摘要
目的:观察葛根素对缺氧再给氧损伤心肌细胞的保护作用,从细胞水平揭示葛根素的作用。方法:①实验于2003-09/2004-02在锦州医学院科技楼完成。选用生后两三天的SD大鼠10只。②常规培养心肌细胞两三天后,为了减少血清中的成分对实验结果的影响,更换含0.4g/L的小牛血清的培养基。培养瓶内细胞用缺糖缺氧的溶液后,直接通入体积分数0.95氮气+体积分数0.05二氧化碳培养。在上述缺糖缺氧3h后,换用正常含糖含氧的Hanks液培养1和6h(缺氧再给氧组),在再给氧损伤之前,分别加入葛根素50,100及200mg/L(缺氧+50,100和200mg/L葛根素再给氧组)。正常组:加入相等数量的培养基,不加入任何药物。③将心肌死亡细胞及活细胞剪下分别称质量。计算死亡细胞的相对质量。以确定心肌细胞单层的坏死范围(心肌细胞活力)。④通过计数器计数给药前后心肌细胞搏动频率的变化。⑤采用全自动生化分析仪测定培养液上清乳酸脱氢酶含量。⑥计量资料差异比较采用方差齐性分析和t检验。结果:①心肌细胞的活力:缺氧+50和100及200mg/L葛根素再给氧组在再给氧1和6h后明显高于缺氧再给氧组犤(104.3±19.5)%,(83.4±18.2)%,(75.5±17.6)%,(190.2±36.7)%;(93.4±17.5)%,(71.8±16.1)%,(64.2±15.4)%,(183.4±32.8)%,P<0.01犦。②心肌细胞的搏动频率:正常组和3个给药组在再给氧1和6h后明显大于缺氧再给氧组(P<0.01)。③心肌细胞培养液上清乳酸氢清酶含量:正常组和3个给药组在再给氧1和6h后明显低于缺氧再给氧组(P<0.01)。结论:葛根素可提高损伤心肌细胞活力和搏动频率,降低心肌细胞培养液上清乳酸脱氢酶的含量,该种作用与再给氧时间关系不大。
AIM： To explore the protective effects of puerarin on hypoxia-reoxygenation-induced injury in cultured cardiac myocytes. METHODS： ① The experiment was performed in the Scientific Technological building of Jinzhou Medical College from September 2003 to February 2004. Ten Sprague-Dawley rats, 1-3 days old, were used. ②Mter the cardiac myocytes were regularly cultured for 2 or 3 days, the culture medium containing 0.4 g/L bovine serum was changed in order to reduce the effect of the components in serum on the results of experiment. The hypoxic conditions were simulated by exposure of cultured rat cardiomyocytes （2-3 days in vitro）, to an atmosphere mixture containing nitrogen gas （volume fraction was 0.95） and carbon dioxide （volume fraction was 0.05） in glucose-free medium for 3 hour, and then the cardiomyocytes were cultured in Hanks fluid containing normal glucose and normal oxygen for 1 and 6 hours （hypoxia/reoxygenation group）, before reoxygenation injury, puerarin of 50, 100 and 200 mg/L were given respectively （hypoxia/reoxygenation ＋puerarin 50, 100 and 200 mg/L groups）. In the normal group, the culture mediums of the same number were added without any drugs. ③ The dead and survived cardiomyoeytes were cut and weighed. The relative mass of the dead cells was calculated. The unilaminar myocardial necrosis range （myocardial activity） was identified. ④ The changes of the myocardial beat frequency before and after administration were counted. ⑤The content of lactate dehydrogenase in the supernatant of the culture medium was detected with automatic biochemistry analyzer. ⑥ The differences of the measurement data were compared with the homogeneity test of variances and t test. RESULTS： ① Myocardial activity： It was obviously higher in the hypoxia/reoxygenation ＋puerarin 50, 100 and 200 mg/L groups at 1 and 6 hours after hypoxia/reoxygenation than in the hypoxia/reoxygenation group [（104.3 ± 19.5）%, （83.4 ±18.2）%, （75.5 ±17.6）%, （190.2±36.7）% ; �
出处
《中国临床康复》
CSCD
北大核心
2005年第31期132-133,共2页
Chinese Journal of Clinical Rehabilitation
