参芍口服液对实验性脑出血大鼠脑内CD14/TLR4介导的继发性炎症损伤的干预效果研究

Effect of Shenshao Oral Liquid on CD14/TLR4 Mediated Secondary Inflammatory Injury in the Brain of Rats with Experimental Cerebral Hemorrhage

目的:研究参芍口服液对实验性脑出血大鼠脑内CD14/TLR4介导的继发性炎症损伤的干预效果。方法:实验大鼠随机分为假手术组(Sham组),脑出血模型组(ICH组),参芍口服液干预组(SS组)。每组又分为6h,12h,24h,72h和7d 5个小组。ICH组和SS组用自体股动脉血法复制脑出血模型。SS组苏醒后给予参芍口服液(8mL/d)灌胃。观察脑组织病理形态学变化及未受损神经元计数;干湿重法测定脑含水量变化;免疫组化和免疫印迹法检测CD14、TLR4和IL-8阳性细胞数和蛋白表达水平。结果:ICH组脑出血后6h开始出现水肿,12~72h脑组织含水量逐渐增加,72h达到高峰,之后脑水肿逐渐减弱,7d基本恢复正常;SS组较模型组脑含水量显著降低(P<0.05)。Sham组大鼠CD14在脑组织细胞微量表达;ICH组术后6h血肿周围CD14蛋白开始表达,可见少量阳性细胞随时间延续,表达量逐渐增加,72h蛋白表达达最高峰,以后逐渐降低。TLR4、IL-8蛋白表达水平趋势和CD14相一致。TLR4与CD14表达水平变化与脑含水量呈正相关(P<0.05);参芍口服液干预后,CD14表达水平TLR4、IL-8表达量也显著降低。结论:参芍口服液可能通过减少CD14/TLR4信号通路的诱发因素,降低脑出血后血肿周围脑组织继发性炎症损伤,对脑组织起保护作用。

Objective:To research Shenshao Oral Liquid Inhibits Secondary Inflammatory Injure Induced by CD14/TLR4 after Intracerebral Hemorrhage in Experimental Rats.Methods:Experimental rats were randomly divided into sham operation group,model group and intervention group.Each group further was subdivided into 6 h group,12 h group,24 h group,72 h group,and 7 d group.ICH group and SS group were used to duplicate the model of cerebral hemorrhage.SS group was given Shenshao oral liquid(8ml/d)by gavage after waking up.The number and protein expression of CD14,TLR4 and IL-8 positive cells were detected by immunohistochemistry and Western blotting.Results:The edema was appeared after 6h after the ICH model was established,and the water content of brain was gradually increased at 12h to 72h,and reached to the highest level at 72h,then the brain edema was gradually decreased,and ultimately got right original level at 7d.Compared with the ICH group,the water content of brain in the rats of the intervention group was obviously decreased(P<0.05).The expression of CD14 in brain tissue cells of sham group was slight;CD14 protein began to express around hematoma in ICH group at 6h after operation,and a small number of positive cells increased with time,reached the peak at 72h,and then decreased gradually.The trend of TLR4 and IL-8 protein expression was consistent with that of CD14.The expression level of TLR4 and CD14 was positively correlated with brain water content(P<0.05),and the expression level of TLR4 and IL-8 was significantly decreased after Shenshao oral liquid.Conclusion:Shenshao oral liquid may reduce the inducing factors of CD14/TLR4 Signal pathway,reduce the secondary inflammatory injury of brain tissue around hematoma after cerebral hemorrhage,and play a protective role in brain tissue.