中国中西医结合急救杂志
中國中西醫結閤急救雜誌
중국중서의결합급구잡지
INTEGRATED TRADITIONAL CHINESE AND WESTERN MEDICINE IN PRACTICE OF CRITICAL CARE MEDICINE
2014年
2期
95-98
,共4页
任妙丹%何爱文%陈寿权%李章平%乔江华%李东芳%李惠萍%黄唯佳%程俊彦
任妙丹%何愛文%陳壽權%李章平%喬江華%李東芳%李惠萍%黃唯佳%程俊彥
임묘단%하애문%진수권%리장평%교강화%리동방%리혜평%황유가%정준언
微小RNA%心肺复苏%脑损伤%细胞凋亡
微小RNA%心肺複囌%腦損傷%細胞凋亡
미소RNA%심폐복소%뇌손상%세포조망
MicroRNA%Cardiopulmonary resuscitation%Brain injury%Apoptosis
目的：观察心搏骤停-心肺复苏（CA-CPR）后大鼠大脑皮质区凋亡相关性微小RNA（miRNA）的表达变化，探讨影响CPR的机制。方法选择成年雄性SD大鼠24只，按随机数字表法分为正常对照组、假手术组和CA-CPR组，每组8只。采用窒息致CA后CPR法建立CA-CPR大鼠模型；正常对照组不进行任何操作；假手术组仅施行腹腔麻醉、气管切开、血管穿刺及心电监护等操作，不夹闭气管，不复苏。正常对照组于正常饲养时、假手术组于气管切开后24 h时、CA-CPR组于自主循环恢复（ROSC）后各取大脑皮质区提取总RNA，采用实时荧光定量逆转录-聚合酶链反应（RT-PCR）测定大脑皮质区凋亡相关性miRNA的表达；采用流式细胞仪检测神经细胞凋亡率。结果与正常对照组比较，假手术组大鼠miRNA表达及细胞凋亡率差异无统计学意义（均P＞0.05）。与假手术组比较，CA-CPR组大鼠大脑皮质凋亡相关性miRNA有16个上调（分别为Let-7c，miR-15a，miR-21，miR-24，miR-29，miR-29b，miR-34a，miR-103，miR-200a，miR-200b， miR-200c，miR-210，miR-326，miR-338-3p，miR-494，miR-497），22个下调（分别为Let-7a，Let-7b，Let-7d， Let-7e，miR-19a，miR-19b-1，miR-20a，miR-20b，miR-23a，miR-23b，miR-25，miR-98，miR-107， miR-122a，miR-125a，miR-125b，miR-145，miR-181a，miR-181c，miR-335，miR-384-5p，miR-422a），其中有8个miRNA出现变化明显，miR-15a、miR-21、miR-34a、miR-497分别上调了（6.831±2.625）、（8.122±3.442）、（5.349±2.010）、（6.590±3.689）倍；miR-125b、miR-145、Let-7a、Let-7e分别下调了（0.122±0.039）、（0.199±0.096）、（0.191±0.069）、（0.160±0.082）倍。CA-CPR组皮质区细胞凋亡率〔（32.23±5.31）%〕较正常对照组〔（3.66±1.34）%〕和假手术组〔（4.98±1.84）%〕明显升高（均P＜0.01）。结论大鼠CA-CPR后早期脑组织存在明显的细胞凋亡，同期大脑皮质区凋亡相关性miRNA的表达也发生变化，其中发生明显变化的miRNA可能是CA-CPR后脑保护的新靶标。
目的：觀察心搏驟停-心肺複囌（CA-CPR）後大鼠大腦皮質區凋亡相關性微小RNA（miRNA）的錶達變化，探討影響CPR的機製。方法選擇成年雄性SD大鼠24隻，按隨機數字錶法分為正常對照組、假手術組和CA-CPR組，每組8隻。採用窒息緻CA後CPR法建立CA-CPR大鼠模型；正常對照組不進行任何操作；假手術組僅施行腹腔痳醉、氣管切開、血管穿刺及心電鑑護等操作，不夾閉氣管，不複囌。正常對照組于正常飼養時、假手術組于氣管切開後24 h時、CA-CPR組于自主循環恢複（ROSC）後各取大腦皮質區提取總RNA，採用實時熒光定量逆轉錄-聚閤酶鏈反應（RT-PCR）測定大腦皮質區凋亡相關性miRNA的錶達；採用流式細胞儀檢測神經細胞凋亡率。結果與正常對照組比較，假手術組大鼠miRNA錶達及細胞凋亡率差異無統計學意義（均P＞0.05）。與假手術組比較，CA-CPR組大鼠大腦皮質凋亡相關性miRNA有16箇上調（分彆為Let-7c，miR-15a，miR-21，miR-24，miR-29，miR-29b，miR-34a，miR-103，miR-200a，miR-200b， miR-200c，miR-210，miR-326，miR-338-3p，miR-494，miR-497），22箇下調（分彆為Let-7a，Let-7b，Let-7d， Let-7e，miR-19a，miR-19b-1，miR-20a，miR-20b，miR-23a，miR-23b，miR-25，miR-98，miR-107， miR-122a，miR-125a，miR-125b，miR-145，miR-181a，miR-181c，miR-335，miR-384-5p，miR-422a），其中有8箇miRNA齣現變化明顯，miR-15a、miR-21、miR-34a、miR-497分彆上調瞭（6.831±2.625）、（8.122±3.442）、（5.349±2.010）、（6.590±3.689）倍；miR-125b、miR-145、Let-7a、Let-7e分彆下調瞭（0.122±0.039）、（0.199±0.096）、（0.191±0.069）、（0.160±0.082）倍。CA-CPR組皮質區細胞凋亡率〔（32.23±5.31）%〕較正常對照組〔（3.66±1.34）%〕和假手術組〔（4.98±1.84）%〕明顯升高（均P＜0.01）。結論大鼠CA-CPR後早期腦組織存在明顯的細胞凋亡，同期大腦皮質區凋亡相關性miRNA的錶達也髮生變化，其中髮生明顯變化的miRNA可能是CA-CPR後腦保護的新靶標。
