甘露醇预处理减轻H9c2细胞缺氧复氧损伤的作用

Reduction of anoxia/reoxygenation-induced injury by mannitol pretreatment

目的研究甘露醇对H9c2细胞缺氧复氧损伤的保护作用。方法常规培养大鼠H9c2心肌细胞株,分为正常组、模型组和实验组,正常组常规培养不加干预,模型组缺氧3 h,复氧12 h,实验组用20 mmol·L^(-1)甘露醇预处理1 h,随后进行缺氧复氧损伤。用流式细胞仪检测各组细胞存活率,用分光光度计检测细胞培养液中丙二醛浓度、乳酸脱氢酶释放量及肌酸激酶含量。结果与正常组相比,模型组早期凋亡率(12.76±1.66)%、晚期凋亡率(30.78±3.35)%和总死亡率(43.54±4.09)%均明显增高(均P<0.01),细胞培养液中丙二醛含量[(16.65±1.19)nmol·mL^(-1)]增高,乳酸脱氢酶[(181.30±11.87)U·mL^(-1)]和肌酸激酶[(33.91±2.28)U·mL^(-1)]的释放增加(均P<0.01)。与模型组相比,实验组细胞早期凋亡率(3.98±0.84)%、晚期凋亡率(7.89±0.53)%和总死亡率(11.86±1.10)%均明显降低(均P<0.01),细胞培养液中丙二醛含量[(11.90±0.81)U·mL^(-1),P<0.05]降低,乳酸脱氢酶[(97.28±3.12)U·mL^(-1),P<0.01]和肌酸激酶[(21.33±1.15)U·mL^(-1),P<0.01]的释放均明显减少。结论甘露醇可减轻心肌细胞缺氧复氧损伤,具有心肌细胞保护作用。

Objective To investigate the protective effect of mannitol against anoxia / reoxygenation- induced injury in H9c2 cells.Methods The H9c2 cells were divide into three groups： normal group,model group and test group. The normal group was cultured with normal medium without any intervention,the model group was cultured under anoxia with 95% N_2＋ 5% CO_2 for 3 hours fllowed by incubated in 95% O_2＋ 5% CO_2 for 12 hours in order to get anoxia / reoxygenation injury. The test group was pretreated with 20 mmol·L-1 mannitol for 1 hour before anoxia / reoxygenation injury. The cells＇ viability was analyzed by flow cytometry. The concentrations of creatine kinase（ CK- MB）,lactate dehydrogenase（ LDH）,malondialdehyde（ MDA） were determined using spectrophotometer. Results Compared with normal group,the cell＇s early apoptosis rate（ 12. 76 ± 1. 66） %,late apoptosis rate（ 30. 78 ± 3. 35） % and total death rate（ 43. 54 ± 4. 09%） in model group obviously increased（ all P〈 0. 01） accompanied with the elevation of MDA [（ 16. 65 ± 1. 19） nmol · mL-1],LDH [（ 181. 30 ± 11. 87）U·mL-1]and CK- MB[（ 33. 91 ± 2. 28） U·mL-1]in culture medium（ all P〈 0. 01）. Compared with model group,the cell＇s early apoptosis rate（ 3. 98 ± 0. 84） %,late apoptosis rate（ 7. 89 ± 0. 53） % and total death rate（ 11. 86 ± 1. 10） % obviously decreased（ all P〈 0. 01）in test group accompanied with the reductions of MDA [（ 11. 90 ± 0. 81） nmol · mL-1,P〈 0. 05 ],LDH[（ 97. 28 ± 3. 12） U·mL-1,P〈 0. 01]and CK- MB [（ 21. 33 ± 1. 15） U·mL-1,P〈 0. 01]in culture medium.Conclusion Mannitol protected the cardiomyocytes from anoxia / reoxygenation injury.