通络救脑注射液对大鼠脑微血管内皮细胞活性及其条件培养液影响的初步研究

Observing effects of TLJN injection on the rat cerebral microvascular endothelial cells and primary study of the feature of cellular conditioned medium

目的:观察通络救脑注射液对正常及拟缺血大鼠脑微血管内皮细胞的活性影响,并初步探讨细胞条件培养液内蛋白分泌的时效特征、奠定可溶性蛋白深入分析的技术基础。方法:通络救脑注射液作用于正常及拟缺血脑微血管内皮细胞之后,用MTS/PMS比色分析法测定细胞的活性,Bradford法测定细胞培养液总蛋白含量,比色分析法测定细胞培养液乳酸脱氢酶(LDH)漏出值,同步观察了5个时间点的细胞活性及条件培养液总蛋白量及LDH释放量。结果:通络救脑注射液能够提高拟缺血细胞的活性,且抑制LDH释放量;正常组分泌总蛋白量3h达到高峰,此时细胞活性最佳,LDH释放量亦少。拟缺血组分泌总蛋白量是6h达到高峰,此时LDH释放量最少,但细胞活性与3h比较有所下降。结论:通络救脑注射液对拟缺血细胞损伤具有保护作用;以3h至6h的细胞条件培养液做为收集目标是研究条件培养液的最佳时间段。

Objective： Through observing the active influence of tongluojiunao（TLJN） injection on the normal and ischemic rat cerebral microvascular endothelial cells（CMECs） and primary discussion on total protein secretive time - effect feature in cellular conditioned medium, to establish a technical basis for further study of solubility protein. Methods： After TLJN injection having reacted upon normal and ischemie cerebral microvascular endothelial cells（CMECs）, MTS/PMS colorimetric method, Bradford assay and eolorimetrie analysis antigenic were used respectively and isochronously to test cellular activity, total protein quantity of conditioned medium, and LDH releasing value of conditioned medium, observing them and analyzing the time - effect feature of cellular conditioned medium, Results ： TLJN injection can not only promote the activity of ischemic cells, but also reduce LDH releasing value, In the normal group, the peak time of total protein quantity is at 3rdh, at this time, cell activity is highest and LDH releasing value is least. In the ischemia group, the peak time of total protein quantity is at 6dda, simultaneously, LDH releasing value is least, but cell activity is a little lower than that of 3rdh. Conclusion： TLJN injection has protective action on ischamic cells. In this study, it is suggested that regarding conditioned medium from 3rdh to 6thh as collecting aim is the best period to study biological effects of solubility protein.