목적：관찰심박취정-심폐복소（CA-CPR）후대서대뇌피질구조망상관성미소RNA（miRNA）적표체변화，탐토영향CPR적궤제。방법선택성년웅성SD대서24지，안수궤수자표법분위정상대조조、가수술조화CA-CPR조，매조8지。채용질식치CA후CPR법건립CA-CPR대서모형；정상대조조불진행임하조작；가수술조부시행복강마취、기관절개、혈관천자급심전감호등조작，불협폐기관，불복소。정상대조조우정상사양시、가수술조우기관절개후24 h시、CA-CPR조우자주순배회복（ROSC）후각취대뇌피질구제취총RNA，채용실시형광정량역전록-취합매련반응（RT-PCR）측정대뇌피질구조망상관성miRNA적표체；채용류식세포의검측신경세포조망솔。결과여정상대조조비교，가수술조대서miRNA표체급세포조망솔차이무통계학의의（균P＞0.05）。여가수술조비교，CA-CPR조대서대뇌피질조망상관성miRNA유16개상조（분별위Let-7c，miR-15a，miR-21，miR-24，miR-29，miR-29b，miR-34a，miR-103，miR-200a，miR-200b， miR-200c，miR-210，miR-326，miR-338-3p，miR-494，miR-497），22개하조（분별위Let-7a，Let-7b，Let-7d， Let-7e，miR-19a，miR-19b-1，miR-20a，miR-20b，miR-23a，miR-23b，miR-25，miR-98，miR-107， miR-122a，miR-125a，miR-125b，miR-145，miR-181a，miR-181c，miR-335，miR-384-5p，miR-422a），기중유8개miRNA출현변화명현，miR-15a、miR-21、miR-34a、miR-497분별상조료（6.831±2.625）、（8.122±3.442）、（5.349±2.010）、（6.590±3.689）배；miR-125b、miR-145、Let-7a、Let-7e분별하조료（0.122±0.039）、（0.199±0.096）、（0.191±0.069）、（0.160±0.082）배。CA-CPR조피질구세포조망솔〔（32.23±5.31）%〕교정상대조조〔（3.66±1.34）%〕화가수술조〔（4.98±1.84）%〕명현승고（균P＜0.01）。결론대서CA-CPR후조기뇌조직존재명현적세포조망，동기대뇌피질구조망상관성miRNA적표체야발생변화，기중발생명현변화적miRNA가능시CA-CPR후뇌보호적신파표。
Objective To observe the expression changes in apoptosis-related microRNA(miRNA) in cerebral cortex after cardiac arrest-cardiopulmonary resuscitation(CA-CPR)in rats and explore the factors that may affect the mechanism of CPR. Methods 24 clean male Sprague-Dawley(SD)rats were randomly divided into three groups,the normal control group,sham operation group and CA-CPR group(each n=8). The animal model of CA induced by asphyxia was established and CPR was performed. In the normal control group,no special management was performed. In the sham operation group,only abdominal cavity anesthesia,tracheotomy,vascular puncture and electrocardiogram(ECG)were performed without clamping the trachea and resuscitating. Normal feeding in normal control group and 24 hours after tracheotomy in sham operation group,at 24 hours after recovery of spontaneous circulation(ROSC)in CA-CPR group,cerebral cortex specimens were obtained for detection of the expression of miRNA