黄连解毒汤对PC12细胞和小鼠全脑缺血损伤的保护作用

Protective Effects of Huanglian-Jian-Du-Tang on Ischemic Injury in PC12 Cells and Global Cerebral Ischemia in Mice

[目的]观察黄连解毒汤(huanglian-jian-du-tang,HJDT)对PC12细胞和小鼠全脑缺血损伤的保护作用。[方法]建立体外糖氧剥夺(oxygen-glucose deprivation,OGD)和谷氨酸诱导的PC12细胞损伤模型,分别给予不同浓度的HJDT,以四甲基偶氮唑盐(MTT)和乳酸脱氢酶(LDH)法观察HJDT对细胞损伤的保护作用。采用双侧颈总动脉阻断30min建立C57BL/6小鼠全脑缺血模型,术前7d开始灌服HJDT(1、2、4、8 g.kg-)1至术后3d,采用新奇事物探究实验检测小鼠学习记忆能力,采用Nissl染色检测海马神经元存活的情况。[结果]OGD 4 h复灌2 h后,HJDT可显著增加PC12细胞活性,显著减少LDH释放(P<0.01);谷氨酸处理24 h后,HJDT能显著增加PC12细胞活性,减少LDH释放(P<0.05)。HJDT(4 g.kg-)1能显著提高全脑缺血小鼠对新物体的分辨比,(2、4 g.kg-)1可显著提高小鼠海马CA1区的神经元密度。[结论]HJDT对PC12细胞和小鼠全脑缺血损伤具有保护作用。

[Objective] To investigate the protective effects of Huanglian-Jian-Du-Tang（HJDT） on ischemic injury induced by oxygen-glucose deprivation（OGD） and glutamate in PC12 cells and on transient global cerebral ischemia in mice.[Methods] The PC12 cell injury was induced by OGD or glutamate in vitro.Different concentrations of HJDT were added into the PC12 cells.The effects of HJDT on cellular damage were respectively detected by MTT and LDH methods.Transient global cerebral ischemia in C57BL/6 mice was induced by occlusion of the bilateral common carotid arteries for 30 min.HJDT（1,2,4,8 g·kg-1）was orally administered to mice from day 7 before ischemia to day 3 after ischemia.Novel object recognition test was used to detect the learning and memory ability of mice.The survival neuron density was detected with Nissl staining.[Results] HJDT increased the cell viability ratio and reduced the LDH leakage ratio induced by OGD for 4 h followed by 2 h reoxygenation（P0.01）.HJDT increased the cell viability ratio and reduced the LDH leakage ratio induced by glutamate for 24 h（P〈0.05）.HJDT（4 g·kg-1） significantly increased the novelty object discrimination ratio and HJDT（2,4 g·kg-1） significantly increased the survival neuronal density in hippocampal CA1 after ischemia in mice.[Conclusion] Our results indicate that HJDT has protective effects on ischemic injury in PC12 cells in vitro and on transient global cerebral ischemia in mice in vivo.