活血、利水中药对脑出血大鼠脑组织肿瘤坏死因子-α、核转录因子-κB及水通道蛋白-4表达的影响

目的探讨活血、利水中药对脑出血急性期大鼠脑组织肿瘤坏死因子-α(tumor necrosis factor-al-pha,TNF-α)、核转录因子-κB(nuclear factor-kappaB,NF-κB)及水通道蛋白-4(aquaporin-4,AQP-4)表达的影响及其治疗脑水肿的可能机制。方法自体股动脉血注入脑尾状核建立大鼠脑出血模型,SD雄性大鼠随机分为假手术组(42只)、模型组(42只)、活血组(42只)和利水组(42只),其中活血组予活血中药复方(每毫升含生药量分别为:大黄0.2g、水蛭0.02g、三七0.3g)10mL/kg灌胃,1次/日;利水组予利水中药复方(每毫升含生药量分别为:茯苓0.2g、泽泻0.2g、石菖蒲0.2g)10mL/kg灌胃,1次/日。模型组和假手术组予生理盐水4.0mL/kg灌胃,1次/日。1、3、5d取材,免疫组织化学染色法和实时荧光定量PCR法检测大鼠脑组织TNF-α、NF-κB p65和AQP-4mRNA和蛋白的表达。结果与假手术组比较,模型组大鼠各时间点脑组织TNF-α、NF-κBp65和AQP-4蛋白及mRNA表达明显增加(P<0.01,P<0.05);与模型组比较,活血组和利水组各时间点AQP-4和TNF-α表达显著降低(P<0.05);活血组NF-κB p65表达亦明显低于模型组(P<0.05),利水组NF-κB p65表达差异无统计学意义(P>0.05);与模型组比较,活血组和利水组各时间点脑组织含水量均有不同程度的降低,活血组与利水组间比较,差异亦有统计学意义(P<0.05)。结论活血、利水中药均可抑制炎性因子TNF-α的释放,下调AQP-4的表达,减轻大鼠血肿周围脑水肿,但作用强度和作用环节有差异。

Effects of Herbs Capable of Activating Blood Circulation or Inducing Diuresis on the Expressions of Tumor Necrosis Factor-alpha,Nuclear Factor-kappaB,and Aquaporin-4 in Rats with Intracerebral Hemorrhage

Objective To investigate the effects of herbs capable of activating blood circulation or inducing diuresis on the expressions of tumor necrosis factor-alpha(TNF-α),nuclear factor-kappaB(NF-κB),and aquaporin-4(AQP-4) in rats with intracerebral hemorrhage(ICH),and to study their possible mechanisms.Methods The ICH rat model was established by injecting autologous arterial blood into the right caudate nucleus.The 168 male rats were randomly divided into four groups,the sham-operative group,the model group,the blood activating group,and the diuresis inducing group,42 in each group.Chinese compound decoction(consisting of 0.2 g rhubarb,0.02 g leech,and 0.3 g notoginseng in each milliliter decoction) was given to rats in the blood activating group by gastrogavage at the dose of 10 mL/kg,once daily.Chinese compound decoction(consisting of 0.2 g poria,0.2 g water plantain tuber,and 0.2 g acori graninei in each milliliter decoction) was given to rats in the diuresis inducing group by gastrogavage at the dose of 10 mL/kg,once daily.4.0 mL/kg normal saline was given to rats in the model group and the sham-operative group by gastrogavage,once daily.A series of brain samples were obtained on the 1st,3rd,and 5th day,respectively.The mRNA and protein expressions of TNF-α,NF-κB p65,and AQP-4 were determined by immunohistochemical staining and Real-time fluorescent quantitative PCR respectively.Results After ICH,TNF-α,NF-κB,and AQP-4 protein positive cells in the brain tissue and their protein and mRNA expressions significantly increased in rats of the model group at each time point when compared with the sham-operative group(P<0.01,P<0.05).The gene and protein expressions of TNF-α and AQP-4 significantly decreased in the blood activating group and the diuresis inducing group at each time point when compared with the model group(P<0.05).The expression of NF-κB p65 in the blood activating group obviously decreased when compared with the model group(P<0.05),but there was no statistical difference in the NF-κB expression when compared with the diuresis inducing group(P>0.05).Compared with the model group,the water content of the brain tissue decreased to some degree in the blood activating group and the diuresis inducing group at each time point.There was statistical difference between the blood activating group and the diuresis inducing group(P<0.05).Conclusions Chinese herbs capable of activating blood circulation or inducing diuresis could inhibit the release of TNF-alpha,down-regulate the expression of AQP-4,and alleviate the brain edema around hematoma.But the action strength and the effect links were different.