出血性中风1号方对脑出血大鼠脑组织的影响

Effect of First Prescription for Hemorrhagic Stroke on the brain tissue of rats with intracerebral hemorrhage

目的 研究出血性中风1号方对脑出血(ICH)大鼠脑组织的影响。方法 选取84只雄性SD大鼠,体重(220±20)g,随机选取24只,采用随机数字表法将其分为空白组和假手术组(注射生理盐水),每组12只。剩余60只采用胶原酶Ⅶ注射诱导ICH模型,造模成功后采用随机数字表法将其分为模型组、脑血疏组(2.08 ml/kg)及出血性中风1号方低、中、高剂量组(3.2、6.4、12.8 g/kg)。空白组、假手术组、模型组灌胃等量生理盐水,其余组灌胃对应剂量药物,1次/d,连续7 d。给药结束后,采用苏木精-伊红染色观察七组病理学变化;干湿重法测定七组脑含水量变化;采用酶联免疫吸附试验检测七组肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平,Western blot检测核因子κB(NB-κB)p65及磷酸化NB-κB p65(p-NF-κB p65)水平。结果 空白组、假手术组脑组织结构完整,模型组有大量红细胞及炎症细胞浸润,排列紊乱,水肿严重,出血性中风1号方低、中、高剂量组均有少量红细胞。空白组脑含水量、TNF-α、IL-6、p-NF-κB p65水平与假手术组比较,差异均无统计学意义(P> 0.05)。模型组脑含水量、TNF-α、IL-6、p-NF-κB p65高于空白组(P <0.05)。出血性中风1号方中、高剂量组脑含水量、TNF-α、IL-6、p-NF-κB p65低于模型组和出血性中风1号方低剂量组(P <0.05)。出血性中风1号方低剂量组脑含水量、TNF-α、IL-6、p-NF-κB p65水平与模型组比较,差异无统计学意义(P> 0.05)。出血性中风1号方中、高剂量组脑含水量、TNF-α、IL-6、p-NF-κB p65比较,差异无统计学意义(P> 0.05)。各组NF-κB p65水平比较,差异无统计学意义(P> 0.05)。结论 中、高剂量的出血性中风1号方对ICH大鼠有脑保护作用,可能与抑制p-NF-κB p65、TNF-α、IL-6表达有关。

Objective To study the effect of First Prescription for Hemorrhagic Stroke on the brain tissue of rats with intracerebral hemorrhage(ICH). Methods A total of 84 male SD rats with the body weight of(220±20) g were randomly selected and divided into blank group and sham operation group(injected with normal saline) by random number table method, with 12 rats in each group. The remaining 60 rats were treated with ICH model induced by collagase Ⅶ injection, and were divided into model group, Naoxueshu group(2.08 ml/kg), and First Prescription for Hemorrhagic Stroke low-dose, medium-dose, and high-dose groups(3.2, 6.4, 12.8 g/kg) by random number table method. Blank group,sham operation group, and model group were intragastric with equal amount of normal saline, and the other groups were intragastric with corresponding dose of drugs, once a day, for consecutive seven days. After administration, hematoxylin-eosin staining was used to observe the pathological changes of seven groups. The changes of cerebral water content in seven groups were determined by wet and dry weight method. The levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6) in the seven groups were detected by enzyme linked immunosorbent assay, and the levels of nuclear factor-κB(NB-κB) p65 and phosphorylated NF-κB(p-NF-κB) p65 were detected by Western blot.Results The blank group and the sham operation group had complete brain tissue structure, the model group had a large number of red blood cells and inflammatory cells infiltration, disorder, severe edema, First Prescription for Hemorrhagic Stroke low-dose, medium-dose, and high-dose groups all had a small number of red blood cells. There were no significant differences in cerebral water content, TNF-α,IL-6, and p-NF-κB p65 levels between blank group and sham group(P > 0.05). The cerebral water content, TNF-α,IL-6, and p-NF-κB p65 in model group were higher than those in blank group(P < 0.05). The cerebral water content,TNF-α, IL-6, and p-NF-κB p65 in First Prescription for Hemorrhagic Stroke medium-dose and high-dose groups were lower than those in the model group and First Prescription for Hemorrhagic Stroke low-dose group(P < 0.05).There were no significant differences in cerebral water content, TNF-α, IL-6, and p-NF-κB p65 levels between First Prescription for Hemorrhagic Stroke low-dose group and model group(P > 0.05). There was no significant difference in cerebral water content, TNF-α, IL-6, and p-NF-κB p65 between Hemorrhagic Stroke medium-dose and high-dose groups(P > 0.05). There was no significant difference in NF-κB p65 level among all groups(P > 0.05). Conclusion Medium and high dose of First Prescription for Hemorrhagic Stroke has brain protective effect on ICH rats, which may be related to the inhibition of p-NF-κB P65, TNF-α, and IL-6 expression.