中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2011年
4期
295-298
,共4页
黄邵洪%荣健%刘子由%刘海%吴钟凯
黃邵洪%榮健%劉子由%劉海%吳鐘凱
황소홍%영건%류자유%류해%오종개
再灌注损伤%钙离子转运通道蛋白%心肌%细胞膜%RNA,信使
再灌註損傷%鈣離子轉運通道蛋白%心肌%細胞膜%RNA,信使
재관주손상%개리자전운통도단백%심기%세포막%RNA,신사
Reperfusion injury%Calcium transporting channel proteins%Myocardium%Cell membrane%RNA,messenger
目的 探讨大鼠心肌在体缺血再灌注(IR)损伤后细胞膜钙转运通道蛋白的mRNA变化对钙超载的作用。方法 12只SD大鼠按随机数字法分为IR组和对照组。IR组通过结扎（缺血20 min）后松解(再灌注60 min)前降支造成心肌IR,对照组则免除结扎松解前降支。应用生理记录仪连续监测两组大鼠缺血开始前及再灌注60 min后心率、平均动脉压等血流动力学指标。全自动生化仪检测缺血前及再灌注60 min后两组大鼠血钙及肌钙蛋白T(cTnT)的水平。荧光定量PCR检测再灌注60 min后两组大鼠左心室缺血区和右心室心肌细胞膜钙转运通道蛋白即心肌细胞膜钠钙交换器1(NCX1)，L型钙通道(LVDCC)α-1C和胞膜钙转运ATP酶1(PMCA1 )mRNA的表达。结果两组大鼠缺血前的心率、平均动脉压均高于再灌注60 min后，而两组间缺血前和再灌注60 min后的心率、平均动脉压差异则无统计学意义。两组血浆Ca2+浓度在缺血前与再灌注60 min后差异无统计学意义，同时间点两组之间的差异也没有统计学意义。缺血前IR组与对照组血浆cTnT浓度水平相近，缺血60 min后IR组血浆cTnT浓度较对照组升高[(4.29±2.22) μg/L比(1.62±0.60)μg/L，P=0.031]；两组血浆cTnT浓度在缺血前与再灌注60 min后差异也有统计学意义（均P＜0.05）。NCX1，LVDCCα-1C和PMCA1的mRNA表达在再灌注60 min后同心室两组间和同组内左右心室之间的差异均无统计学意义（NCX1：对照组左心室为50±4，右心室为47±9；IR组左心室为55±6,右心室为53±11；LVDCCα-1C:对照组左心室为33±7，右心室为30±7；IR组左心室为28±3，右心室为37±5；PMCA1，对照组左心室为70±10,右心室为53±11；IR组左心室为66±12，右心室为78±8;均P＞0.05）。结论 大鼠在体心肌缺血20 min再灌注60 min后,NCX1、LVDCCα-1C和PMCA1的mRNA表达水平均无显著改变，提示钙超载并非由细胞膜钙转运通道蛋白数量改变引起。
目的 探討大鼠心肌在體缺血再灌註(IR)損傷後細胞膜鈣轉運通道蛋白的mRNA變化對鈣超載的作用。方法 12隻SD大鼠按隨機數字法分為IR組和對照組。IR組通過結扎（缺血20 min）後鬆解(再灌註60 min)前降支造成心肌IR,對照組則免除結扎鬆解前降支。應用生理記錄儀連續鑑測兩組大鼠缺血開始前及再灌註60 min後心率、平均動脈壓等血流動力學指標。全自動生化儀檢測缺血前及再灌註60 min後兩組大鼠血鈣及肌鈣蛋白T(cTnT)的水平。熒光定量PCR檢測再灌註60 min後兩組大鼠左心室缺血區和右心室心肌細胞膜鈣轉運通道蛋白即心肌細胞膜鈉鈣交換器1(NCX1)，L型鈣通道(LVDCC)α-1C和胞膜鈣轉運ATP酶1(PMCA1 )mRNA的錶達。結果兩組大鼠缺血前的心率、平均動脈壓均高于再灌註60 min後，而兩組間缺血前和再灌註60 min後的心率、平均動脈壓差異則無統計學意義。兩組血漿Ca2+濃度在缺血前與再灌註60 min後差異無統計學意義，同時間點兩組之間的差異也沒有統計學意義。缺血前IR組與對照組血漿cTnT濃度水平相近，缺血60 min後IR組血漿cTnT濃度較對照組升高[(4.29±2.22) μg/L比(1.62±0.60)μg/L，P=0.031]；兩組血漿cTnT濃度在缺血前與再灌註60 min後差異也有統計學意義（均P＜0.05）。NCX1，LVDCCα-1C和PMCA1的mRNA錶達在再灌註60 min後同心室兩組間和同組內左右心室之間的差異均無統計學意義（NCX1：對照組左心室為50±4，右心室為47±9；IR組左心室為55±6,右心室為53±11；LVDCCα-1C:對照組左心室為33±7，右心室為30±7；IR組左心室為28±3，右心室為37±5；PMCA1，對照組左心室為70±10,右心室為53±11；IR組左心室為66±12，右心室為78±8;均P＞0.05）。結論 大鼠在體心肌缺血20 min再灌註60 min後,NCX1、LVDCCα-1C和PMCA1的mRNA錶達水平均無顯著改變，提示鈣超載併非由細胞膜鈣轉運通道蛋白數量改變引起。
목적 탐토대서심기재체결혈재관주(IR)손상후세포막개전운통도단백적mRNA변화대개초재적작용。방법 12지SD대서안수궤수자법분위IR조화대조조。IR조통과결찰（결혈20 min）후송해(재관주60 min)전강지조성심기IR,대조조칙면제결찰송해전강지。응용생리기록의련속감측량조대서결혈개시전급재관주60 min후심솔、평균동맥압등혈류동역학지표。전자동생화의검측결혈전급재관주60 min후량조대서혈개급기개단백T(cTnT)적수평。형광정량PCR검측재관주60 min후량조대서좌심실결혈구화우심실심기세포막개전운통도단백즉심기세포막납개교환기1(NCX1)，L형개통도(LVDCC)α-1C화포막개전운ATP매1(PMCA1 )mRNA적표체。결과량조대서결혈전적심솔、평균동맥압균고우재관주60 min후，이량조간결혈전화재관주60 min후적심솔、평균동맥압차이칙무통계학의의。량조혈장Ca2+농도재결혈전여재관주60 min후차이무통계학의의，동시간점량조지간적차이야몰유통계학의의。결혈전IR조여대조조혈장cTnT농도수평상근，결혈60 min후IR조혈장cTnT농도교대조조승고[(4.29±2.22) μg/L비(1.62±0.60)μg/L，P=0.031]；량조혈장cTnT농도재결혈전여재관주60 min후차이야유통계학의의（균P＜0.05）。NCX1，LVDCCα-1C화PMCA1적mRNA표체재재관주60 min후동심실량조간화동조내좌우심실지간적차이균무통계학의의（NCX1：대조조좌심실위50±4，우심실위47±9；IR조좌심실위55±6,우심실위53±11；LVDCCα-1C:대조조좌심실위33±7，우심실위30±7；IR조좌심실위28±3，우심실위37±5；PMCA1，대조조좌심실위70±10,우심실위53±11；IR조좌심실위66±12，우심실위78±8;균P＞0.05）。결론 대서재체심기결혈20 min재관주60 min후,NCX1、LVDCCα-1C화PMCA1적mRNA표체수평균무현저개변，제시개초재병비유세포막개전운통도단백수량개변인기。
