内皮素-1预处理对培养乳鼠心肌细胞低氧损伤的保护作用  

Preventive effect of endothelin-1 pretreatment on hypoxia-induced injury in cultured neonatal rat cardiomyocytes

作　　者：潘燕霞[1] 林丽[1] 袁文俊[1] 吴国宏[1] 唐朝枢[2] 

机构地区：[1]第二军医大学生理学教研室 [2]北京大学医学部生理和病理生理学系,北京100083

出　　处：《生理学报》2003年第2期171-176,共6页Acta Physiologica Sinica

基　　金：ThisstudywassupportedbygrantsfromtheMajorStateBasicResearchDevelopmentProgramofthePeople'sRepublicofChina(No G2 0 0 0 0 5 690 5 )andtheNationalNaturalScienceFoundationofChina (No 30 0 70 30 6)

摘　　要：实验观察了 0 0 1- 1nmol/L内皮素 1(ET 1)预处理对低氧孵育 ( 3 %O2 5 %CO2 ,12h)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Fluo 3 /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 [Ca2 +]i 的变化和ET 1预处理对低氧所致 [Ca2 +]i 变化的影响。结果如下 :( 1)心肌细胞低氧孵育 12h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 3 3± 1 2 1U/Lvs 19 3 3± 1 0 3U/L和 1 71± 0 0 2nmol/Lvs 0 91± 0 0 3nmol/L (P<0 0 1) ,SOD活性为 16 93± 1 11U/ml明显低于常氧对照组的 3 3 48± 1 15U/ml (P <0 0 1) ;0 0 1- 1nmol/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 0 1)。 ( 2 )低氧灌流后 2 9± 1 5s (n =2 3 )心肌细胞自发性钙瞬变完全终止 ,[Ca2 +]i 升高了 10 7± 13 2 % (P <0 0 0 1) ;0 0 1- 1nmol/LET 1能明显加快心肌细胞钙瞬变的频率 (P <0 0 1) ;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 ,[Ca2 +]i过度升高被明显减轻 (P <0 0 1)。上述结果表明 ,0 0 1- 1nmol/LETThis study was designed to observe the effects of endothelin-1 (ET-1) pretreatment on hypoxia-induced injury and changes in i in cultured neonatal rat cardiomyocytes. The activity of lactate dehydrogenase (LDH) and superoxide dismutase (SOD), and the content of malondialdehyde (MDA) in the supernatant were determined in the cultured cardiomyocytes subjected to a 12-h hypoxia induced by a 3% O 2-5% CO 2 atmosphere at 37℃ with or without ET-1 pretreatment. i was measured with Ca 2+-sensitive fluorescent probe fluo-3/AM under a laser scanning confocal microscope. Fluorescence intensity emitted from fluo-3/AM-loaded cells reflected the concentration of i. The hypoxia model used in i measurement was established by continously perfusing cardiomyocytes for 30 min with 95% N 2-5% CO 2 saturated DMEM solution containing 1 mmol/L Na 2S 2O 4. Pretreatment with ET-1 consisted of three cycles of ET-1 perfusion (5 min for each) followed by ET-1-free DMEM solution (10 min for each) prior to hypoxia. The results showed that (1) after incubation in a 3% O 2-5% CO 2 hypoxic atmosphere for 12 h, the activity of LDH and the content of MDA in the supernatant significantly increased from 19 33±1 03 U/L to 43 33±1 21 U/L and from 0 91±0 03 nmol/L to 1 71±0 02 nmol/L, respectively, whereas the activity of SOD decreased from 33 48±1 15 U/ml to 16 93±1 11 U/ml (P<0 01). In hypoxic cardiomyocytes pretreated with 0 01-1 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±1 5 s and i increased by 107±13 2% during perfusion of hypoxic solution (P<0 001) at the end of 30 min. ET-1 (0 01-1 nmol/L) increased the frequency of i transient in cultured cardiomyocytes in a dose-dependent manner (P<0 01). The termination of i transient and the elevation of i caused by hypoxia were postponed by pretreatment with 0 01-1 nmol/L

关 键 词：内皮素 低氧 心肌细胞 细胞内钙 

分 类 号：R363[医药卫生—病理学]