益心通脉颗粒基于PI3K/AKT通路对CHD大鼠心肌损伤及应激水平的研究

目的 探讨益心通脉颗粒基于PI3K/AKT通路对冠状动脉粥样硬化性心脏病(coronary atherosclerotic heart disease，CHD)大鼠心肌损伤及应激水平的研究。方法 将60只PF级Wistar健康大鼠按照随机数字法分为6组：A组（正常组）、B组（CHD组）、C组（格列吡嗪药物对照组）、D组（低剂量益心通脉颗粒组）、E组（中剂量益心通脉颗粒组）及F组（高剂量益心通脉颗粒组），每组10只，除A组外其余均建立CHD动物模型，建模成功后，C组大鼠给予0.5 mg/kg格列吡嗪溶于2 ml蒸馏水灌胃；D组、E组及F组大鼠均给予1.25 mg/kg、2.5 mg/kg及5 mg/kg的益心通脉颗粒溶于2 ml蒸馏水灌胃；其余组别大鼠灌胃2 ml生理盐水。试剂盒采用血清氧化应激指标；HE染色观察心肌组织形态及结构；TUNEL法检测心肌细胞凋亡率；免疫组化及免疫印迹分别检测心肌组织中PI3K/AKT阳性率及蛋白表达。结果 与A组相比，B组大鼠血清丙二醛（malondialdehyde，MDA）含量升高，NO、超氧化物歧化酶(super oxide dismutase，SOD)及谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）含量降低（P < 0.05），与B组相比，C组、D组、E组、F组大鼠血清MDA降低，其余指标升高（P < 0.05）；A组心肌组织结构清晰，纤维布局完整，B组大鼠心肌组织分布紊乱及纤维溶解坏死，C组、D组、E组及F组大鼠心肌结构改善；与A组相比，B组大鼠心肌细胞凋亡率升高（P < 0.05），C组、D组、E组及F组大鼠心肌细胞凋亡率均低于B组（P < 0.05）；B组大鼠心肌组织PI3K、AKT细胞阳性数量及蛋白水平均低于A组（P <0.05），与B组相比，C组、D组、E组及F组大鼠心肌组织PI3K、AKT细胞阳性数量及蛋白含量均升高（P < 0.05），且呈现浓度依赖性F组大鼠氧化应激水平、心肌细胞凋亡及PI3K、AKT细胞阳性数量均优于D组及E组，与C组相比无明显意义（P < 0.05）。结论 益心通脉颗粒能够通过降低CHD大鼠氧化应激水平，减少心肌组织病理损伤及心肌细胞凋亡，这可能与上调PI3K/AKT通路表达相关。

The study of Yixin Tongmai Granules on myocardial injury and stress level of rats with coronary heart disease based on PI3K/ AKT pathway

Objective To investigate the effects of Yixin Tongmai granules on myocardial injury and stress level of rats with coronary heart disease based on the PI3K/ AKT pathway. Methods 60 PF-class Wistar healthy rats were randomly divided into 6 groups including Group A (normal group), group B (CHD group), group C (glipizide drug control group), group D (low-dose Yixin Tongmai granule group), group E (medium-dose Yixin Tongmai granule group) ) and group F (high-dose Yixin Tongmai granule group), 10 rats in each group. Except for group A, all others had established coronary heart disease. After the model was successfully built, Rats in group C were given 0.5 mg/kg glipizide dissolved in 2 ml distilled water by gavage. Rats in group D, group E, and group F were given 1.25 mg/kg, 2.5 mg/kg and 5 mg/kg Yixintongmai granules dissolved in 2 ml distilled water for intragastric administration. The rats in the other groups were given 2 ml of normal saline by gavage. Results Compared to group A, the serum MDA content of group B was increased, while the contents of NO, SOD and GSH-Px were decreased (P< 0.05). Compared to group B, serum MDA in groups C, D, E and F was decreased, while other indexes were increased (P< 0.05). The myocardial tissue structure of group A was clear and the fiber layout was complete. The myocardial tissue distribution of group B was disordered and the fiber lytic necrosis was observed. The myocardial structure of groups C, D, E, and F was improved. Compared to group A, the apoptosis rate of myocardial cells in group B was increased (P< 0.05). The apoptosis rate of myocardial cells in groups C, D, E and F was lower than that of in group B (P< 0.05). The number of positively expressed cell and protein levels of PI3K and AKT in the myocardium of rats in group B were lower than those in group A (P<0.05). Compared with group B, the number of the positively expressed cell of PI3K and AKT and protein content in the myocardium of rats in groups C, D, E, and F were increased (P< 0.05).In addition, the levels of oxidative stress, myocardial apoptosis, and PI3K and AKT expression level in concentration-dependent group F were better than those in group D and E, but had no difference compared with group C (P> 0.05). Conclusion Yixin Tongmai Granules can reduce oxidative stress level, pathological damage of myocardial tissue and apoptosis of myocardial cells in rats with coronary heart disease, which may be related to the up-regulation of PI3K/ AKT pathway expression.