基于醛糖还原酶介导多元醇通路的丹红注射液抗大鼠脑缺血再灌注损伤的作用机制研究

Cerebral Ischemia Reperfusion Injury is severely harmful to the health of mankind. It has not yet made a breakthrough because the pathogenesis of the disease is complex. However, Our previous studies found that the key factor AR of polyol pathway was activated after cerebral ischemia reperfusion in rats. Moreover, Danhong injection (DHI) can inhibit the activity of AR, and reduce the cerebral ischemia reperfusion injury, which suggesting that AR may be a new target of DHI against cerebral ischemia reperfusion injury. At present, the prevention and control of this disease has achieved good clinical effect of combined traditional Chinese and Western Medicine. However, it has not been reported about the mechanism of Danhong injection (DHI) against the ischemic brain injury based on Aldose reductase (AR). Therefore, further study on the role of AR in this pathological process is of great significance to the discovery of new targets of the disease. In this study, the drug of the Danhong injection, which is the Traditional Chinese Compound Prescription, will be used as against cerebral ischemia reperfusion injury. We will make the Neuron-2 a cells into OGD model and make the SD rats into MCAO model, and then these methods such as Immunohistochemistry, Western bolt, Cerebral microdialysis and so on, which will be used in this study. And these scientific problems will be researched such as how to play the role of the activity and expression of AR in the brain damage, and how the relationship between polyol pathway and DHI. And these problems will be explained by the tissue, cellular and molecular level in our Group. Our ultimate objectives are to find out the role of AR and the protective mechanism of DHI in cerebral ischemia reperfusion injury. This study has important significance to find a drug target, to improve the clinical effect, and to provide experimental data and theoretical basis on early prevention and treatment in acute cerebral vascular disease.

脑缺血再灌注损伤严重危害人类健康，由于该病发病机制复杂，至今尚未取得突破性进展。申请人前期研究发现，大鼠脑缺血再灌注后多元醇通路关键因子醛糖还原酶（AR）激活，丹红注射液（Danhong Injection，DHI）能抑制AR活性减轻脑缺血再灌注损伤，提示AR可能是治疗该疾病的新靶点。目前以AR为切入点研究该病的作用机制还未见报道。因此，深入研究AR在此病理过程所扮演角色，对该疾病新靶点的发现具有重要意义。本项目以中药方剂DHI为工具药，制作体外Neuron-2α细胞OGD模型和体内SD大鼠MCAO模型，采用免疫组化、Western bolt、脑微透析等技术方法，在组织、细胞、分子水平上开展AR的病理表征及AR依赖的多元醇通路与DHI相关性研究，进一步揭示DHI抗脑缺血再灌注损伤作用机制，对药物靶点的发现，提高临床疗效以及急性期脑血管疾病的早期预防和治疗策略提供实验研究基础和理论依据。

脑缺血再灌注损伤机制涉及多种复杂的病理过程，其中脑组织炎症和神经元损伤是主要特点，最近研究表明，针对脑缺血再灌注损伤后炎症机制的关键环节加以干预可能成为治疗脑缺血再灌注损伤的有效途径。最近研究发现，多元醇通路的关键限速酶AR是多种炎症性疾病的重要节点。丹红注射液是临床用于治疗冠心病、心绞痛、心肌梗塞，缺血性脑病、脑血栓等有效制剂，现已发现丹红注射液对抗脑缺血损伤具有多种保护作用。然而AR是否参与脑组织炎性损伤及在该病理过程中扮演的角色以及DHI抗脑缺血再灌注损伤与多元醇通路关键因子AR的相关性研究还未见报道。.本项目从丹红注射液临床制剂有效性出发，通过构建体内、外脑缺血模型，结合采用先进的技术手段分别从细胞和整体动物水平研究了DHI抗脑缺血再灌注损伤与多元醇通路关键因子AR的相关性，揭示了多元醇通路关键因子AR在该疾病过程的作用以及HI通过调节氧化应激水平及AR介导的多元醇通路，进而降低缺血引起的炎症因子表达和细胞凋亡是 DHI 抗脑缺血再灌注损伤的分子机制。本研究进一步明确了多元醇通路关键因子AR在该疾病过程的分布和作用以及DHI预治疗脑缺血再灌注损伤与多元醇通路关键因子AR相关性，不仅为中药DHI抗脑缺血再灌注损伤治疗靶点的研究提供新途径，对进一步阐明该复方制剂中神经保护作用的药效物质基础，提高临床疗效，为临床用药提供理论基础和实验依据，同时也对相关神经退行性等疾病新的治疗靶点提供了新思路和新方法。