通络救脑注射液对脑微血管内皮细胞活性影响的特征

Effects of Tongluojiunao Injection on the Activity of Rat Cerebral Microvascular Endothelial Cells

目的:观察通络救脑注射液对培养的正常及缺血脑微血管内皮细胞的活性影响,揭示其通络作用的效应靶点与特征。方法:原代培养大鼠脑皮质微血管内皮细胞,传至第三代,分为正常及拟缺血组,采用培养基氧糖剥夺(OGD)法建立拟缺血模型。通过四甲基偶氮唑盐(MTT)比色分析法测定不同浓度的通络救脑注射液对正常及OGD内皮细胞的活性影响。结果:通络救脑注射液作用于正常脑微血管内皮细胞,与未加药组比较,小剂量药物抑制细胞活性趋势,大剂量促进细胞活性趋势,剂量总趋势呈反抛物线形;通络救脑注射液作用于OGD组脑微血管内皮细胞,小剂量范围促进内皮细胞的增殖活性,呈显著和极显著差异,而大剂量组则抑制细胞活性,剂量总趋势呈抛物线形。结论:通络救脑注射液对正常及缺血脑微血管内皮细胞具有双向调节作用,药物剂量与细胞增殖活性呈非线性关系。大剂量与小剂量可能是不同的作用机制,反映了中药复方药效的多维性。

Objective： To study the influence of Tongluojiunao（TLJN） injection on the normal cultivated rat cerebral microvascular endothelial cells（CMECs） and ischemic model ones respeetively, and to indicate its effect target and characteristics. Methods： Cerebral microvascular endothelial cells isolated from the male SD rats were primarily cultured and subcultured to the third generation for experiment and divided into normal and ischemic sets for study. The oxygen and glucose deprivation （OGD） method was adopted to construct ischemic model in vitro. The effects of TLJN injection of different concentration on the CMECs activity were determined by MTT[3- （4,5 - dimethylthiazol- yl） 2, 5 - diphenyltetrazolium bromide] colorimetric method. Results： On the normal cells group, the activity of cells was reduced by low dose of TLJN injection, but promoted by its large dose. It was then concluded that counter para- curve relationship existed between cell activity and drug dosage. On the other hand, Low dose of TLJN injection increased the activity of cells in the ischemic model group with significant deviation compared with negative control samples without TLJN injection, while large dose of TLJN injection reduced its activity with a para - curve relationship to the total dosage. Conclusion： TLJN injection had two- ways regulation on both the normal cerebral microvascular endothelial ceils and ischemic model ones, and there was non - linear relationship between drug dosage and cell generation activity. Low dosage and large dosage might have two different action mechanisms, which shows the multiple dimensional pharmacodynamic feature of the compound formula of traditional Chinese medicine （TCM）.