脑心通胶囊“心脑同治”的免疫和炎症标志物研究

Immune and inflammation markers of simultaneous treatment of heart and brain in Naoxintong Capsules

目的探讨脑心通胶囊“心脑同治”的免疫和炎症标志物。方法采用结扎左前降支冠状动脉(left anterior descending coronary artery ligation,LAD)模型为心肌缺血模型,大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)模型为脑缺血模型。雄性SD大鼠随机分为8组,包括用于心肌缺血研究的假手术组、LAD模型组、脑心通(110 mg/kg)组和卡托普利(10.125 mg/kg)组,以及用于脑缺血研究的假手术组、MCAO模型组、脑心通(220 mg/kg)组和银杏提取物(50 mg/kg)组,每组15只。药效学指标选择心电图、心肌酶谱、透射电镜等;生物信息学和蛋白质组学预测并筛选免疫炎症标志物;利用qRT-PCR和Western blotting验证关键备选标志物靶点的mRNA和蛋白相对表达量。结果脑心通胶囊不仅降低LAD大鼠的心梗死率、心肌酶活性(P<0.01),改善心肌损伤,还能降低LAD大鼠的脑海马组织损伤和脑水肿(P<0.05)。对于MCAO大鼠,脑心通胶囊显著降低脑梗死率、神经功能评分和脑组织含水率(P<0.05、0.01),同时减轻心肌组织和细胞损伤,降低心肌酶活性(P<0.01)。根据生物信息学和蛋白质组学结果,Toll受体信号通路的相关靶点是“心脑同治”的关键所在,有成为标志物的潜力。在心肌缺血和脑缺血2种状态下,α-干扰素(interferon-α,IFN-α)、Toll样受体4(Toll-like receptor 4,TLR4)、TLR7、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)共4个靶点的mRNA和蛋白相对表达量显著变化(P<0.05、0.01),表明LAD大鼠和MCAO大鼠的海马中出现炎症因子风暴和免疫反应瀑布。结论IFN-α、TLR4、TLR7、TNF-α能够作为海马中“心脑互损”“心脑同治”的免疫和炎症标志物。

Objective To explore the immune and inflammatory markers of simultaneous treatment of heart and brain in Naoxintong Capsules(脑心通胶囊).Methods The left anterior descending coronary artery ligation(LAD)model was used as the myocardial ischemia model,and the middle cerebral artery occlusion(MCAO)model was used as the cerebral ischemia model.SD male rats were divided into eight groups,including sham group,model group,Naoxintong(110 mg/kg)group and captopril(10.125 mg/kg)group used for myocardial ischemia research,as well as sham group,MCAO model group,Naoxintong(220 mg/kg)group and Ginkgo biloba extract(50 mg/kg)group used for cerebral ischemia research,with 15 rats in each group.Electrocardiogram,myocardial enzyme spectrum and transmission electron microscope were selected as pharmacodynamic indexes.Bioinformatics and proteomics were used to predict and screen immunoinflammatory markers.qRT-PCR and Western blotting were used to verify the mRNA and protein relative expression levels of key candidate marker targets.Results Naoxintong Capsules not only decreased the heart infarction rate and myocardial enzyme activity in LAD rats(P<0.01),improved myocardial injury,and decreased cerebral hippocampus injury and cerebral edema in LAD rats(P<0.05).In MCAO rats,Naoxintong Capsules significantly decreased cerebral infarction rate,neural function score and rate of water content in cerebral tissue(P<0.05,0.01),alleviated myocardial tissue and cell damage,and decreased myocardial enzyme activity(P<0.01).According to the results of bioinformatics and proteomics,the related targets of Toll receptor signaling pathway were the core for the simultaneous treatment of heart and brain and had the potential to become markers.The mRNA and protein relative expression levels of interferon-α(IFN-α),Toll-like receptor 4(TLR4),TLR7 and tumor necrosis factor-α(TNF-α)were significantly changed under two conditions of myocardial ischemia and cerebral ischemia(P<0.05,0.01),indicating that inflammatory cytokine storms and immune response cascades occurred in the hippocampus of LAD and MCAO rats.Conclusion IFN-α,TLR4,TLR7 and TNF-αcan be used as immune and inflammatory markers of heart-brain mutual damage and simultaneous treatment of heart and brain in hippocampus.