目的:观察脑出血大鼠心肌组织热休克蛋白70(HSP70)蛋白表达的变化及扶正护脑胶囊的干预作用。方法:动物随机分为假手术组、模型组、扶正护脑组和安宫牛黄组,分别于术后6 h、24 h、72 h 3个时相点取材,采用免疫组织化学SABC法测定心肌组...目的:观察脑出血大鼠心肌组织热休克蛋白70(HSP70)蛋白表达的变化及扶正护脑胶囊的干预作用。方法:动物随机分为假手术组、模型组、扶正护脑组和安宫牛黄组,分别于术后6 h、24 h、72 h 3个时相点取材,采用免疫组织化学SABC法测定心肌组织HSP70蛋白表达,并进行图像分析,观测其IOD值;并与血清CK-MB、cTnI含量作相关性分析。结果:HSP70阳性颗粒主要在心内膜下的心肌细胞和血管内皮细胞的胞浆中表达。模型组各时间点心肌组织HSP70蛋白表达均显著升高,其IOD值亦明显升高,扶正护脑组和安宫牛黄组均能明显增加HSP70蛋白的表达,升高其IOD值。IOD值与cTnI、CK-MB含量呈正相关。结论:HSP70对脑出血诱发的心肌损伤具有保护作用;扶正护脑胶囊通过提高HSP70的水平,减轻脑出血后心肌的损伤。展开更多
目的:观察通心脉方对大鼠离体心脏缺血再灌AKt及p38αMAPK(p38 a mitogen-activated protein kinase,MAPK)蛋白表达的影响,初步探讨通心脉方对心肌缺血再灌注损伤及细胞凋亡的作用机制。方法:采用Langendorff离体心脏灌流方法模拟缺血...目的:观察通心脉方对大鼠离体心脏缺血再灌AKt及p38αMAPK(p38 a mitogen-activated protein kinase,MAPK)蛋白表达的影响,初步探讨通心脉方对心肌缺血再灌注损伤及细胞凋亡的作用机制。方法:采用Langendorff离体心脏灌流方法模拟缺血再灌注模型;分光光度法观察心肌组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)、caspase-8及caspase-3含量的变化,酶联免疫(ELISA)法观察心肌组织中肿瘤坏死因子-α(TNF-α)含量的变化,免疫印迹(Western blot)法检测心肌组织p-p38αMAPK(phosphoryl-ated p38αmitogen activated protein kinase)、p38αMAPK、p-AKt(phosphorylated AKt)及AKt蛋白表达的变化。结果:与缺血再灌注组对比,正常灌流组、通心脉方大、中剂量组及卡维地洛组MDA、TNF-α、caspase-8及caspase-3含量显著降低,差别有统计学意义(P<0.01或P<0.05),SOD及CAT含量显著升高,差别有统计学意义(P<0.05或P<0.01);p38αMAPK及AKt在各组表达相对恒定,与缺血再灌注组相比,差别无统计学意义(P>0.05),p-p38αMAPK在与缺血再灌注组明显激活,在通心脉方大、中剂量组及卡维地洛组表达明显降低,差别有统计学意义(P<0.01);p-AKt在与缺血再灌注组激活,而在通心脉方大、中剂量组及卡维地洛组表达进一步增加(P<0.05)。结论:通心脉方能保护心肌缺血再灌注损伤和细胞凋亡,其作用机制可能与通过抑制脂质过氧化,下调p-p38αMAPK表达,减少TNF-α的形成,进而抑制caspase-8及caspase-3活性以及激活细胞增殖信号AKt有关。展开更多
Objective.To investigate the protective effects of melatonin(MT )on cardiomyocytes against the ox-idative injury induced by H 2 O 2 .Methods.Ventricular myocytes were isolated from neonatal rats and cultured for3～5da...Objective.To investigate the protective effects of melatonin(MT )on cardiomyocytes against the ox-idative injury induced by H 2 O 2 .Methods.Ventricular myocytes were isolated from neonatal rats and cultured for3～5days.Cells were divided into4groups:control group,H 2 O 2 (100μmol/L H 2 O 2 )group,MT group(100μmol/L MT )and MT preconditioning group(100μmol/L MT+100μmol/L H 2 O 2 ).The fluorescent probe,DCFH?DA,was used to detect intracellular levels of reactive oxygen species(ROS)and another fluorescent probe,Fluo-3?AM,was used to detect[Ca 2+ ] i by using a laser scanning confocal microscopy(LSCM).The malondi-aldehyde(MDA)content in cardiomyocytes was determined by measuring thiobarbituric acid?reactive sub-stances to monitor lipid peroxidation.The activity of cytoplasmic enzyme lactate dehydrogenase(LDH)that was released into the culture media was assayed to indicate alternation in the integrity of the cellular membrane.Trypan blue exclusion was used to detect the cell viability.Results.Compared with the control group,intracellular ROS,[Ca 2+ ] i ,MDA content ,LDH leakage and cell death were significantly elevated when cells were treated by100μmol/L H 2 O 2 for60minutes(P<0.01).However,those changes were significantly attenuated in MT preconditioning group.Conclusion.MT has very good antioxidant effect and can protect the cardiomyocytes against H 2 O 2 -in-duced injury.展开更多
