通窍活血汤含药脑脊液对谷氨酸致PC12细胞损伤的保护作用

目的研究通窍活血汤含药脑脊液对谷氨酸致PC12细胞损伤的保护作用。方法大鼠灌胃给予通窍活血汤提取液制备含药脑脊液后,倒置显微镜下观察细胞形态,MTT法检测细胞活性,台盼蓝染色、LDH法观察细胞膜通透性,试剂盒法检测细胞内NO、MDA水平及SOD、ATP酶活性,DCFH-DA荧光探针染色检测细胞内活性氧(ROS)水平,Fura-2/AM染色检测细胞内游离Ca²⁺水平,流式细胞联合Annexin V-FITC/PI法分析细胞凋亡。结果与模型组比较,含药脑脊液组细胞形态显著改善,细胞活性、存活率显著升高(P<0.05,P<0.01),LDH活性,NO、ROS、MDA、游离Ca²⁺水平及细胞凋亡率显著降低(P<0.05,P<0.01),SOD、ATP酶活性显著升高(P<0.05,P<0.01)。结论通窍活血汤含药脑脊液对谷氨酸致PC12细胞损伤具有保护作用。

Protective effects of cerebrospinal fluid containing Tongqiao Huoxue Decoction on glutamic acid-induced PC12 cell injury

AIM To study the protective effects of cerebrospinal fluid containing Tongqiao Huoxue Decoction on glutamic acid-induced PC12 cell injury. METHODS Rats were given intragastric administration of Tongqiao Huoxue Decoction extract to prepare medicated cerebrospinal fluid, after which observation of cell morphology under inverted microscope, detection of cell viability by MTT assay, observation of cell membrane permeability by Trypan blue staining and LDH method, detection of intracellular NO, MDA levels and SOD, ATPase activities by kit assay, detection of intracellular levels of reactive oxygen species (ROS) and free Ca2+ by DCFH-DA fluorescence probe staining and Fura-2/ AM staining, respectively, analysis of cell apoptosis by flow cytometry combined with Annexin V-FITC/ PI assay were performed. RESULTS Compared with the model group, the medicated cerebrospinal fluid group demonstrated obviously improved cell morphology, significantly increased cell viability and survival rate (P<0.05, P<0.01), markedly decreased LDH activity, NO, ROS, MDA, free Ca2+ levels and cell apoptosis rate (P <0.05, P <0.01), significantly elevated SOD and ATPase activities (P <0.05, P <0.01). CONCLUSION The cerebrospinal fluid containing Tongqiao Huoxue Decoction has protective effects on glutamic acid-induced PC12 cell injury.