中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2011年
1期
32-37
,共6页
易少凌%施斌%李婉文%徐礼剑%赵春顺
易少凌%施斌%李婉文%徐禮劍%趙春順
역소릉%시빈%리완문%서례검%조춘순
晶状体上皮细胞%后囊膜混浊%多西他赛%盐酸表柔比星%盐酸吡柔比星%雷替曲塞
晶狀體上皮細胞%後囊膜混濁%多西他賽%鹽痠錶柔比星%鹽痠吡柔比星%雷替麯塞
정상체상피세포%후낭막혼탁%다서타새%염산표유비성%염산필유비성%뢰체곡새
Lens epithelial cells%Posterior capsule opaeifieation%Docetaxel%Epirubicin%Pirarubicin
背景 一些已知的抑制人晶状体上皮细胞(LECs)增生的药物由于严重的不良反应限制了其临床应用,寻找高效、安全的抑制LECs增生的药物是防治晶状体后囊膜混浊(PCO)的研究热点.目的 探讨多西他赛对LECs增生的影响,并与盐酸表柔比星、盐酸吡柔比星和雷替曲塞的作用进行比较.方法 对永生系人LECs细胞株SRA01/04进行培养及传代,将不同浓度的多西他赛、盐酸表柔比星、盐酸吡柔比星和雷替曲塞加入LECs中分别作用24、48、72 h,以MTT法评价不同浓度的多西他赛对人LECs增生的抑制作用并与其他药物进行比较.用流式细胞术分析不同浓度的多西他赛作用48 h后对LECs细胞周期的影响,采用Annexin V-FITC/PI标记法和蛋白印迹分析法评估不同浓度的多西他赛作用48 h后LECs细胞bcl-2蛋白的表达和凋亡情况.结果 8～519 pmol/L多西他赛组LECs的增生率为100%,LECs形态正常,而66 nmol/L多西他赛组干预48 h和72 h后LECs的增生率分别为34.7%和27.4%,LECs形态出现异常.23.22～523.56μmol/L雷替曲塞对人LECs的增生无明显抑制作用.多西他赛作用48 h和72 h时,半数抑制量(IC50)明显低于盐酸吡柔比星和盐酸表柔比星.多西他赛作用LECs 48 h后,随着多西他赛浓度的增高,处于G2/M期的LECs百分数明显增加,各组的总体差异有统计学意义(F=2633.05,P＜0.01);8,3 nmol/L和266 nmol/L多西他赛浓度组干预48 h后,LECs的凋亡率分别为22.4%和27.9%,较细胞对照组升高,差异均有统计学意义(χ2=20.00,P＜0.01;χ2=42.68,P＜0.01).Western blot检测表明不同浓度多西他赛干预后bcl-2蛋白在LECs中的表达条带均较对照组淡,8.3 nmol/L多西他赛组bcl-2蛋白表达降低更为明显.结论 多西他赛、盐酸表柔比星和盐酸吡柔比星均可抑制人LECs的增生.而雷替曲塞对人LECs的增生无明显的抑制作用.多西他赛对LECs增生的抑制作用强于其他药物,其作用强度呈浓度和时间依赖性.
揹景 一些已知的抑製人晶狀體上皮細胞(LECs)增生的藥物由于嚴重的不良反應限製瞭其臨床應用,尋找高效、安全的抑製LECs增生的藥物是防治晶狀體後囊膜混濁(PCO)的研究熱點.目的 探討多西他賽對LECs增生的影響,併與鹽痠錶柔比星、鹽痠吡柔比星和雷替麯塞的作用進行比較.方法 對永生繫人LECs細胞株SRA01/04進行培養及傳代,將不同濃度的多西他賽、鹽痠錶柔比星、鹽痠吡柔比星和雷替麯塞加入LECs中分彆作用24、48、72 h,以MTT法評價不同濃度的多西他賽對人LECs增生的抑製作用併與其他藥物進行比較.用流式細胞術分析不同濃度的多西他賽作用48 h後對LECs細胞週期的影響,採用Annexin V-FITC/PI標記法和蛋白印跡分析法評估不同濃度的多西他賽作用48 h後LECs細胞bcl-2蛋白的錶達和凋亡情況.結果 8～519 pmol/L多西他賽組LECs的增生率為100%,LECs形態正常,而66 nmol/L多西他賽組榦預48 h和72 h後LECs的增生率分彆為34.7%和27.4%,LECs形態齣現異常.23.22～523.56μmol/L雷替麯塞對人LECs的增生無明顯抑製作用.多西他賽作用48 h和72 h時,半數抑製量(IC50)明顯低于鹽痠吡柔比星和鹽痠錶柔比星.多西他賽作用LECs 48 h後,隨著多西他賽濃度的增高,處于G2/M期的LECs百分數明顯增加,各組的總體差異有統計學意義(F=2633.05,P＜0.01);8,3 nmol/L和266 nmol/L多西他賽濃度組榦預48 h後,LECs的凋亡率分彆為22.4%和27.9%,較細胞對照組升高,差異均有統計學意義(χ2=20.00,P＜0.01;χ2=42.68,P＜0.01).Western blot檢測錶明不同濃度多西他賽榦預後bcl-2蛋白在LECs中的錶達條帶均較對照組淡,8.3 nmol/L多西他賽組bcl-2蛋白錶達降低更為明顯.結論 多西他賽、鹽痠錶柔比星和鹽痠吡柔比星均可抑製人LECs的增生.而雷替麯塞對人LECs的增生無明顯的抑製作用.多西他賽對LECs增生的抑製作用彊于其他藥物,其作用彊度呈濃度和時間依賴性.
