肌肽对大鼠脑片缺氧缺糖／再灌损伤的保护作用

摘要

目的：在离体脑片缺氧缺糖／再灌损伤模型上，评价肌肽对脑组织的保护作用。方法肌肽预处理后，用缺氧缺糖／再灌（oxygen glucose deprivation／reperfusion，OGD／RP）来制备大鼠离体脑片损伤模型。以2，3，5-三苯基氯化四氮唑（2，3，5-triphenyl tetrazolium chloride，TTC）染色法检测脑片活性；HPLC法检测海马脑片中ATP、ADP、AMP含量；荧光法检测脑组织活性氧（ reactive oxygen species，ROS）。结果与对照组相比，缺氧缺糖／再灌损伤可以明显损伤大鼠海马脑片，TTC染色颜色变浅，A490 nm明显下降， ATP和ADP含量明显降低，而AMP含量明显升高，ROS明显升高，差异均具有统计学意义（P＜0.01）。与模型组相比，缺氧缺糖／再灌损伤前预先加入1000、200、40μg／mL肌肽预处理15 min可显著抑制缺氧缺糖／再灌引起的损伤，TTC染色颜色加深，A490 nm明显升高，ATP、ADP、AMP含量升高，ROS含量降低，差异均具有统计学意义（ P＜0.01）。结论肌肽可减轻缺氧缺糖／再灌导致的损伤，其机制可能与其改善脑组织能量代谢，增强抗氧化能力有关。%Objective To investigate effect of carnosine on oxygen glucose deprivation/reperfusion ( OGD/RP) induced injury in rat brain slices. Methods Injury of brain slices was determined by TTC methods.The contents of ATP, ADP and AMP were determined by high performance liquid chromatography.Reactive Oxygen species ( ROS) were determined by fluorescence methods.Results Compared with control group, rat hippocampal slices were significantly damaged by OGD/RP, indicated by light color and decreased A490 nm value of TTC staining.Meanwhile the contents of ATP and ADP were significantly decreased, and the content of AMP and ROS were significantly increased, the difference between two group was significant ( P<0.01).Pre-incubation with Carnosine (1000, 200, 40 μg/mL) significantly inhibited the light color and decreased A490 nm value of TTC staining, increased the contents of ATP, ADP and AMP, and decreased the content of ROS, the difference between two group was significant ( P <0.01 ) . Conclusion Carnosine can protect rat hippocampal slices against injury induced by OGD/RP, which may relate to improve the energy metabolism and strengthen the ability of anti-oxidative stress.