不同浓度龙血竭总黄酮对大鼠MIRI心肌细胞PI3K/AKT通路的影响

Effects of different concentrations of sanguis draconis flavones on PI3K/AKT pathway in myocardial cells of rats with MIRI

目的观察不同浓度龙血竭总黄酮(SDF)对大鼠MIRI模型的治疗作用,探讨其对PI3K/AKT信号通路及凋亡蛋白的影响。方法56只SD雄性大鼠按随机数字表法分成Control组、Sham组、I/R组、龙血竭总黄酮低剂量组、中剂量组、高剂量组、大剂量组,每组8只。采用结扎冠状动脉左前降支法制备I/R损伤模型。采用酶联免疫吸附测定法检测大鼠血清乳酸脱氢酶(LDH)和肌酸激酶同工酶(CK-MB)含量;HE染色观察心肌病理学改变;TTC染色测定心肌梗死面积;Western Blot检测大鼠心肌PI3K、AKT、Caspase-3蛋白水平。结果①与I/R组相比较,SDF-L组、SDF-M组及SDF-H组、SDF-B组LDH和CK-MB表达水平降低(P<0.01)、心肌梗死面积减少(P<0.05)、PI3K、AKT蛋白含量升高(P<0.05)、Caspase-3蛋白含量减少(P<0.05)。②HE染色I/R组心肌细胞结构不完整,心肌细胞紊乱,细胞变性、坏死和炎症细胞浸润程度明显。不同浓度龙血竭总黄酮给药剂量组损伤较I/R组均减轻。结论龙血竭总黄酮预处理可通过激活PI3K/AKT信号通路,抑制细胞凋亡,对缺血再灌注损伤的心肌起到保护作用。

Objective To observe the therapeutic effects of sanguis draconis flavones(SDF)at different concentrations on myocardial ischemia-reperfusion injury(MIRI)in a rat model and explore its effects on the PI3K/AKT signaling pathway and apoptotic proteins.Methods Fifty-six male SD rats were randomly divided into 6 groups:control group,sham group,I/R group,SDF low-dose group,SDF medium-dose group,SDF high-dose group,and SDF big-dose group,with 8 rats in each group.I/R injury was induced by ligation of the left anterior descending coronary artery.Serum lactate dehydrogenase(LDH)and creatine kinase isoenzyme(CK-MB)concentrations were detected using enzyme-linked immunosorbent assay(ELISA);myocardial pathological changes were observed using HE staining;myocardial infarction size was measured using TTC staining;and protein levels of PI3K,AKT,and Caspase-3 were detected using Western Blot in rat myocardium.Results①Compared with the I/R group,the expression levels of LDH and CK-MB were decreased,myocardial infarction size was reduced(P<0.05),and the protein levels of PI3K and AKT were increased(P<0.05),while the protein levels of Caspase-3 were decreased(P<0.05)in the SDF low-dose group,SDF medium-dose group,SDF high-dose group,and SDF big-dose group.②HE staining showed that the myocardial cell structure in the I/R group was incomplete with disordered myocardial cells,obvious cell degeneration,necrosis,and infiltration of inflammatory cells.In contrast,the degree of injury was reduced in the sanguis draconis flavones groups with different concentrations compared with the I/R group.Conclusion Pre-treatment with sanguis draconis flavones at different concentrations can protect against myocardial ischemia-reperfusion injury by activating the PI3K/AKT signaling pathway and inhibiting cell apoptosis.