文摘目的:观察脑出血大鼠心肌组织热休克蛋白70(HSP70)蛋白表达的变化及扶正护脑胶囊的干预作用。方法:动物随机分为假手术组、模型组、扶正护脑组和安宫牛黄组,分别于术后6 h、24 h、72 h 3个时相点取材,采用免疫组织化学SABC法测定心肌组织HSP70蛋白表达,并进行图像分析,观测其IOD值;并与血清CK-MB、cTnI含量作相关性分析。结果:HSP70阳性颗粒主要在心内膜下的心肌细胞和血管内皮细胞的胞浆中表达。模型组各时间点心肌组织HSP70蛋白表达均显著升高,其IOD值亦明显升高,扶正护脑组和安宫牛黄组均能明显增加HSP70蛋白的表达,升高其IOD值。IOD值与cTnI、CK-MB含量呈正相关。结论:HSP70对脑出血诱发的心肌损伤具有保护作用;扶正护脑胶囊通过提高HSP70的水平,减轻脑出血后心肌的损伤。
文摘目的:观察通心脉方对大鼠离体心脏缺血再灌AKt及p38αMAPK(p38 a mitogen-activated protein kinase,MAPK)蛋白表达的影响,初步探讨通心脉方对心肌缺血再灌注损伤及细胞凋亡的作用机制。方法:采用Langendorff离体心脏灌流方法模拟缺血再灌注模型;分光光度法观察心肌组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)、caspase-8及caspase-3含量的变化,酶联免疫(ELISA)法观察心肌组织中肿瘤坏死因子-α(TNF-α)含量的变化,免疫印迹(Western blot)法检测心肌组织p-p38αMAPK(phosphoryl-ated p38αmitogen activated protein kinase)、p38αMAPK、p-AKt(phosphorylated AKt)及AKt蛋白表达的变化。结果:与缺血再灌注组对比,正常灌流组、通心脉方大、中剂量组及卡维地洛组MDA、TNF-α、caspase-8及caspase-3含量显著降低,差别有统计学意义(P<0.01或P<0.05),SOD及CAT含量显著升高,差别有统计学意义(P<0.05或P<0.01);p38αMAPK及AKt在各组表达相对恒定,与缺血再灌注组相比,差别无统计学意义(P>0.05),p-p38αMAPK在与缺血再灌注组明显激活,在通心脉方大、中剂量组及卡维地洛组表达明显降低,差别有统计学意义(P<0.01);p-AKt在与缺血再灌注组激活,而在通心脉方大、中剂量组及卡维地洛组表达进一步增加(P<0.05)。结论:通心脉方能保护心肌缺血再灌注损伤和细胞凋亡,其作用机制可能与通过抑制脂质过氧化,下调p-p38αMAPK表达,减少TNF-α的形成,进而抑制caspase-8及caspase-3活性以及激活细胞增殖信号AKt有关。
文摘Objective.To investigate the protective effects of melatonin(MT )on cardiomyocytes against the ox-idative injury induced by H 2 O 2 .Methods.Ventricular myocytes were isolated from neonatal rats and cultured for3～5days.Cells were divided into4groups:control group,H 2 O 2 (100μmol/L H 2 O 2 )group,MT group(100μmol/L MT )and MT preconditioning group(100μmol/L MT+100μmol/L H 2 O 2 ).The fluorescent probe,DCFH?DA,was used to detect intracellular levels of reactive oxygen species(ROS)and another fluorescent probe,Fluo-3?AM,was used to detect[Ca 2+ ] i by using a laser scanning confocal microscopy(LSCM).The malondi-aldehyde(MDA)content in cardiomyocytes was determined by measuring thiobarbituric acid?reactive sub-stances to monitor lipid peroxidation.The activity of cytoplasmic enzyme lactate dehydrogenase(LDH)that was released into the culture media was assayed to indicate alternation in the integrity of the cellular membrane.Trypan blue exclusion was used to detect the cell viability.Results.Compared with the control group,intracellular ROS,[Ca 2+ ] i ,MDA content ,LDH leakage and cell death were significantly elevated when cells were treated by100μmol/L H 2 O 2 for60minutes(P<0.01).However,those changes were significantly attenuated in MT preconditioning group.Conclusion.MT has very good antioxidant effect and can protect the cardiomyocytes against H 2 O 2 -in-duced injury.
