基于PI3K/Akt信号通路异丹叶大黄素抗大鼠脑缺血再灌注损伤的神经元凋亡机制研究  

Study on Mechanism of Isosalvia Leaf Emodin Resisting Neuron Apoptosis Induced by Cerebral Ischemia reperfusion Injury in Rats Based on PI3K/Akt Signal Pathway

作　　者：孙家贺 安文灿 SUN Jiahe;AN Wencan(Sishui County Peoples Hospital,Jining 273200,Shandong,China;Qingdao Hospital of Peking University Peoples Hospital,Qingdao 266000,Shandong,China)

机构地区：[1]泗水县人民医院,山东济宁273200 [2]北京大学人民医院青岛医院,山东青岛266000

出　　处：《实用中医内科杂志》2025年第4期139-142,143,I0029,I0030,共7页Journal of Practical Traditional Chinese Internal Medicine

基　　金：青岛大学医疗集团科研专项基金资助项目(YLJT20202035)。

摘　　要：目的探讨异丹叶大黄素(Isorhapontigenin,ISO)调节PI3K/AKT信号通路,进而减轻缺血再灌注小鼠脑缺血半暗带神经元损伤的作用及机制。方法随机选择90只SD雄性大鼠,分成5组:(1)假手术组(n=20);(2)缺血/再灌注组即模型组(n=20);(3)ISO处理组(n=30,分成低剂量25 mg/kg、中剂量50 mg/kg、高剂量组100 mg/kg);(4)渥曼青霉素(PI3K抑制剂)组(n=10);(5)ISO(100 mg/kg)+渥曼青霉素(0.6 mg/kg)组(n=10)。假手术组给予手术但未穿线,模型组采用Longa线栓法建立脑缺血再灌注损伤,异丹叶大黄素(ISO)处理组再灌注后2 h腹腔内注射ISO,渥曼青霉素组在缺血前半小时脑室内注入渥曼青霉素(0.6 mg/kg)。参考Z-Longa5评分标准评估神经功能缺损情况,TTC染色法测定脑梗死率,TUNEL染色观察神经元凋亡情况,ELISA法检测髓过氧化物酶(MPO)含量,蛋白标记法(WB)检测磷酸化Akt、PI3K蛋白表达水平,定量qPCR检测抗凋亡因子Bcl-2、促凋亡因子Casepase-3、Bax基因表达量。结果本研究中ISO中、高剂量组大鼠神经功能损伤程度、脑梗死率、血清MPO水平均显著低于模型组,但添加PI3k抑制剂(渥曼青霉素)的大鼠,其神经功能缺损程度、脑梗死率及MPO水平均显著升高。TUNEL检测显示异丹叶大黄素(ISO)组较模型组可以有效降低脑细胞凋亡率。模型组中磷酸化AKT、PI3K蛋白表达水平明显下降。与模型组相比,异丹叶大黄素组AKT、PI3K蛋白表达明显升高。通过定量qPCR检测凋亡因子提示异丹叶大黄素有效提高抗凋亡因子Bcl-2基因表达量,降低促凋亡因子Casepase-3、Bax基因表达量。结论异丹叶大黄素(ISO)可能通过激活PI3K/AKT信号通路,调控氧化应激、抑制细胞凋亡,进而减轻脑缺血再灌注对脑细胞造成的损伤程度。Objective To verify the role and mechanism of Isosalvia leaf emodin(ISO)in alleviating neuronal damage in the ischemic penumbra of mice by regulating PI3K/AKT signaling pathway.Methods Ninety male SD rats were randomly selected and divided into 5 groups:sham surgery group(n=20),ischemia/reperfusion group(model group)(n=20),ISO treatment groups ISO low dose group(25 mg/kg,n=10),ISO medium dose group(50 mg/kg,n=10),and ISO high dose group(100mg/kg,n=10),Warmannin penicillin(PI3K inhibitor)group(n=10),ISO(100 mg/kg)+Warmannin penicillin(0.6 mg/kg)group(n=10).The sham surgery group was given surgery without threading.The model group used thread embolism method to establish cerebral ischemia-reperfusion injury.ISO treatment groups were given intraperitoneal injection of ISO 2 hours after reperfusion,and the Warmannin group was given intraventricular injection of Warmannin(0.6 mg/kg)half an hour before ischemia.The Z-Longa5 scoring standard was used to assess the neurological function defect.TTC staining was used to determine the cerebral infarction rate,TUNEL staining was used to observe the neuronal apoptosis.ELISA was used to detect the content of myeloperoxidase(MPO).Western blot(WB)was used to detect the expressions of phosphorylated Akt and PI3K proteins,and quantitative qPCR was used to detect the expressions of anti apoptotic factor Bcl-2,pro apoptotic factor Casepase-3 and Bax genes.Results The degree of neurological damage,cerebral infarction rate and serum MPO levels in the ISO medium and high-dose groups were significantly lower than those in the model group.However,the rats added with PI3k inhibitors(Warmannin penicillin)showed a significant increase in the degree of neurological damage,cerebral infarction rate and MPO levels.TUNEL test showed that ISO group could effectively reduce the rate of brain cell apoptosis compared with the model group.The expression levels of phosphorylated Akt and PI3K proteins significantly decreased in the model group.Compared with those of the model group,the expressions of A

关 键 词：异丹叶大黄素 PI3K/AKT 缺血再灌注损伤 氧化应激 神经元凋亡 

分 类 号：R285.5[医药卫生—中药学]