通络救脑注射液对氧糖剥夺大鼠脑微血管内皮细胞分泌巨噬细胞炎性蛋白-1β的影响

Effect of TongluoJiunao injection on MIP-1β secretion of cerebral micro vascular endothelial cells in rats with oxygen-glucose deprivation

关键词:  脑微血管内皮细胞  氧糖剥夺  巨噬细胞炎性蛋白-1β  通络方药

Key words:  cerebral microvascular endothelial cells  oxygen-glucose deprivation  MIP-1β  Tongluo formula

摘要:

[目的]观察通络救脑注射液对氧糖剥夺损伤(OGD)的大鼠脑微血管内皮细胞条件培养液中巨噬细胞炎性蛋白-1β(MIP-1β)分泌情况及其受体——趋化因子受体5(CCR5)表达的影响。[方法]氧糖剥夺法建立内皮细胞OGD模型,酶联免疫银光法(ELISA)法检测脑微血管内皮细胞条件培养液中MIP-1β含量的变化,免疫组化法及Western blot法观察内皮细胞中MIP-1β及其受体CCR5的表达情况。[结果]OGD组的内皮细胞分泌大量MIP-1β至上清液,该组内皮细胞胞浆大量表达MIP-1β,同时CCR5的蛋白表达量亦随之上升,加入通络救脑注射液的OGD组,其上清液中MIP-1β分泌量显著性下降(P<0.01),MIP-1β及其受体CCR5在内皮细胞上的表达也有不同程度的下降。[结论]通络救脑注射液能够抑制OGD损伤时内皮细胞MIP-1β的分泌,减少MIP-1β和CCR5的表达,对内皮细胞缺氧损伤后趋化因子释放的抑制是其发挥解毒通络药效的途径之一。

Abstract:

[Objective] We addressed our study on the effect of Tongluo Jiunao injection (TLJN) on the MIP-1β secretion of cerebral mi-cro vascular endothelial cells and chemokine receptor 5 (CCR5) under the condition of oxygen-glucose deprivation in rats.[Methods] The oxygen-glucose deprivation was set up, and defined the third passage endothelial cells as four groups:normal endothelial cells (N),normal endothelial cells with TLJN (NT), oxygen-glucose deprivation cells (I), oxygen-glucose deprivation cells with TLJN (IT).The MIP-1βcontent of four supernatants was detected by ELISA and the MIP-1β and it’s receptor CCR5 expression were observed by immunohisto-chemistry and Western blot.[Results] Endothelial cells after deprivation of oxygen-glucose could secrete a lots of MIP-1β into the super-natant with increased expression of MIP-1β and CCCR5 in the injured cells.The content of MIP-1β in the medium of IT was significantlyreduced (P<0.01), the expression of MIP-1β and CCR5 was also decreased in different extent accordingly.[Conclusion] TLJN can signif-icantly inhibit the secretion of MIP-1β into the supernatant and the expression of MIP-1β and CCR5 when micro vascular endothelialcells was injured by oxygen-glucose deprivation.It was presumed that the TLJN’s effect of inhibiting the release of chemokine may be thepotential way of its pharmacodynamic action.