ISSN 0371-0874, CN 31-1352/Q



潘燕霞, 林丽, 袁文俊, 吴国宏, 唐朝枢



实验观察了 01一1 nmol/ L内皮素一1 (ET-1)预处理对低氧孵育((3 % 02 -5 % C02, 12 h)的培养乳鼠心肌细胞乳酸脱氢酶(LDH)释放量、培养液上清超氧化物歧化酶(SOD)活性以及丙二醛( MDA)含量的影响。用Fluo3/ AM负载培养的心肌细胞,在激光扫描共聚焦显微镜下监测急性低氧的心肌细胞「Ca~(2+)];的变化和ET1预处理对低氧所致「Ca~(2+)];变化的影响。结果如下:(1)心肌细胞低氧孵育12 h后,培养液上清LDH活力和MDA含量较常氧对照组明显升高,分别为43. 33士1.21 U/ L二19.33士1. 03 U/ L和1.71士0. 02 nmol/ L二0.91士0. 03 nmol/ L ( PX0.01),SOD活性为16. 93士1.11 U/ ml明显低于常氧对照组的33. 48士1.15 U/ ml (P<0.01);0.01一1 nmol/ LET-1预处理呈浓度依赖性抑制低氧培养心肌细胞LDH释放,减少培养液上清MDA含量、提高SOD活性(P<0.01) o (2)低氧灌流后29士1. 5 s ( n = 23)心肌细胞自发性钙瞬变完全终止,[ Ca~(2+) ];升高了107士3.2% ( P<0. 001) ; 0. 01一1 nmol/ L ET-1能明显加快心肌细胞钙瞬变的频率(P<0.01);ET-1预处理后低氧所致钙瞬变终止的时间较单纯低氧组明显推迟,[ Ca~(2+)+];过度升高被明显减轻(P<0.01)。上述结果表明,0.01一1 nmol/ L ET-1预处理可减轻培养乳鼠心肌细胞的低氧损伤和抑制低氧所致「Ca~(2+)];的变化,具有一定的细胞保护作用。

关键词: 内皮素; 低氧; 心肌细胞; 细胞内钙

Preventive effect of endothelia 1 pretreatment on hypoxix induced injury in cultured neonatal rat cardiomyoc尹e

PAN Yan Xia, Lin Li, YUAN Weir Jun, WU Guo-Hong, TANG CHAO-SHU

'Department of Physiology, Second Military Medical University, Shanghai 200433;Department of Physiology, Fujian Medical University, Fuzhou 350004;Department of Physiology and Pathophysiology,Health Sciences Center,Peking University,Beijing 100083


This study was designed to observe the effects of endothelixrl(ET-1)pretreatment on hypoxia7induced injury and changes in[Ca~(2+)],in cultured neonatal rat cardiomyocytes .The activity of lactate dehydrogenase(LDH) and superoxide dis mutase(S OD),and the content of malondialdehyde
(MDA) in the supernatant were determined in the cultured cardiomyocytes su匀ected to a 12-h hypoxia induced by a 3q一5 % Cq atmosphere at 37℃with or without ET-1 pretreatment.[Ca~(2+)],was measured with Ca~(2+)sensitive fluorescent probe fluo-3/ AM under a laser scanning confocal microscope .Fluo-
rescence intensity emitted from fluo-3/ AM-loaded cells reflected the concentration of[Ca~(2+)],.The hypoxia model used in[Ca~(2+)],measurement was established by continously perfusing cardiomyocytes for 30 min with 95%NZ 5%CO2 saturated D ME M solution containing 1 m mol/ L Na2S2 04.Pretreatment with ET-1 consisted of three cycles of ET-1 perfusion(5 min for each) followed by E T-1一free DME M solution (1 0 min for each) prior to hypoxia .The results showed that(1)after incubation in a 3%02 -5%C02 hypoxic atmosphere for 1 2 h,the activity of LDH and the content of MDA in the supernatant significant-
1y increase dfrom 19.33士1.03 U/ L to 43. 33士1.21 U/ L and from 0. 91士0. 03 nmol/ L to 1.71士0. 02nmol/ L,respectively,whereas the activity of SOD decreased from 33.48士1.15 U/ ml to 16. 93士1.11U/ ml(P<0. 01)·In hypoxic cardiomyocytes pretreated with 0. 01一1 nmol/ L ET-1,LDH release and
supernatant MDA content were decreased,while SOD activity was enhanced dose-dependently(P<0. 01).(2) The spontaneous calcium transient in cultured cardiomyocytes terminated at 29士1.5 s and [Cat十],increased by 107士13.2 % during perfusion of hypoxic solution(P<0. 001)at the end of 30 min .ET-1(0. 01一1 nmol/ L) increased the frequency of[Ca~(2+)],transient in cultured cardiomyocytes in a dose-dependent manner(P<0.01).The termination of[Ca~(2+)],transient and the elevation of[Cat十]1caused by hypoxia were postponed by pretreatment with 0. 01一1 nmol/ L ET -I(P<0. 01).These results show that pretreatment with 0. 01一1 nmol/ L ET-1 attenuates hypoxia7induced injury and[Cat十]1changes in cultured neonatal cardiomyocytes,indicating a cytoprotective role of ET-1 pretreatment.

Key words: hypoxia;cardiomyocytes;intracellular calcium ;

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潘燕霞, 林丽, 袁文俊, 吴国宏, 唐朝枢. 内皮-1预处理对培养乳鼠心肌细胞低氧损伤的保护作用[J]. 生理学报 2003; 55 (2): .

PAN Yan Xia, Lin Li, YUAN Weir Jun, WU Guo-Hong, TANG CHAO-SHU. Preventive effect of endothelia 1 pretreatment on hypoxix induced injury in cultured neonatal rat cardiomyoc尹e. Acta Physiol Sin 2003; 55 (2): (in Chinese with English abstract).