桃红四物汤对脑缺血再灌注大鼠海马及皮质的影响

Taohong Siwu Tang Has Effect on Hippocampus and Cortex in Rats with Cerebral Ischemia-Reperfusion

目的:观察桃红四物汤(TSD)对脑缺血再灌注(CIR)损伤大鼠海马及皮质的影响,并对其机制进行初步探讨。方法:将60只雄性Wistar大鼠分为假手术组(Sham组)、模型组(Vehicle组)、TSD低、中、高剂量组及阳性药组(EDA组)。TSD低、中、高剂量组分别灌胃5 g/kg、10 g/kg、20 g/kg的TSD,EDA组灌胃4 mg/kg的依达拉奉(EDA)混悬液,Sham组及Vehicle组灌胃等量生理盐水,连续给药5天。除Sham组外,其余各组制备中动脉阻塞(MCAO)模型,脑缺血后24 h处死大鼠。模型制备成功后,对各组大鼠进行神经功能缺失评分,对大鼠大脑组织进行2,3,5-氯化三苯基四氮唑(TTC)染色,Western blot法测定大鼠受损侧海马及皮质中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达。结果:与Sham组比较,Vehicle组大鼠神经功能缺失评分、半球肿胀率及脑梗死体积明显升高(P<0.05),海马及皮质区IL-1β、IL-6、TNF-α、Bax表达上升(P<0.05),Bcl-2蛋白表达下降(P<0.05)。与Vehicle组比较,EDA组及TSD中、高剂量组大鼠神经功能缺失评分、半球肿胀率及脑梗死体积降低(P<0.05),EDA组及TSD高剂量组海马及皮质区IL-1β、IL-6、TNF-α、Bax表达下降(P<0.05),Bcl-2蛋白表达升高(P<0.05)。与EDA组比较,TSD高剂量组大鼠神经功能缺失评分、半球肿胀率及脑梗死体积降低(P<0.05),海马及皮质区IL-1β、Bax表达降低,Bcl-2表达升高(P<0.05)。结论:TSD能够显著改善MCAO模型大鼠神经功能损伤,实现脑神经保护,其作用机制涉及炎症及凋亡蛋白信号通路。

Objective:To observe the effect of Taohong Siwu tang(TSD)on hippocampus and cortex in rats with cerebral ischemia-reperfusion(CIR)injury,and to carry out a preliminary study on its mechanism.Methods:A total of 60 male Wistar rats were divided into the sham operation group(the Sham group),the model group(the Vehicle group),the TSD groups of low-dose,medium-dose and high-dose,and the positive medicine group(the EDA group).The TSD groups of low-dose,medium-dose and high-dose were given gastric perfusion of TSD of 5 g/kg,10 g/kg and 20 g/kg.The EDA group was given gastric perfusion of edaravone(EDA)suspension at a dose of 4 mg/kg.The Sham group and the Vehicle group were given saline of the same volume by gavage.All groups were continuously administrated for five days.Except for the Sham group,in the other groups,rats were sacrificed after cerebral ischemia for 24 hours to establish middle cerebral artery occlusion(MCAO)model.After successful establishment,the score of neurological deficits was evaluated;the cerebral tissues of rats were stained with 2,3,5-triphenyltetrazolium chloride(TTC);the expression of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),B-cell lymphoma 2(Bcl-2),and Bcl-2 associated X(Bax)in the hippocampus and cortex of the damaged side was detected by Western blot method.Results:When compared with those in the Sham group,in the Vehicle group,the score of neurological deficits,the rate of cerebral hemispheric swelling,and the volume of cerebral infarction were significantly increased(P<0.05);the expression of IL-1β,IL-6,TNF-αand Bax in hippocampus and cortical area was increased(P<0.05);the protein expression of Bcl-2 was decreased(P<0.05).When compared with those in the Vehicle group,in the EDA group and the TSD groups of medium-dose and high-dose,the score of neurological deficits,the rate of cerebral hemispheric swelling,and the volume of cerebral infarction were decreased(P<0.05);in the EDA group and the high-dose TSD group,the expression of IL-1β,IL-6,TNF-αand Bax in hippocampus and cortical area was decreased(P<0.05);the protein expression of Bcl-2 was increased(P<0.05).When compared with those in the EDA group,in the high-dose TSD group,the score of neurological deficits,the rate of cerebral hemispheric swelling,and the volume of cerebral infarction were decreased(P<0.05);the expression of IL-1βand Bax in hippocampus and cortical area was decreased(P<0.05);the expression of Bcl-2 was increased(P<0.05).Conclusion:TSD can significantly improve the impairment of nerve function of MCAO rat models,and protect the cerebral nerve,whose mechanism is related to the signaling pathway of inflammatory and apoptosis proteins.