배경 일사이지적억제인정상체상피세포(LECs)증생적약물유우엄중적불량반응한제료기림상응용,심조고효、안전적억제LECs증생적약물시방치정상체후낭막혼탁(PCO)적연구열점.목적 탐토다서타새대LECs증생적영향,병여염산표유비성、염산필유비성화뢰체곡새적작용진행비교.방법 대영생계인LECs세포주SRA01/04진행배양급전대,장불동농도적다서타새、염산표유비성、염산필유비성화뢰체곡새가입LECs중분별작용24、48、72 h,이MTT법평개불동농도적다서타새대인LECs증생적억제작용병여기타약물진행비교.용류식세포술분석불동농도적다서타새작용48 h후대LECs세포주기적영향,채용Annexin V-FITC/PI표기법화단백인적분석법평고불동농도적다서타새작용48 h후LECs세포bcl-2단백적표체화조망정황.결과 8～519 pmol/L다서타새조LECs적증생솔위100%,LECs형태정상,이66 nmol/L다서타새조간예48 h화72 h후LECs적증생솔분별위34.7%화27.4%,LECs형태출현이상.23.22～523.56μmol/L뢰체곡새대인LECs적증생무명현억제작용.다서타새작용48 h화72 h시,반수억제량(IC50)명현저우염산필유비성화염산표유비성.다서타새작용LECs 48 h후,수착다서타새농도적증고,처우G2/M기적LECs백분수명현증가,각조적총체차이유통계학의의(F=2633.05,P＜0.01);8,3 nmol/L화266 nmol/L다서타새농도조간예48 h후,LECs적조망솔분별위22.4%화27.9%,교세포대조조승고,차이균유통계학의의(χ2=20.00,P＜0.01;χ2=42.68,P＜0.01).Western blot검측표명불동농도다서타새간예후bcl-2단백재LECs중적표체조대균교대조조담,8.3 nmol/L다서타새조bcl-2단백표체강저경위명현.결론 다서타새、염산표유비성화염산필유비성균가억제인LECs적증생.이뢰체곡새대인LECs적증생무명현적억제작용.다서타새대LECs증생적억제작용강우기타약물,기작용강도정농도화시간의뢰성.
Background Some drugs with inhibitory effect on the proliferation of lens epithelial cells have a limiting application in clinic because of their adverse response.To screen the effective and less side-effect drug for supressing LECs growth is very inportant for the prevention and treatment of after cataract.Objective This study was to explore the effects of docetaxel on LECs growth and compare its role with epirubicin hydrochloride,pirarubicin hydrochloTide and rahitrexed.Methotis Immortalized human LECs line (SRA01/04) were cultured and passaged.Different concentrations of docetaxel,epirubicin hydrochloride,pirarubicin hydrochloride and rahitrexed were added into the medium respectively for 24.48 and 72 hours.The proliferation of LECs was detect by M1Yr.Flow cytometry analysis Was used to analyze the influence of different concentrations of docetaxel on cellular cycle at 48 hours after addition of docetaxel,and Annexin V-FITC/PI marking method was used to assesse the apoptosis of LECs under the action of docetaxel.Expression of bcl-2 protein in LECs Was evaluated by Westeru blot. Result The growth rate of LECs Wag 100%in 8-519 pmol/L doeetaxel groups with the normal cell shape.Majority of abnormal cells and low growth rate were found in 66 nmoVL docetaxel group at 48 and 72 hours.The IC50 of docetaxel was lowest in 48 and 72 hours in docetaxel group in comparison to epirubicin hydrochloride and pirarubicin hydrochloride. However,no evident inhibition on LECs growth in 23.22-523.56 μmol/L of raltitrexed.At 48 hours,the percentage of LECs in G2/M phase increased as the asccnte of concentration of docetaxel,showing a significant difference among 4 groups(F=2633.05,P＜0.01).The percentage of early apoptotic cells increased to 22.4%(χ2=20.00,P＜0.01) and 27.9%(χ2=42.68,P＜0.01)from normal control 3.1% at 48 hours after LECs exposed to 8.3 nmol/L and 266 nmol/L docetaxe.The expression of bcl-2 protein in LECs was obviously weakened after addition of docetaxel,especially 8.3 nmol/L docetaxel group. Conclusion Docetaxel,epirubicin hydrochloride and pirarubicin hydrochloride can inhibit the proliferation of human LECs in vitro.But there is no supression on LECs growth inraltitrexed.Docetaxel is proved to have a strongly arrested effect on the proliferation of LECs in comparison with epirubicin hydrochloride and pirarubicin hydrochloride and play its role at concentration-and time-dependent manner.
