通络益脑方对急性局灶性脑缺血模型大鼠炎症因子及脑细胞自噬相关蛋白的影响

Effects of Tongluo Yinao Decoction on Brain Histopathology,Inflammatory Factors and Autophagy-related Proteins in Brain of Rats with AFCI

目的观察通络益脑方对急性局灶性脑缺血模型大鼠脑组织病理学、炎症因子及脑细胞自噬相关蛋白的影响。方法将50只雄性SD大鼠随机分为假手术组、模型组、阳性组和试验组(低剂量和高剂量),每组10只。模型组,高、低剂量组和尼莫地平组采用开颅结扎法制备急性局灶性脑缺血大鼠模型,假手术组和模型组给予0.9%氯化钠注射液灌胃,尼莫地平组给予尼莫地平灌胃,高、低剂量组给予通络益脑方灌胃,3周后评估各组大鼠的神经功能评分、神经行为学评分,比较脑梗死体积和神经细胞凋亡。ELISA法检测血清肿瘤坏死因子-α(TNF-α)和白介素-1β(IL-1β)、白介素-2(IL-2)、白介素-6(IL-6)炎症因子水平以及ICAM-1水平,RT-PCR检测PI3K、Akt、Beclin-1以及Bcl-2蛋白相对表达水平。结果与假手术组比较,模型组大鼠的神经功能评分、神经行为学评分、脑梗死体积、神经细胞凋亡、ICAM-1水平、TNF-α、IL-1β、IL-2以及IL-6水平、PI3K、Akt、Beclin-1以及Bcl-2水平明显高于其余3组(P<0.05)。给药后,尼莫地平组和高、低剂量组大鼠的神经功能评分和神经行为学评分、脑梗死体积、神经细胞凋亡、ICAM-1水平、炎症因子水平以及自噬相关蛋白水平明显低于模型组(P<0.05),低剂量组与高剂量组以及尼莫地平组比较均有显著性差异(P<0.05),高剂量组与尼莫地平组比较无显著性差异(P>0.05)。结论通络益脑方对急性局灶性脑缺血大鼠脑组织有较好的保护作用,可减少其脑梗死面积,保护脑组织免受炎症因子的损伤,增加自噬相关蛋白水平,减少神经细胞的凋亡,激活脑细胞自我保护。

Objective:To investigate the effects of Tongluo Yinao Decoction on brain histopathology,inflammatory factors and autophagy-related proteins in brain of rats with acute focal cerebral ischemia(AFCI).Methods:50 male SD rats were randomly divided into the sham group,the model group,nimodipine group and low dose and high dose groups,10 in each.In the model group,the experimental groups and the nimodipine group,AFCI rat model was prepared by craniotomy ligation.The sham group and the model group were given normal saline;the positive group was given nimodipine,and the low dose and high dose groups were given Tongluo Yinao Decoction for gavage.After 3 weeks,the neurological scores and neurobehavioral scores of each group were evaluated,and the volume of cerebral infarction and neuronal apoptosis were compared.TNF-α,inflammatory factor levels,such as IL-1β,IL-2,IL-6,and ICAM-1 levels were detected by Elisa assay,and RT-PCR was used to detect the relative expression levels of PI3 K,Akt,Beclin-1 and Bcl-2 proteins.Results:Compared with the sham group,the neurological function score,neurobehavioral score,cerebral infarction volume,neuronal apoptosis,level of ICAM-1,TNF-α,IL-1β,IL-2,IL-6,PI3 K,Akt,Beclin-1 and Bcl-2 in the model group were significantly higher than the other three groups(P<0.05).After administration,the neurological score and neurobehavioral score,cerebral infarction volume,neuronal apoptosis,ICAM-1 level,inflammatory factor level,and autophagy-related protein levels in the low dose and high dose groups were significantly lower than those in the model group(P<0.05),and there was significant difference among the low-dose group,the high-dose group and the nimodipine group(P<0.05).There was no significant difference between the high dose group and the nimodipine group(P>0.05).Conclusion:Tongluo Yinao Decoction has a good protective effect on brain tissue of rats with acute focal cerebral ischemia,which can reduce the area of cerebral infarction,protect brain tissue from inflammatory factors and increase the level of autophagy-related protein.It reduces the apoptosis of nerve cells and activates the self-protection of brain cells.