中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2014年
8期
862-865
,共4页
潘志国%邵玉%万军%耿焱%陈镜合%苏磊
潘誌國%邵玉%萬軍%耿焱%陳鏡閤%囌磊
반지국%소옥%만군%경염%진경합%소뢰
热打击%骨骼肌细胞%通透性%钙超载%细胞骨架%细胞周期%考马斯亮蓝染色%流式检测
熱打擊%骨骼肌細胞%通透性%鈣超載%細胞骨架%細胞週期%攷馬斯亮藍染色%流式檢測
열타격%골격기세포%통투성%개초재%세포골가%세포주기%고마사량람염색%류식검측
Heat stress%Skeletal muscle cells%Permeability%Calcium overload%Cytoskeleton%Cell cycles%Coomassie brilliant blue%Coomassie brilliant blue%Flow cytometry
目的 研究热打击对培养人骨骼肌细胞(HSKMC)通透性、细胞骨架及细胞周期影响.方法 流式细胞仪钙离子内流检测热打击对HSKMC细胞膜通透性的影响,考马斯亮蓝R-250染色法检测热打击对HSKMC细胞骨架影响,流式细胞仪检测热打击对HSKMC细胞周期改变.结果 给予培养人HSKMC细胞不同温度梯度热打击1h后,43℃热打击组钙离子流式检测的中位数为91.63,37℃对照组中位数为22.98.同对照组相比,随着热打击程度加强,显微镜高倍镜下可见骨骼肌细胞骨架逐渐出现变粗、变短、出现明显的应力纤维.骨骼肌细胞在热打击情况下,各组G0/G1期DNA含量在44.13～62.98之间,与正常对照组对比,当细胞离开热打击环境后培养18 h前G0/G1期DNA含量明显高于对照组,培养第18小时恢复才同对照组基本相同.结论 热打击的情况下造成细胞钙离子内流效应,导致细胞内钙超载.可改变HSKMC骨架结构,使骨架失去正常的网状有序排列结构,间隙增大.细胞周期出现阻滞,细胞被阻滞在G0/G1期.
目的 研究熱打擊對培養人骨骼肌細胞(HSKMC)通透性、細胞骨架及細胞週期影響.方法 流式細胞儀鈣離子內流檢測熱打擊對HSKMC細胞膜通透性的影響,攷馬斯亮藍R-250染色法檢測熱打擊對HSKMC細胞骨架影響,流式細胞儀檢測熱打擊對HSKMC細胞週期改變.結果 給予培養人HSKMC細胞不同溫度梯度熱打擊1h後,43℃熱打擊組鈣離子流式檢測的中位數為91.63,37℃對照組中位數為22.98.同對照組相比,隨著熱打擊程度加彊,顯微鏡高倍鏡下可見骨骼肌細胞骨架逐漸齣現變粗、變短、齣現明顯的應力纖維.骨骼肌細胞在熱打擊情況下,各組G0/G1期DNA含量在44.13～62.98之間,與正常對照組對比,噹細胞離開熱打擊環境後培養18 h前G0/G1期DNA含量明顯高于對照組,培養第18小時恢複纔同對照組基本相同.結論 熱打擊的情況下造成細胞鈣離子內流效應,導緻細胞內鈣超載.可改變HSKMC骨架結構,使骨架失去正常的網狀有序排列結構,間隙增大.細胞週期齣現阻滯,細胞被阻滯在G0/G1期.
목적 연구열타격대배양인골격기세포(HSKMC)통투성、세포골가급세포주기영향.방법 류식세포의개리자내류검측열타격대HSKMC세포막통투성적영향,고마사량람R-250염색법검측열타격대HSKMC세포골가영향,류식세포의검측열타격대HSKMC세포주기개변.결과 급여배양인HSKMC세포불동온도제도열타격1h후,43℃열타격조개리자류식검측적중위수위91.63,37℃대조조중위수위22.98.동대조조상비,수착열타격정도가강,현미경고배경하가견골격기세포골가축점출현변조、변단、출현명현적응력섬유.골격기세포재열타격정황하,각조G0/G1기DNA함량재44.13～62.98지간,여정상대조조대비,당세포리개열타격배경후배양18 h전G0/G1기DNA함량명현고우대조조,배양제18소시회복재동대조조기본상동.결론 열타격적정황하조성세포개리자내류효응,도치세포내개초재.가개변HSKMC골가결구,사골가실거정상적망상유서배렬결구,간극증대.세포주기출현조체,세포피조체재G0/G1기.
Objective To study the effect of heat stress on the permeability,cytoskeleton and cell cycle of human skeletal muscle cell (HSKMC).Methods The HSKMC membrane permeability was detected by calcium ion inflow with flow cytometer,the cytoskeleton was stained by CBB 250,and the cell cycle was determined by flow cytometer.Results After 1 h of heat stress on human HSKMC cells under different temperature gradient,the median level of calcium ion was 91.63 in 43 ℃ heat stress group compared with 22.98 in 37 ℃ control group.As temperature increased,thicker and shorter cytoskeleton and stress fiber were shown under the high power lens of microscope.The DNA expression of skeleton cells at G0/G1 stage was 44.13-62.98 in groups under heat stress.Compared with normal control group,DNA expression was much higher in heat stress group,when HSKMC was cultured under 37 ℃ temperature for another 18 h,it kept decreasing DNA expression to a similar level as control group.Conclusions Heat stress can cause calcium iron inflow resulting in intracellular calcium overload,and affect the cytoskeleton leading to loss of normal web ordered arrangement and increased gap in HSKMC cells,which give rise to blocking cell cycle into G0/G1 stage.
