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  

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

作　　者：翟燕 汪宁[1,2] 章亚兵 王艳 ZHAI Yan;WANG Ning;ZHANG Ya-bing;WANG Yan(Anhui Provincial Key Laboratory for Chinese Medicine Compound,Ministry of Education Key Laboratory for Xin’ an Medicine,Anhui University of Chinese Medicine,Hefei 230012,China;Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine,Anhui Provincial Academy of Chinese Medicine,Hefei 230012,China)

机构地区：[1]安徽中医药大学中药复方安徽省重点实验室新安医学教育部重点实验室,安徽合肥230012 [2]安徽省中医药科学院中药药效与安全性评价研究所,安徽合肥230012

出　　处：《中成药》2019年第3期526-532,共7页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金项目(81773933;81374005);安徽省高校学科(专业)拔尖人才学术资助目(gxbjZD15);安徽省高校优秀中青年骨干人才国内外访学研修重点项目(gxfxZD2016117)

摘　　要：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Ca^(2+)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Ca^(2+)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。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.

关 键 词：通窍活血汤 脑脊液 PC12细胞 谷氨酸 

分 类 号：R285.5[医药卫生—中药学]