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題 名 | Calcium-Induced Changes on Crystallins in Organcultured Porcine Lens=鈣離子對組織培養的豬水晶體內晶狀體素的改變 |
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作 者 | 吳國揚; 王惠珠; 洪秀貞; | 書刊名 | The Kaohsiung Journal of Medical Sciences |
卷 期 | 14:9 1998.09[民87.09] |
頁 次 | 頁569-576 |
分類號 | 418.1 |
關鍵詞 | 組織培養; 水晶體; 眼內手術; 洗滌劑; Calcium; Crystallin protein; Lens culture; |
語 文 | 英文(English) |
中文摘要 | 在本實驗中,完整的豬水晶體在體外培養七天,並以已經商業化,經常在眼內手 術時當作洗滌用的緩衝液(BSS, balanced salt solution)當作水晶體的培養液,將水晶體養 在不同溫度及CO2濃度的培養箱內,經過七天的培養後不同的水晶體混濁度以光學密度掃描 儀,TSK HM-55樹脂及SDS-PAGE測試不同培養的條件對水晶體造成的影響,若將水晶體培養 在含Ca+2-free BSS培養液中,並維持在37℃或25℃以及含5% CO2或不含CO2的培養箱內, 或改用BSS培養液並在相同條件下,結果發現最不會引起水晶體混濁的培養條件是,使用不含 鈣離子的BSS培養液,而且將水晶體在25℃及不含CO2的培養箱中效果最好,經過七天培養 後,將水晶體內的可溶性蛋白質利用TSK HM-55樹脂將不同成分的晶狀體素分開,比較使用 BSS及Ca+2-free BSS二種培養液的晶狀體素時發現,在BSS培養下的水晶體,高分子量(HMW) 蛋白質及α-晶狀體素的百分比會增加,當培養液內鈣離子的濃度增加(4.3 mM,20 mM,50 mM,100 mM,及200 mM)這種變化更明顯,關於β-晶狀體素的改變則是利用SDS-PAGE可以觀 察其電泳後βH蛋白質的變化,若與培養在4.3 mM及20 mM鈣離子培養液中的βH晶狀體素 做比較,經100mM鈣離子培養的βH,其20.5 kDa及21 kDa的蛋白質消失,在200 mM鈣離 子存在下,則消失了20.5 kDa,21 kDa,22 kDa及23 kDa,從以上結果顯示BSS這種常被用 在眼內手術時使用的洗滌劑,由於它的溶液內含有鈣離子,可能會增加水晶體混濁的危險性。 |
英文摘要 | In this study, intact porcine lenses were cultured in vitro for 7 days supplemented with commercial balanced salt solution (BSS) which is usually used as an irrigation solution during intraocular surgery, and the lenses were maintained under various culture conditions, e.g. temperature and CO2 concentration. The intact porcine lenses after 7 days culture were analyzed with optical density scanner, gel permeation chromatography on TSK HM-55 column and SDS-PAGE (polyacrylamide gel electrophoresis). It was found that lenses exhibited the least opacity when lenses were cultured with Ca+2-free BSS buffer, CO2-free incubator and maintained at a temperature of 25℃. After the lenses were cultured with Ca+2-free BSS or BSS medium, the composition of crystallins in lenses was separated with TSK HM-55 column. It was indicated that the percentage of high molecular weight (HMW) protein and (α-crystallin increased, and γ -crystallin decreased in lenses incubated with BSS medium compared with lenses incubated with Ca+2-free BSS medium. Following an increase in the concentration of calcium in the medium from 4.3mM, 20mM, 50mM, 100mM to 200mM, the opacity of the lens was measured with a densitometer. The changed percentage of various crystallins was similar to lenses with BSS media that increased in HMW protein and α-crystallin, decreasing inγ-crystallin. In the case of lens protein pattern, the crystallin washed from TSK HM-55 gel was separated with SDS-PAGE (polyacrylamide gel electrophoresis). It was indicated that some of proteins disappeared when lenses were incubated with various concentrations of calcium. The vanished pH proteins were 20.5 kDa at 50mM calcium, 20.5kDa and 21 kDa at 100mM, 20.5kDa, 21kDa, 22kDa and 23kDa at 200mM which were compared with the protein bands in the presence of 20mM calcium in BSS medium. This study indicates that the commercial balanced salt solution (BSS) which is usually used as an irrigating solution during intraocular operations may increase the risk for lens opacity because of the calcium contained in the solution. |
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