜中，dipivefrin、levobunolol及dorzolamide三種稀釋濃度都能抑制自由基的產生。1/100及1/1000稀釋液之carteolol、brinzolamide及unoprostone也能抑制自由基的產生，而1/100稀釋液之brimonidine、betaxolol及timolol只有在最高濃度下才有抑制效果，但高濃度latanoprost卻增加自由基的產生。
9. pilocarpine及benzalkonium chloride對於H2O2引起正常眼壓、高眼壓視網膜及脈絡膜中自由基的產生則完全無抑制效果。
10. 在高眼壓動物模式中發現，1/100及1/1000稀釋液之carteolol、timolol及pilocarpine可抑制高眼壓視網膜中iNOS的增加。
11. 在高眼壓脈絡膜中，1/100稀釋液之dipivefrin可抑制eNOS的活性，而1/100及1/1000稀釋液之pilocarpine也具抑制效果。此外，結果也顯示1/100及1/1000稀釋液之betaxolol、brinzolamide及dorzolamide可增加高眼壓脈絡膜中eNOS的活性。
12. 結果顯示，brimonidine、betaxolol、carteolol、timolol、levobunolol及pilocarpine不論濃度的高低都有抑制KCl引起細胞外鈣離子流入的效果。此外，1/100及1/1000稀釋液之brinzolamide也具抑制作用，而1/100稀釋液之dorzolamide及unoprostone只有在最高濃度下表現出抑制效果。dipivefrin、latanoprost和benzalkonium chloride對於KCl引起細胞外鈣離子流入的作用則完全無抑制效果。
13. 綜合以上結果顯示，timolol可避免視網膜中產生過多的自由基及iNOS，而betaxolol可抑制血管平滑肌收縮及增加eNOS的活性，因而具有神經保護的特性，因為這兩種藥物藥物兼具降眼壓及神經保護的作用，因此用於青光眼疾病之治療為較理想的選擇。

ABSTRACT
1. In the present study, we investigated the effects of commercial antiglaucoma drugs including timolol, betaxolol, levobunolol, dipivefrin, brimonidine, latanoprost, carteolol, pilocarpine, dorzolamide, brinzolamide and unoprostone on free radical production, iNOS and eNOS activity in retina and choroid with ocular hypertension rabbits. The effects of drugs on KCl-induced [Ca2+]i increase in vascular smooth muscle cells were also investigated with A7r5 cells.
2. All drugs have been diluted to 1/100, 1/000 and 1/10000 from original concentrations.
3. The great amount of free radical and the nitric oxide synthase (NOS) were increased after intraocular pressure (IOP) was increased to 100 mmHg for 3 hours.
4. By western blotting assay, the Thy-1 protein was not changed significantly after IOP was increased to 100 mmHg for 3 hours.
5. In retina with normal IOP, three diluted drugs of timolol and diluted to 1/100 and 1/1000 of carteolol, dipivefrin and brinzolamide were potently inhibited H2O2-induced free radical production. Only 1/100 diluted of brimonidine, dorzolamide and levobunolol inhibited the free radical production. However, unoprostone increased the free radical production.
6. In retina with high IOP, dipivefrin and brinzolamide (1/100 to 1/10000) were inhibited free radical production. In the presence of timolol and levobunolol (1/100 and 1/1000), the free radical was inhibited significantly. Only 1/100 of brimonidine, dorzolamide, carteolol and betaxolol inhibited the free radical production in high IOP retina.
7. In choroid with normal IOP, dipivefrin and brinzolamide (1/100 to 1/10000) were potently inhibited free radical production. Betaxolol and levobunolol (1/100 and 1/1000) was potently inhibited free radical production. Only 1/100 of carteolol, timolol and dorzolamide inhibited the free radical production in normal IOP choroid. However, the free radical was increased by latanoprost.
8. In choroid with high IOP, dipivefrin, levobunolol and dorzolamide (1/100 to 1/10000) were potently inhibited free radical production. In the presence of carteolol, brinzolamide and unoprostone (1/100 and 1/1000), the free radical was inhibited significantly. Only 1/100 of brimonidine, betaxolol and timolol inhibited the free radical production. However, the free radical was increased by latanoprost.
9. Pilocarpine and benzalkonium chloride have no effects to H2O2-induced free radical production in retina and choroid.
10. In rabbits with high IOP, carteolol, timolol and pilocarpine (1/100 and 1/1000) were potently inhibited the high IOP-induced iNOS production.
11. The eNOS in high IOP choroid was inhibited by 1/100 of dipivefrin and pilocarpine (1/100 and 1/1000). Betaxolol, brinzolamide and dorzolamide (1/100 and 1/1000) increased eNOS activity in high IOP choroid.
12. It was found that 50 mM KCl-induced [Ca2+]i increased in A7r5 aortic smooth muscle cells was potently inhibited by three diluted concentrations of brimonidine, betaxolol, carteolol, timolol, levobunolol and pilocarpine. Brinzolamide (1/100 and 1/1000) and 1/100 of dorzolamide and unoprostone also have the effect to inhibit the KCl-induced [Ca2+]i increase. However, dipivefrin, latanoprost and benzalkonium chloride did not inhibit the KCl-induced [Ca2+]i increase in A7r5 cells.
13. In summary, timolol possesses the effects to inhibit the iNOS and free radical production in retina. Betaxolol possesses the effects to inhibit the vasoconstriction in vessels and increase eNOS activity in choroid. Both drugs may be considered firstly in the treatment of glaucoma patients with ocular antihypertensive and neuroprotective effects simultaneously.

目錄
1. 摘要-----------------------------------------------------1
2. 英文摘要-----------------------------------------------3
3. 緒論-----------------------------------------------------5
4. 實驗材料---------------------------------------------12
5. 實驗方法---------------------------------------------15
6. 實驗結果---------------------------------------------26
7. 實驗結果總整理------------------------------------38
8. 討論---------------------------------------------------41
9. 結論---------------------------------------------------49
10. 附圖---------------------------------------------------50
11. 參考文獻---------------------------------------------90

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