中华显微外科杂志
中華顯微外科雜誌
중화현미외과잡지
Chinese Journal of Microsurgery
2014年
4期
368-372
,共5页
周明武%李琛琪%杨瑞甫%王广兰%罗彦平%王义生
週明武%李琛琪%楊瑞甫%王廣蘭%囉彥平%王義生
주명무%리침기%양서보%왕엄란%라언평%왕의생
缺血再灌注损伤%骨骼肌%Caveolin-3%小窝蛋白
缺血再灌註損傷%骨骼肌%Caveolin-3%小窩蛋白
결혈재관주손상%골격기%Caveolin-3%소와단백
Ischemia-reperfusion injury%Skeletal muscle%Caveolin-3%Caveolae
目的 观察兔肢体缺血再灌注损伤不同时相骨骼肌细胞损伤及Caveolin-3 mRNA的表达规律.方法 2013年4月至2013年12月,健康中国白兔15只(30只下肢),左下肢采用无创血管夹夹闭左侧髂总动脉,制成缺血再灌注模型作为实验组;右下肢不经夹闭血管处理作为对照组.分别于缺血5h、再灌注4h及8h取腓肠肌样本,采用HE染色法对骨骼肌组织进行处理,并通过光学显微镜观察;实时荧光定量PCR仪对Caveolin-3及GAPDH进行检测并采用-△△CT法计算Caveolin-3/GAPDH mRNA相对表达CT值. 结果 HE染色观察:对照组无水肿、变性及炎细胞浸润.实验组缺血5h肌细胞即发生变性、水肿,再灌注4h水肿加重、大量空泡形成及炎细胞浸润,再灌注8h较4h明显减轻.使用SPSS 19.0对Caveolin-3/GAPDH mRNA相对表达量统计分析:缺血5h、再灌注4h、再灌注8 h Caveolin-3/GAPDH mRNA相对表达量对照组分别为:1.026±0.065,1.004±0.037、1.022 ±0.051;实验组分别为:1.159±0.073、1.445±0.053和1.208±0.058.方差分析得出相应时间点实验组较对照组均增加,差异有统计学意义(P＜0.05).实验组缺血5h与再灌注8h比较差异无统计学意义(P＞0.05),缺血5h与再灌注4h、再灌注4h与再灌注8h比较差异有统计学意义(P＜0.05). 结论 骨骼肌细胞内Caveolin-3 mRNA表达量在肢体缺血再灌注损伤过程中较无损伤时增加,且缺血5h、再灌注4h、再灌注8h过程中Caveolin-3 mRNA表达量呈先增后减趋势.该现象同肌细胞损伤的发生发展过程相一致,提示Caveolin-3可能对骨骼肌细胞损伤及恢复具有某种调控作用.
目的 觀察兔肢體缺血再灌註損傷不同時相骨骼肌細胞損傷及Caveolin-3 mRNA的錶達規律.方法 2013年4月至2013年12月,健康中國白兔15隻(30隻下肢),左下肢採用無創血管夾夾閉左側髂總動脈,製成缺血再灌註模型作為實驗組;右下肢不經夾閉血管處理作為對照組.分彆于缺血5h、再灌註4h及8h取腓腸肌樣本,採用HE染色法對骨骼肌組織進行處理,併通過光學顯微鏡觀察;實時熒光定量PCR儀對Caveolin-3及GAPDH進行檢測併採用-△△CT法計算Caveolin-3/GAPDH mRNA相對錶達CT值. 結果 HE染色觀察:對照組無水腫、變性及炎細胞浸潤.實驗組缺血5h肌細胞即髮生變性、水腫,再灌註4h水腫加重、大量空泡形成及炎細胞浸潤,再灌註8h較4h明顯減輕.使用SPSS 19.0對Caveolin-3/GAPDH mRNA相對錶達量統計分析:缺血5h、再灌註4h、再灌註8 h Caveolin-3/GAPDH mRNA相對錶達量對照組分彆為:1.026±0.065,1.004±0.037、1.022 ±0.051;實驗組分彆為:1.159±0.073、1.445±0.053和1.208±0.058.方差分析得齣相應時間點實驗組較對照組均增加,差異有統計學意義(P＜0.05).實驗組缺血5h與再灌註8h比較差異無統計學意義(P＞0.05),缺血5h與再灌註4h、再灌註4h與再灌註8h比較差異有統計學意義(P＜0.05). 結論 骨骼肌細胞內Caveolin-3 mRNA錶達量在肢體缺血再灌註損傷過程中較無損傷時增加,且缺血5h、再灌註4h、再灌註8h過程中Caveolin-3 mRNA錶達量呈先增後減趨勢.該現象同肌細胞損傷的髮生髮展過程相一緻,提示Caveolin-3可能對骨骼肌細胞損傷及恢複具有某種調控作用.
