丹参川芎嗪注射液抗脑缺血缺氧损伤的药效物质基础研究

目的从注射液、组分、成分3个层面,探究丹参川芎嗪注射液抗脑缺血缺氧损伤作用及其药效物质基础。方法首先采用SH-SY5Y神经细胞构建氧糖剥夺/复氧(oxygen-glucosedeprivation/reoxygenation,OGD/R)损伤模型,以细胞活力为指标,考察丹参川芎嗪注射液对神经细胞的保护作用。其次,采用制备液相色谱等技术制备丹参川芎嗪注射液标准组分,然后在SH-SY5Y神经细胞OGD/R模型上,系统筛选丹参川芎嗪注射液组分和成分中抗脑缺血缺氧损伤的药效物质,最后进一步通过LC-MS对有效组分进行分析,并对有效成分进行归属,初步明确丹参川芎嗪注射液抗脑缺血缺氧损伤的药效物质基础。结果与模型组相比,丹参川芎嗪注射液在2.5%~15%的稀释浓度下,具有显著的抗SH-SY5Y细胞OGD/R损伤的作用;组分I、J、L、M、O、P、Q以及丹酚酸C、丹酚酸A、丹参素和盐酸川芎嗪均具有显著的抗SH-SY5Y细胞OGD/R损伤的作用。LC-MS分析结果表明,组分O、P中均含有丹酚酸C,组分O中含有丹酚酸A、盐酸川芎嗪,组分P中含有丹参素。结论丹参川芎嗪注射液具有显著的抗脑缺血缺氧损伤作用,而丹酚酸C、丹酚酸A、丹参素和盐酸川芎嗪可能是其抗脑缺血缺氧损伤的主要药效物质。

Study of the Effective Constituents of Danshen Chuanxiongqin Injection Against Hypoxic-Ischemic Brain Injury

OBJECTIVE To explore the effective constituents of Danshen Chuanxiongqin injection against hypoxic-ischemic brain injury from the three levels of injection, components and ingredients. METHODS Firstly, oxygen-glucose deprivation/reoxygenation(OGD/R) model was constructed in SH-SY5Y neuroblastoma cells. The protective effect of Danshen Chuanxiongqin injection on this ODG/R model was then detected by cell viability. Secondly, standard components of Danshen Chuanxiongqin injection were prepared by preparative liquid chromatography. The effective components and constituents of Danshen Chuanxiongqin injection were identified by the OGD/R model in SH-SY5Y cells. Finally, the active components and ingredients were analyzed by LC-MS, aiming to identify the effective constituents of Danshen Chuanxiongqin injection against hypoxic-ischemic brain injury. RESULTS Compared with the model group, Danshen Chuanxiongqin injection had a markedly protective effect on OGD/R injured SH-SY5Y cells in the dose range of 2.5%-15%. Further screening found that the components I, J, L, M, O, P, Q and salvianolic acid C, salvianolic acid A, Danshensu and ligustrazine hydrochloride could significantly alleviate OGD/R-induced injury in SH-SY5Y cells. The results of LC-MS analysis indicated that the components O and P contained salvianolic acid C, the component O contained salvianolic acid A and ligustrazine hydrochloride, and the component P contained Danshensu. CONCLUSION Danshen Chuanxiongqin injection has a markedly protective effect on hypoxic-ischemic brain injury, and salvianolic acid C, salvianolic acid A, Danshensu and ligustrazine hydrochloride may be the effective constituents of Danshen Chuanxiongqin injection against hypoxic-ischemic brain injury.