基于病证结合的清开灵/精制清开灵抗缺血性中风证治特点及其对神经血管单元的多靶点调控机制研究

Ischemic stroke a disease which TCM has definite treatment advantages. Numerous clinical studies have confirmed QKL is effective in the treatment of ischemic stroke, but its prevention and treatment characteristics and mechanisms of action needs to be further deepened. Neurovascular unit combines a synergy of variety cells in the brain. To carry out anti- cerebral ischemia study of traditional Chinese medicine using neurovascular unit as a carrier not only conform the therapeutic characteristics of TCM with multi-link and multi-target, but also helps to explain the underlying molecular mechanisms. This atudy intended to take two animal models of cerebral infarction as object, QKL and JZQKL as interventions to explore the syndrome characteristics and effecter mechanisms of QKL in treating different types of ischemic stroke from the dual perspective of disease and syndrome. We take neurovascular unit as a starting point to explain the temporal and spatial variation and the affect of and associated protein markers, and clear the multiple levels in the brain metabolic distribution of JZQKL with traditional Chinese medicine small molecule monoclonal antibody technology the Chinese small-molecule monoclonal antibody technology. We will clarify the material basis and mechanism of JZQKL through the analysis of the relationship of pharmacodynamic effects and metabolic effects of traditional Chinese medicine ingredients.; based on this small sample to carry out targeted clinical validation trials. Then we intend to carry out small samples of clinical validation trials on this basis.The study has a significant meaning on improving the clinical efficacy of ischemic stroke, clearing the main symptom and effects target of QKL in treatment and prevention of this disease and further promoting the development of cerebral protection Chinese medicine.

缺血性中风是中医确有疗效的优势病种之一。大量临床研究证实清开灵是治疗该病的有效药物，但对其防治特点及作用机制的认识尚需进一步深入。神经血管单元综合了脑内多种细胞的协同作用，以其为切入点开展中药抗脑缺血的研究，既符合中药多环节、多靶点的治疗特点，又有助于阐释相关分子机制。本课题拟以两种脑梗死动物模型为研究对象，以清开灵和精制清开灵为干预手段，从疾病与证候双重角度探讨清开灵治疗不同类型缺血性中风的证候特点和效应机制；以神经血管单元为重点，阐释清开灵对其时空变化特征和相关蛋白标记物的影响，并借助中药小分子单克隆抗体技术明确精制清开灵在脑内多层次的代谢分布特征，通过分析药效与效应成分代谢、分布的有机关联明确其物质基础及作用机制；并在此基础上有针对性的开展小样本临床验证性试验。该研究对明确清开灵治疗缺血性中风的证效、时效、量效特点，提高该病的临床疗效以及开发安全有效的脑保护中药均具有重要意义。

缺血性中风是中医确有疗效的优势病种之一。大量临床研究证实清开灵是治疗该病的有效药物，但对其防治特点及作用机制的认识尚需进一步深入。神经血管单元综合了脑内多种细胞的协同作用，本课题组以其为切入点开展中药抗脑缺血的研究，既符合中药多环节、多靶点的治疗特点，又有助于阐释相关分子机制。模型制备及证候辨识方面，完成了MCAO及BCCAO两种大鼠模型的证候辨识，观察了不同特点的脑缺血大鼠的证候演变，制备了24月龄的老年高血压大鼠BCCAO模型；时效量效特点方面，明确了药物的6小时治疗时间窗和最佳给药剂量（中剂量，30mg/kg）；作用机制方面，完成了QKL及JZQKL的网络药理学研究，借助分子生物学技术及抗体芯片技术等，发现了药物在抗凋亡、抗内质网应激、抗氧化应激及抗炎等多方面的多靶点调控机制。利用激光共聚焦三维重建技术及体外神经血管单元构建，发现了药物对急性期胶质细胞增生的抑制作用及神经血管单元功能完整性的保护作用。利用双光子显微镜技术，动态观察模型小鼠活体状态下毛细血管的结构，发现QKL显着减轻脑缺血后脑屏障功能障碍中微血管结构的破坏和荧光渗漏，发现模型小鼠脑血管“无复流”现象。该研究对明确清开灵治疗缺血性中风的证效、时效、量效特点，提高该病的临床疗效以及开发安全有效的脑保护中药均具有重要意义。