목적 관찰토지체결혈재관주손상불동시상골격기세포손상급Caveolin-3 mRNA적표체규률.방법 2013년4월지2013년12월,건강중국백토15지(30지하지),좌하지채용무창혈관협협폐좌측가총동맥,제성결혈재관주모형작위실험조;우하지불경협폐혈관처리작위대조조.분별우결혈5h、재관주4h급8h취비장기양본,채용HE염색법대골격기조직진행처리,병통과광학현미경관찰;실시형광정량PCR의대Caveolin-3급GAPDH진행검측병채용-△△CT법계산Caveolin-3/GAPDH mRNA상대표체CT치. 결과 HE염색관찰:대조조무수종、변성급염세포침윤.실험조결혈5h기세포즉발생변성、수종,재관주4h수종가중、대량공포형성급염세포침윤,재관주8h교4h명현감경.사용SPSS 19.0대Caveolin-3/GAPDH mRNA상대표체량통계분석:결혈5h、재관주4h、재관주8 h Caveolin-3/GAPDH mRNA상대표체량대조조분별위:1.026±0.065,1.004±0.037、1.022 ±0.051;실험조분별위:1.159±0.073、1.445±0.053화1.208±0.058.방차분석득출상응시간점실험조교대조조균증가,차이유통계학의의(P＜0.05).실험조결혈5h여재관주8h비교차이무통계학의의(P＞0.05),결혈5h여재관주4h、재관주4h여재관주8h비교차이유통계학의의(P＜0.05). 결론 골격기세포내Caveolin-3 mRNA표체량재지체결혈재관주손상과정중교무손상시증가,차결혈5h、재관주4h、재관주8h과정중Caveolin-3 mRNA표체량정선증후감추세.해현상동기세포손상적발생발전과정상일치,제시Caveolin-3가능대골격기세포손상급회복구유모충조공작용.
Objective To observe the damage degree and expression pattern of Caveolin-3 mRNA by ischemia-reperfusion injury in rabbits of skeletal muscle cell at different phases.Methods In this study,from April 2013 to December 2013,30 lower limbs of 15 Chinese White Rabbits were used and divided into two groups:all the left lower limbs were experimental group,which were made as an experimental model of ischemia-reperfusion injury by occluding left common iliac artery using noninvasive vascular.All the right lower limbs without surgical treatment were the control group.Gastrocnemius samples were obtained at 4h and 8h after reperfusion and handled by HE staining and observed by optical microscopy.By Real-time PCR,Caveolin-3/GAPDH mRNA were detected.Results HE stain showed:in control group,there was no edema,degeneration and inflammatory cell infiltration; in experi-meatal group,muscle cell degeneration had occured at ischemic 5 h.The edema was aggravated,a large number vacuole were formed and inflammatory cell were infiltrated at 4 h reperfusion.Reperfusion injury at 8h significantly reduced compared to 4 h.The Caveolin-3/GAPDH mRNA expression levels by SPSS 19.0 showed:Control group:1.026 ± 0.065,1.004 ±0.037,1.022 ±0.051,experimental group:1.159 ±0.073,1.445 ±0.053,1.208 ±0.058 at ischemic 5 h,4 h and 8 h reperfusion,respectively.On-line analysis of variance cases of ischemic 5 h and 4 h reperfusion and 8 h reperfusion,the experimental group than the control group were increased,with statistical significance (P ＜ 0.05).The experimental group of ischemic 5 h and 8 h reperfusion was no significant difference (P ＞ 0.05).It showed Caveolin-3 mRNA expression levels in ischemia-reperfusion 8 h group returned to normal.There was significant statistical difference between the ischemic 5 h and 4 h reperfusion (P ＜ 0.05).There was significant statistical difference between the 4 h reperfusion and 8 h reperfusion (P ＜ 0.05).Conclusion The expression of Caveolin-3 in experimental group showed a trend of first increased and then decreased.The expression levels of Caveolin-3 mRNA in skeletal muscle cells after ischemia-reperfusion injury is consistent with the development and progression of muscle cell damage.The results indicate that Caveolin-3 may play a control role in the injury and recovery of skeletal muscle cell.
