摘要
目的 探讨细胞内游离钙 [Ca2 + ]i在柯萨奇病毒 B3(CVB3)诱导培养心肌细胞凋亡中的作用。方法 DNA裂点检测法 (3′-末端标记 )及透射电镜检测细胞凋亡。 Fluo3- AM负载心肌细胞 ,共聚焦显微镜观察[Ca2 + ]i荧光强度变化。结果 感染 2 4h心肌细胞内 CVB3滴度达峰值。感染 10 h未见凋亡的心肌细胞 ,17、2 4和 36 h凋亡细胞分别为 5 %、6 0 %和 90 % ,感染 17h心肌 [Ca2 + ]i浓度达峰值。电镜结果提示 CVB3感染组存在典型的凋亡心肌细胞。结论 CVB3可诱导培养心肌细胞的凋亡过程。细胞内 Ca2 + 参与凋亡细胞的信息传导过程 。
Objective To explore the role of cytosolic free calcium ([Ca 2+ ]i) in apoptosis induced by coxsackievirus B_3 (CVB_3) in cultured cardiomyocytes of rats.Methods Primary cultured cardiomyocyte was prepared from Wistar rats ages 2~3 d.The apoptosis in cardiomyocyte was determined by terminated deoxynucleotide transferase directed d-UTP nick and end labeling (TUNEL) method,and the apoptosis was observed under a transmission electron microscope.[Ca 2+ ]i in single cardiomyocyte loaded with Fluo3-AM was measured by confocal microscope.Results The concentration of CVB_3 in the medium reached the peak at 24 h after CVB_3 infection.The apoptotic cells were not found in CVB_3 -infected cardiomyocyte in the first 10 h,but amounted to 5% at 17 h,60% at 24 h,and 90% at 36 h.The peak value of [Ca 2+ ]i elevation was reached at 17 h after CVB_3 infection (P<0.01) .The characteristics of apoptosis were also seen by transmission electron microscope.Conclusions CVB_3 induced the apoptosis in cultured cardiomyocyte,and [Ca 2+ ]i mobilization was involved in the signal transduction process in apoptosis cells,and played an important role especially in the early stage of apoptosis indued by CVB_3
出处
《中国地方病学杂志》
CAS
CSCD
2000年第4期248-250,共3页
Chinese Jouranl of Endemiology
基金
黑龙江省自然科学基金!( D9718)
关键词
柯萨奇病毒B3
细胞凋亡
钙
病毒性心肌炎
大鼠
Coxsackieviruses B_3
Apoptosis
Calcium
Confocal microscopy
Myocaridum cells
