参附黄配方对脓毒症小鼠大脑皮质氧化损伤的影响

Effects of Shenfuhuang Formula(参附黄配方)on Oxidative Damage of Cerebral Cortex in Septic Sepsis Model Mice

目的 探讨参附黄配方对脓毒症神经元损伤的影响及作用机制。方法 C57BL/6J小鼠随机分为假手术组、脓毒症组和参附黄组,每组26只。假手术组小鼠麻醉后仅游离盲肠,不结扎及穿刺。脓毒症组和参附黄组采取盲肠结扎穿孔术(CLP)建立脓毒症模型。参附黄组造模后4 h予参附黄组方2.5 g/kg灌胃,每隔12 h进行下一次灌胃,每日灌胃2次,每次0.5 ml;脓毒症组和假手术组每次灌服纯净水0.5 ml,灌胃时间同参附黄组,各组均灌胃3 d。造模72 h后采用尼氏染色观察大脑前额叶皮层神经元数量的变化,采用二氢乙啶染色检测大脑皮质组织中活性氧含量,采用8-羟基脱氧鸟苷(8-OHdG)免疫荧光染色评价大脑皮质氧化DNA损伤的情况。结果 与假手术组比较,脓毒症组小鼠前额叶皮层神经元数量显著减少,活性氧含量增加,8-OHdG表达增多(P<0.05);与脓毒症组相比较,参附黄组神经元数量增多,活性氧含量降低,8-OHdG表达减少(P<0.05)。结论 参附黄配方对CLP小鼠前额叶皮质神经元损伤具有一定保护作用,其机制可能降低氧化应激和氧化DNA损伤有关。

Objective To investigate the effect and mechanism of Shenfuhuang Formula(参附黄配方,SF)on neuron injury in sepsis.Methods C57BL/6J mice were randomly divided into sham operation group,sepsis group and SF group,with 26 mice in each group.After anesthesia in the sham operation group,the cecum was left free,without ligation and needling.Cecal ligation and perforation(CLP)was used to induce sepsis model in the sepsis group and SF group.Four hours after modeling,the SF group was given 2.5 g/(kg·d)of SF by gavage,every 12 hours,twice a day for three days,while the sepsis group and the sham operation group were given 0.5 ml of normal sa⁃line by gavage in the same administration way.Seventy-two hours after modeling,Nissl staining was used to observe the changes in the number of neurons in the perfrontal cortex of the brain,and dihydroethidium staining was used to detect active oxygen content in the cerebral cortex tissue;8-Hydroxydeoxyguanosine(8-OHdG)immunofluorescence staining was used to evaluate the oxidative DNA damage in cerebral cortex.Results Compared to those of the sham operation group,the number of neurons in the prefrontal cortex of the sepsis group were reduced,while the content of reactive oxygen species increased,and the expression of 8-OHdG increased(P<0.05).Compared to those of the sepsis group,the number of neurons in the SF group increased,while the content of reactive oxygen species were in⁃hibited,and the expression of 8-OHdG decreased(P<0.05).Conclusion SF has a protective effect on the damage of prefrontal cortex neurons in CLP mice,and its mechanism may be related to the reduction of oxidative stress and oxidative DNA damage.