CAJ | 학술논문

目的研究大鼠慢性眼拍打伤后晶状体可溶性蛋白与不溶性蛋白的改变,并研究大鼠热休克或喂饲HSP70阻滞剂Quercetin后对钝挫伤性晶状体蛋白的影响.方法实验研究.SpragueDawley (SD) 大鼠24只(24只眼),完全随机设计分成以下4组:A组(对照组):6只眼;B组(拍打组):6只眼,每次以20 g钢球20 cm高度拍打大鼠右眼100回,每周1次,连续5周;C组(热休克组):6只眼,温水浴(45℃)使大鼠体温提高至40.5～41.5℃ 8 min,常温下恢复2～3 h后拍打眼球同上.每周重复1次,连续5周;D组(Quercetin组):6只眼,喂饲大鼠Quercetin 100 mg/kg体重,2～3 h后拍打眼球同上.每周重复1次,连续5周.蛋白定量用Bradford法.对晶状体蛋白测定结果 ,采用完全随机设计的方差分析,并用q检验方法进行组间两两比较.对透明晶状体和混浊晶状体蛋白测定结果采用成组设计定量资料t检验进行分析.结果拍打眼球5周后,热休克组的可溶性晶状体蛋白含量为22.71±1.99,较其他3组明显升高,差异有统计学意义(F=37.82,P＜0.01);拍打组的不可溶性蛋白含量为2.60±0.48,较其他3组高,差异有统计学意义(F=3.86,P＜0.05).可溶性晶状体蛋白的十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析发现混浊晶状体眼在66 000处出现明显的高分子量蛋白带,随白内障程度增加更显著.结论慢性拍打眼球可造成晶状体损伤,不可溶性蛋白含量增加.热休克可增加品状体可溶性蛋白含量,减少不可溶性蛋白.可溶性蛋白SDSPAGE分析结果可以看出慢性拍打眼球引起晶状体蛋白质的变化向高分子蛋白移动.
목적 연구대서만성안박타상후정상체가용성단백여불용성단백적개변,병연구대서열휴극혹위사HSP70조체제Quercetin후대둔좌상성정상체단백적영향.방법 실험연구.SpragueDawley (SD) 대서24지(24지안),완전수궤설계분성이하4조:A조(대조조):6지안;B조(박타조):6지안,매차이20 g강구20 cm고도박타대서우안100회,매주1차,련속5주;C조(열휴극조):6지안,온수욕(45℃)사대서체온제고지40.5～41.5℃ 8 min,상온하회복2～3 h후박타안구동상.매주중복1차,련속5주;D조(Quercetin조):6지안,위사대서Quercetin 100 mg/kg체중,2～3 h후박타안구동상.매주중복1차,련속5주.단백정량용Bradford법.대정상체단백측정결과 ,채용완전수궤설계적방차분석,병용q검험방법 진행조간량량비교.대투명정상체화혼탁정상체단백측정결과 채용성조설계정량자료t검험진행분석.결과 박타안구5주후,열휴극조적가용성정상체단백함량위22.71±1.99,교기타3조명현승고,차이유통계학의의(F=37.82,P＜0.01);박타조적불가용성단백함량위2.60±0.48,교기타3조고,차이유통계학의의(F=3.86,P＜0.05).가용성정상체단백적십이완기류산납취병희선알응효전영(SDS-PAGE)분석발현혼탁정상체안재66 000처출현명현적고분자량단백대,수백내장정도증가경현저.결론 만성박타안구가조성정상체손상,불가용성단백함량증가.열휴극가증가품상체가용성단백함량,감소불가용성단백.가용성단백SDSPAGE분석결과 가이간출만성박타안구인기정상체단백질적변화향고분자단백이동.
Objective To study the water soluble and insoluble lens proteins in contusion cataract of rat model, and to study the influence of heat shock (thermotolerance) or quercetin (heat shock protein inhibitor) on the lens proteins.Methods Spragne-Dawley (SD) rats were randomized into four groups(6in each group):(1) A group,control group.(2) B group,contusion group.(3) C group,heat shock group.(4)D group,quercetin group.In contusion group,contusion of eyes was induced by dropping steel ball against the eyeball (20 g ball dropped from 20 cm height for 100 times each week for 5 weeks).In heat shock group, heat shock was induced by raising the body temperature to 40.5-41.5℃ for 8 min,2-3 h before the eye contusion (as previously described).Heat shock and contusion was given each week for 5 weeks.In quercetin group,rats were fed with quercetin (100 mg/kg weight) 2-3 h before the contusion.Quercetin and contusion was given each week for 5 weeks.Amount of proteins was measured by using Bradford methods.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used for the separation and analysis of lens proteins.Results Five weeks after contusion, the mean water soluble protein of heat shock group was 22.7 1±1.99, increased significantly than that of the other three groups.The mean insoluble protein of contusion group was 2.60±0.48, increased than that of the other three groups.From the analysis of SDS-PAGE,we found that there was a high molecular band near 66 KD in opaque lens and the band became more prominent with the growing opacification of the lens.Conclusions In contusion cataract,water insduble lens protein is increased and water soluble lens protein is decreased.SDS-PAGE showed that lens injury resulted in lens proteins moving toward high molecular band.