丹参对ox-LDL孵育的乳鼠心肌细胞缺氧-复氧所致细胞凋亡的保护作用

Protective Effect of Salvia Miltiorrhiza on Apoptosis of ox-LDL Cultured Myocardial Cells Induced by Anoxia/reoxygenation in Neonate Rat

目的:观察丹参对ox-LDL孵育的乳鼠心肌细胞缺氧-复氧损伤的保护作用,并探讨其可能机制。方法:采用原代培养乳鼠的心肌细胞,ox-LDL孵育24小时,再采用95%N2+5%CO2模拟缺氧培养3小时后,更换为正常DMEM培养液培养9小时的方法建立心肌细胞缺氧/复氧模型。乳鼠心肌细胞随机分为6组:正常对照组(Control group);ox-LDL组(ox-LDL group);ox-LDL缺氧-复氧组(ox-LDL+A/R group);丹参保护组,加丹参低剂量(SM L/ox-LDL+A/R group)、中剂量(SM M/ox-LDL+A/Rgroup)、高剂量(SM H/ox-LDL+A/R group)组。利用MTT比色法检测心肌细胞存活率,Hoechst33342染色检测凋亡率,采用Western blot分别观察Cytc、Bax、bcl-2、caspase-3的蛋白表达。结果:通过MTT比色法检测,加入丹参高剂量组对细胞保护最有效。与Control组比较,ox-LDL组:细胞核有固缩和断裂,Cytc、Bax和caspase-3的表达升高,细胞存活率和bcl-2表达降低;与ox-LDL组比较,ox-LDL+A/R组:细胞核固缩和断裂加重,Cytc、Bax和caspase-3的表达进一步增加,细胞存活率和bcl-2表达则显著降低;SM/ox-LDL+A/R组:缺氧-复氧诱导的上述变化显著减轻。结论:丹参对ox-LDL孵育乳鼠心肌细胞的缺氧-复氧损伤具有保护作用,其机制与抑制氧化应激所致的细胞凋亡有关。

Objective:To observe protective effect of Salvia on ox-LDL cultured myocardial cells injured by anoxia/reoxygenation in neonate rat.Methods:The rat myocardial atherosclerosis cellular models were prepared using the primary cultured myocardial cells incubated with ox-LDL;the myocardial models of anoxia/reoxygenation:cells first were maintained in a humidified 37℃,95%N2-5%CO2 incubator for 3 hours,then replaced culture fluid with DMEM and cultured in normal condition for 9 hours.Cells of neonate rat were divi...