湖南医科大学学报
湖南醫科大學學報
호남의과대학학보
BULLETIN OF HUNAN MEDICAL UNIVERSITY
2001年
1期
1-3
,共3页
肖卫民%袁开宇%袁灿%王慷慨%尢家騄%肖献忠
肖衛民%袁開宇%袁燦%王慷慨%尢傢騄%肖獻忠
초위민%원개우%원찬%왕강개%왕가록%초헌충
晶体蛋白类%热休克蛋白%心肌%细胞,培养的%移位
晶體蛋白類%熱休剋蛋白%心肌%細胞,培養的%移位
정체단백류%열휴극단백%심기%세포,배양적%이위
采用热休克对新生大鼠心肌细胞进行处理,观察αB-晶状体蛋白在细胞内的分布及其表达变化。结果:①Western-blot显示热休克后胞浆中可溶性αB-晶状体蛋白迅速移位至细胞内的不溶性结构中,2h左右基本复位;②免疫双荧光细胞化学显示热休克导致αB-晶状体蛋白在细胞浆内发生分布变化,而HSC70从胞浆向胞核移位;③免疫电镜定量分析进一步证实αB-晶状体蛋白在热休克时主要移向Z线及核膜等细胞骨架部位;④Northern-blot显示热休克诱导心肌细胞αB-晶状体蛋白mRNA的表达增加。热休克处理使αB-晶状体蛋白迅速移位至细胞骨架,表明其在心肌预适应早期相中可能发挥保护作用。而αB-晶状体蛋白的诱导表达,可能是心肌预适应延迟相保护的重要机制之一。
採用熱休剋對新生大鼠心肌細胞進行處理,觀察αB-晶狀體蛋白在細胞內的分佈及其錶達變化。結果:①Western-blot顯示熱休剋後胞漿中可溶性αB-晶狀體蛋白迅速移位至細胞內的不溶性結構中,2h左右基本複位;②免疫雙熒光細胞化學顯示熱休剋導緻αB-晶狀體蛋白在細胞漿內髮生分佈變化,而HSC70從胞漿嚮胞覈移位;③免疫電鏡定量分析進一步證實αB-晶狀體蛋白在熱休剋時主要移嚮Z線及覈膜等細胞骨架部位;④Northern-blot顯示熱休剋誘導心肌細胞αB-晶狀體蛋白mRNA的錶達增加。熱休剋處理使αB-晶狀體蛋白迅速移位至細胞骨架,錶明其在心肌預適應早期相中可能髮揮保護作用。而αB-晶狀體蛋白的誘導錶達,可能是心肌預適應延遲相保護的重要機製之一。
채용열휴극대신생대서심기세포진행처리,관찰αB-정상체단백재세포내적분포급기표체변화。결과:①Western-blot현시열휴극후포장중가용성αB-정상체단백신속이위지세포내적불용성결구중,2h좌우기본복위;②면역쌍형광세포화학현시열휴극도치αB-정상체단백재세포장내발생분포변화,이HSC70종포장향포핵이위;③면역전경정량분석진일보증실αB-정상체단백재열휴극시주요이향Z선급핵막등세포골가부위;④Northern-blot현시열휴극유도심기세포αB-정상체단백mRNA적표체증가。열휴극처리사αB-정상체단백신속이위지세포골가,표명기재심기예괄응조기상중가능발휘보호작용。이αB-정상체단백적유도표체,가능시심기예괄응연지상보호적중요궤제지일。
The effects of heat shock pretreatment on translocation andexpression of αB-crystallin were evaluated in neonatal rat cardiomyocytes. Results: ① Western blot analysis demonstrated that cytosolic soluble αB-crystallin rapidly translocated to insoluble intracellular structure at 10 or 30 minutes and returned to cytosolic soluble compartment at about 1~2h after heat shock. ② Double staining of immunofluorescence showed that αB-crystallin redistributed in cytosol after heat shock, while HSC70 translocated from cytosol to nucleus. ③ Quantitative analysis with immunoelectronic microscopy demonstrated αB-crystallin translocated to cytoskeletal structure such as Z-line and nuclear envelope after heat shock. ④ Northern-blot showed that heat shock induced αB-crystallin mRNA expression. The rapid translocation and increasing expression of αB-crystallin after heat shock suggests that this small heat shock protein may play an important protective roles during the early and late phases of myocardial ischemic preconditioning.