电子显微学报
電子顯微學報
전자현미학보
JOURNAL OF CHINESE ELECTRON MICROSCOPY SOCIETY
2015年
1期
55-61
,共7页
沈亚峰%袁青宁%汤莹%雷长海%范晓燕%林方兴%张玉霞%杨勇骥
瀋亞峰%袁青寧%湯瑩%雷長海%範曉燕%林方興%張玉霞%楊勇驥
침아봉%원청저%탕형%뢰장해%범효연%림방흥%장옥하%양용기
心力衰竭%钙瞬变%SERCA2a%PLN%PP1%激光扫描共聚焦显微技术
心力衰竭%鈣瞬變%SERCA2a%PLN%PP1%激光掃描共聚焦顯微技術
심력쇠갈%개순변%SERCA2a%PLN%PP1%격광소묘공취초현미기술
heart failure%calcium transients%SERCA2a%PLN%PP1%laser scanning confocal microscopy
目的:探讨心力衰竭时心肌细胞肌浆网钙回摄异常的机制。方法:结扎大鼠冠状动脉左前降支制备慢性心衰模型。4周后采用酶解法分离假手术组和心衰组大鼠心室肌细胞,利用激光扫描共聚焦显微镜检测心肌细胞肌浆网钙含量。采用western?blot技术检测心肌细胞肌浆网钙调蛋白SERCA2a、PLN、PP1的表达水平。结果:心衰组的大鼠心肌细胞荧光强度小于假手术组,并且心衰组大鼠心肌细胞咖啡因诱导的钙瞬变也低于假手术组;心衰组大鼠心SERCA2a表达水平降低,PP1的水平升高,PLN总体水平不变,但磷酸化的PLN水平降低。结论:钙泵表达量下降和钙泵活性降低是心力衰竭时心肌细胞肌浆网钙回摄减少的主要原因。
目的:探討心力衰竭時心肌細胞肌漿網鈣迴攝異常的機製。方法:結扎大鼠冠狀動脈左前降支製備慢性心衰模型。4週後採用酶解法分離假手術組和心衰組大鼠心室肌細胞,利用激光掃描共聚焦顯微鏡檢測心肌細胞肌漿網鈣含量。採用western?blot技術檢測心肌細胞肌漿網鈣調蛋白SERCA2a、PLN、PP1的錶達水平。結果:心衰組的大鼠心肌細胞熒光彊度小于假手術組,併且心衰組大鼠心肌細胞咖啡因誘導的鈣瞬變也低于假手術組;心衰組大鼠心SERCA2a錶達水平降低,PP1的水平升高,PLN總體水平不變,但燐痠化的PLN水平降低。結論:鈣泵錶達量下降和鈣泵活性降低是心力衰竭時心肌細胞肌漿網鈣迴攝減少的主要原因。
목적:탐토심력쇠갈시심기세포기장망개회섭이상적궤제。방법:결찰대서관상동맥좌전강지제비만성심쇠모형。4주후채용매해법분리가수술조화심쇠조대서심실기세포,이용격광소묘공취초현미경검측심기세포기장망개함량。채용western?blot기술검측심기세포기장망개조단백SERCA2a、PLN、PP1적표체수평。결과:심쇠조적대서심기세포형광강도소우가수술조,병차심쇠조대서심기세포가배인유도적개순변야저우가수술조;심쇠조대서심SERCA2a표체수평강저,PP1적수평승고,PLN총체수평불변,단린산화적PLN수평강저。결론:개빙표체량하강화개빙활성강저시심력쇠갈시심기세포기장망개회섭감소적주요원인。
To explore the mechanism of sarcoplasmic reticulum calcium uptake in heart failure myocytes. Methods: Adult rat ventricular myocytes were isolated by enzymatic digestion from the rat model of chronic heart failure, which was produced by ligation of the left anterior descending coronary artery. Sarcoplasmic reticulum calcium content was measured by laser scanning confocal microscopy;western blot method was used to analyze the differences in expression of calcium handling proteins in sham and heart failure. Results:The fluorescence intensity of ratsmyocytesin heart failure group was much lower than the sham group, and the caffeine induced calcium transients in heart failure group was lower than that of the sham group. The expression of PLN phosphorylation level decreased, but the level of overall PLN did not change. The expression of SERCA2a decreased and the level of PP1 increased. Conclusion:The main reasons for reducing sarcoplasmic reticulum calcium uptake were the reduction of SERCA2a expression and the decrease of SERCA2a activity.