Objective To investigate the effect of altered mRNA expression of sarcolemmal calcium transporting channel proteins in rat myocardial ischemia reperfusion injury (IR)in vivo on calcium overload.Methods Twelve SD rats were randomized into IR and control groups. LAD coronary artery was ligated for 20 min (ischemia) then released for 60 min (repeffusion) in IR group but not in the control group. In both groups,physiologic recorder was used for continuous measurement of bemodynamic parameters including heart rote and mean arterial pressure; full-automated biochemical analyzer was used to determine the serum calcium and cTnT before ischemia and at 60 min after repedusion; quantitative PCR was used to detect the mRNA expression of sarcolemmal calcium transporting channel proteins (NCX 1, LVDCCα-1C and PMCAI ) in cardiomyocytes of the left ventricle (LV, isehemic area) and right ventricle (RV) at 60 min after reperfusion. Results Two groups did not differ in heart rate and mean arterial pressure either before ischemia or at 60 min after reperfusion, but both showed a reduction in these two parameters at 60 min after reperfusion compared with these before ischemia.There was no statistical difference of two groups in serum calcium before ischemia or at 60 min after reperfusion,or at any given time points between two groups. While the plasma cTnT level was comparable before ischemia between two groups. IR group showed significantly elevated cTnT at 60 min after reperfusion compared with the control group[(4.29±2.22) μg/L vs (1.62±0.60) μg/L, P=0.031]. There was statistical difference of two groups in plasma cTnT level before ischemia compared with this at 60 min after reperfusion(all P＜0.05). At 60 min after reperfusion, there were no differences in mRNA expression of sarcolemmal NCXI, LVDCCα-1C and PMCA1 of same ventricle between two groups or between two ventricles of same group (NCX1 : control group LV 50±4,control group RV 47±9; IR group LV 55±6, IR group RV 53±11 ; LVDCCα-1C: control group LV 33±7, control group RV 30±7; IR group LV 28±3, IR group RV 37±5; PMCA1 : control group LV 70±10, control group RV 53± 11 ; IR group LV 66±12, IR group RV 78±8; all P＞0.05). Conclusion The mRNA expression of NCX,LVDCCα-1C and PMCA1 does not change obviously at 20 min after ischemia and at 60 min after reperfusion in rat heart in vivo, suggesting that the quantity of sarcolemmal calcium transporting channel proteins may not be responsible for calcium overload.