by using real time fluorescence quantitative reverse transcription - polymerase chain reaction(RT-PCR). Flow cytometry(FCM)was used to detect the neurocyte apoptotic rate. Results Compared between normal control and sham operation groups,there were no significant differences in the expression of apoptosis-related miRNA and neurocyte apoptosis rate of cerebral cortex(both P>0.05). Compared with sham operation group,in CA-CPR group, 16 miRNA expressions were up-regulated,including Let-7c,miR-15a,miR-21,miR-24,miR-29,miR-29b, miR-34a, miR-103, miR-200a, miR-200b, miR-200c, miR-210, miR-326, miR-338-3p, miR-494 and miR-497,and there were 22 down-regulated,being Let-7a,Let-7b,Let-7d,Let-7e,miR-19a,miR-19b-1, miR-20a,miR-20b,miR-23a,miR-23b,miR-25,miR-98,miR-107,miR-122a,miR-125a,miR-125b, miR-145,miR-181a,miR-181c,miR-335,miR-384-5p and miR-422a. Eight miRNA had significant changes at 24 hours after ROSC,in which miR-15a,miR-21,miR-34a,miR-497 were up-regulated respectively for 6.831±2.625,8.122±3.442,5.349±2.010,6.590±3.689 times,and miR-125b,miR-145,Let-7a,Let-7e were down-regulated respectively for 0.122±0.039,0.199±0.096,0.191±0.069,0.160±0.082 times. The apoptosis rate of cerebral cortex was increased significantly in CA-CPR group〔(32.23±5.31)%〕compared with that in normal control group〔(3.66±1.34)%〕and sham operation group〔(4.98±1.84)%,both P<0.01〕. Conclusions In early period after CA-CPR,obvious neurocyte apoptosis may be found in brain tissue of rats,and in the mean time, changes in apoptosis-related miRNA expression in cerebral cortex occur. The various types of miRNA with significant changes possibly play important roles in cerebral protection after CA-CPR in rats. <br> down-regulated respectively for 0.122±0.039,0.199±0.096,0.191±0.069,0.160±0.082 times. The apoptosis rate of cerebral cortex was increased significantly in CA-CPR group〔(32.23±5.31)%〕compared with that in normal control group〔(3.66±1.34)%〕and sham operation group〔(4.98±1.84)%,both P<0.01〕. Conclusions In early period after CA-CPR,obvious neurocyte apoptosis may be found in brain tissue of rats,and in the mean time, changes in apoptosis-related miRNA expression in cerebral cortex occur. The various types of miRNA with significant changes possibly play important roles in cerebral protection after CA-CPR in rats.
