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目的检测2型糖尿病大鼠血液中一氧化氮(NO)、肌酸激酶(CK)、超氧化物歧化酶(SOD)及微量丙二醛(MDA)的变化情况和心肌组织的凋亡情况,探讨叶酸和维生素B_(12)干预对大鼠心肌的保护作用。方法六周龄SD大鼠60只,糖尿病造模成功后,随机分为对照组(A组)、模型组(B组)、叶酸和维生素B_(12)干预组(C组)。12周后心脏取血,检测各组NO、CK、SOD及MDA的水平;取大鼠心肌组织,Tunel法观察心肌组织的细胞凋亡情况;免疫组化法检测Caspase-3、Caspase-8的表达,并用计算机图像分析系统测平均灰密度值。结果 12周后心脏血液中,B组NO、SOD水平较A组明显下降,C组较B组明显增高(P<0.01);B组CK、MDA水平较A组明显增高,C组较B组明显下降(P<0.01);心肌组织中,三组的凋亡指数分别为(2.6357±0.5214)、(16.2859±0.6983)、(4.0410±0.3121);Caspase-3各组分别为(79.2471±2.3547)、(97.1530±4.9522)、(84.1549±3.0198);Caspase-8各组分别为(70.2596±3.2118)、(98.9322±5.0583)、(78.3299±3.5680)。B组与C组细胞凋亡指数、Caspase-3、Caspase-8的表达均高于A组(P<0.01),C组较B组明显下降(P<0.01)。结论 2型糖尿病大鼠血液中NO、SOD、CK、MDA的变化参与心肌细胞的凋亡过程,而叶酸和维生素B_(12)的干预可以有效缓解心肌损伤。
Objective To investigate the changes of blood nitric oxide (NO), creatine kinase (CK), superoxide dismutase (SOD) and malondialdehyde (MDA) in blood and the apoptosis of myocardium in type 2 diabetic rats, To explore the protective effect of folic acid and vitamin B_ (12) on myocardium in rats. Methods Sixty - six SD rats were randomly divided into control group (group A), model group (group B), folic acid and vitamin B - 12 intervention group (group C) after successful diabetes mellitus. Twelve weeks later, the heart blood was taken and the levels of NO, CK, SOD and MDA in each group were measured. The myocardial apoptosis was observed by Tunel method. The expressions of Caspase-3 and Caspase-8 were detected by immunohistochemistry Expression, and the use of computer image analysis system to measure the average gray value. Results After 12 weeks, the level of NO and SOD in group B was significantly lower than that in group A, but significantly higher in group C than that in group B (P <0.01). The levels of CK and MDA in group B were significantly higher than those in group A (P <0.01). The apoptotic indexes in the three groups were (2.6357 ± 0.5214), (16.2859 ± 0.6983) and (4.0410 ± 0.3121), respectively; the levels of Caspase-3 in each group were (79.2471 ± 2.3547) , (97.1530 ± 4.9522) and (84.1549 ± 3.0198), respectively. The levels of Caspase-8 were (70.2596 ± 3.2118) and (98.9322 ± 5.0583) and (78.3299 ± 3.5680) respectively. The apoptosis index, the expression of Caspase-3 and Caspase-8 in group B and group C were higher than those in group A (P <0.01), but decreased in group C compared with group B (P <0.01). Conclusion The changes of NO, SOD, CK and MDA in the blood of type 2 diabetic rats are involved in the process of cardiomyocyte apoptosis. However, the intervention of folic acid and vitamin B 12 can effectively relieve myocardial injury.