基于调节炎症因子研究醒脑灌肠液对ICH大鼠的脑保护作用

Study on the Cerebral Protection of Xingnao Enema in Rats with Intracerebral Hemorrhage Based on the Regulation of Inflammatory Factors

目的:探讨醒脑灌肠液对脑出血大鼠不同时间点炎症因子的调节作用,研究其脑保护作用机制。方法:建立脑出血(ICH)模型,将Wistar雄性大鼠随机分成6组:空白组、假手术组、模型对照组、安宫牛黄丸0.27 g/kg组、醒脑灌肠液1.80 g/kg和3.60 g/kg组。各组大鼠分别灌肠给予对应的药物,在给药12 h、24 h、3 d、7 d后,采集样本。ELISA法检测出血侧脑组织TNF-ɑ、IL-6、IFN-γ、IL-4和IL-10的含量,Western blot法检测脑组织TSG-6蛋白的表达,TUNEL法检测神经细胞凋亡的情况。结果:与假手术组相比,在4个时间点,模型组脑组织中TNF-α、IL-6和IFN-γ的含量均显著升高,IL-4、IL-10的含量在出血后初期先升高后降低,脑组织TSG-6的蛋白表达显著下调,细胞凋亡指数在第3 d时达高峰。与模型组相比,安宫牛黄丸和醒脑灌肠液各剂量组均能够显著降低TNF-α、IL-6、IFN-γ的含量以及细胞凋亡指数,显著升高IL-4、IL-10的含量,上调TSG-6蛋白的表达。结论:脑出血后,醒脑灌肠液通过调节细胞因子的释放,抑制炎症反应,抑制神经细胞的凋亡,发挥脑保护作用。

Objective:To explore the regulating mechanism of Xingnao enema on the inflammatory factors at different time points in rats with intracerebral hemorrhage(ICH)and its cerebral protection mechanism Methods:Intracerebral hemorrhage(ICH)model was established.Wistar rats were randomly divided into the control group,the sham-operated group,the model group,Angong Niuhuang pill group(0.27 g/kg)and Xingnao enema groups(3.60 g/kg,1.80 g/kg).All rats were administered with corresponding drugs,and samples were collected at 12 h,24 h,3 d and 7 d after enema.The contents of TNF-α,IL-6,IFN-γ,IL-4,and IL-10 in the hemorrhagic lateral brain tissues were detected by ELISA,the expression of TSG-6 was detected by western blot,the neuron apoptosis in the brain tissue was detected by TUNEL assay.Results:Compared with the sham-operated group,the contents of TNF-αand IL-6 in the model group were significantly increased at 4 time points,the contents of IL-4,IL-10 were increased at 12 h,then decreased after 12 h,the expression of TSG-6 was significantly decreased,the apoptosis index of nerve cells reached the increasing peak on the 3 rd day.Compared with the model group,after the intervention of Angong Niuhuang pill and Xingnao enema,the contents of TNF-α,IL-6,IFN-γand the apoptosis index of nerve cells in the hemorrhagic lateral brain tissues were significantly decreased,while the contents of IL-4,IL-10 and the expression of TSG-6 were significantly increased.Conclusion:After ICH,Xingnao enema can protect the brain by regulating the release of cytokines,inhibiting inflammatory reaction and the apoptosis of the nerve cells.