培元化瘀方对家兔慢性硬膜下血肿模型的干预作用及其作用机制研究

Intervention Effect and Action Mechanism of Peiyuan-Huayu Recipe in Treating Chronic Subdural Hematoma in Rabbits

目的探讨培元化瘀方对家兔慢性硬膜下血肿(CSDH)模型的干预作用及其作用机制。方法本实验于2018年3—9月完成。采用随机数字表法将造模成功的CSDH家兔21只分为模型组、低剂量组、高剂量组,每组7只。模型组家兔给予0.9%氯化钠溶液灌胃,15ml/次,2次/d;低剂量组家兔给予培元化瘀方3.1g/kg+0.9%氯化钠溶液稀释至15 ml后灌胃,2次/d;高剂量组家兔给予培元化瘀方6.2g/kg+0.9%氯化钠溶液稀释至15 ml后灌胃,2次/d。三组家兔均连续灌胃7d。采用Purty评分评估三组家兔灌胃前及灌胃后第1、3、5、7天神经功能缺损程度;比较三组家兔最后1次灌胃次日残余血肿量;采用HE染色法观察三组家兔血肿外膜及血肿同侧脑组织形态学变化,并采用免疫组化染色法及IPP法检测三组家兔血肿外膜血管内皮生长因子(VEGF)相对表达量。结果(1)时间与方法在Purty评分上存在交互作用(P<0.05);时间在Purty评分上主效应显著(P<0.05);方法在Purty评分上主效应显著(P<0.05)。灌胃后第7天低剂量组、高剂量组家兔Purty评分低于灌胃前(P<0.05);灌胃后第7天高剂量组家兔Purty评分低于模型组、低剂量组,低剂量组家兔Purty评分低于模型组(P<0.05)。(2)高剂量组有1只家兔血肿完全吸收而未成功吸取出硬膜下残余液体。高剂量组家兔残余血肿量少于低剂量组、模型组,低剂量组家兔残余血肿量少于模型组(P<0.05)。(3)HE染色结果显示,与模型组相比,低剂量组、高剂量组家兔血肿外膜纤维组织趋于成熟、炎性反应较轻;模型组家兔血肿同侧脑组织内存活神经元数量少于低剂量组、高剂量组,而低剂量组与高剂量组家兔血肿同侧脑组织内存活神经元数量接近;模型组家兔血肿外膜VEGF相对表达量高于低剂量组、高剂量组(P<0.05),而低剂量组与高剂量组家兔血肿外膜VEGF相对表达量比较,差异无统计学意义(P>0.05)。结论培元化瘀方可有效减轻CSDH家兔神经功能缺损程度并促进血肿吸收,其作用机制可能与抑制血肿及血肿外膜炎性反应、VEGF的表达、受压脑组织神经元坏死及凋亡等有关。

Objective To investigate the intervention effect and action mechanism of Peiyuan-Huayu recipe in treating chronic subdural hematoma(CSDH)in rabbits.Methods This experiment was conducted from March to September 2018.A total of 21 CSDH rabbits with successful modeling were divided into model group,low-dose group and high-dose group according to random number table method,with 7 rabbits in each group.Rabbits in model group received intragastric administration of 0.9%sodium chloride solution(15 ml per time,2 times per day),rabbits in low-dose group received intragastric administration of low-dose Peiyuan-Huayu recipe(3.1 g/kg,2 times per day,diluted to 15 ml by 0.9%sodium chloride solution),while rabbits in high-dose group received intragastric administration of high-dose Peiyuan-Huayu recipe(6.2 g/kg,2 times per day,diluted to 15 ml by 0.9%sodium chloride solution);all of the three groups continuously received intragastric administration for 7 days.Purty score was used to evaluate the degree of neurological deficit before intragastric administration,1 day,3 days,5 days and 7 days after intragastric administration;residual hematoma volume next day after the last intragastric administration was compared in the three groups;HE staining method was used to observe the morphological change of hematoma adventitia and ipsilateral brain tissue of hematoma,meanwhile immunohistochemistry assay and IPP method were used to detect the relative expression quantity of VEGF in hematoma adventitia.Results(1)There was statistically significant interaction between time and method in Purty score(P<0.05);main effects of time and method were statistically significant(P<0.05).After 7 days of intragastric administration,Purty score in low-dose group and high-dose group was statistically significantly lower than that before intragastric administration,respectively(P<0.05),Purty score in high-dose group was statistically significantly lower than that in model group and low-dose group,respectively,meanwhile Purty score in low-dose group was statistically significantly lower than that in model group(P<0.05).(2)In high-dose group,1 rabbit did not successfully extract subdural residual fluid due to complete absorption of hematoma.Residual hematoma volume in highdose group was statistically significantly less than that in model group and low-dose group,respectively,meanwhile residual hematoma volume in low-dose group was statistically significantly less than that in model group(P<0.05).(3)HE staining results showed that,fibrous tissue tended to maturity and inflammatory reaction was milder in hematoma adventitia in low-dose group and high-dose group compared to that in model group,respectively;number of surviving neurons in ipsilateral brain tissue of hematoma in model group was less than that in low-dose group and high-dose group,while number of surviving neurons in ipsilateral brain tissue of hematoma was similar between low-dose group and high-dose group;relative expression quantity of VEGF in hematoma adventitia in model group was statistically significantly higher than that in low-dose group and high-dose group(P<0.05),while no statistically significant difference of relative expression quantity of VEGF in hematoma adventitia was found between low-dose group and high-dose group(P>0.05).Conclusion Peiyuan-Huayu recipe can effectively reduce the degree of neurological deficit and promote the absorption of hematoma in rabbits with CSDH,its action mechanism may correlate with the inhibition of inflammatory reaction and expression of VEGF in hematoma and hematoma adventitia,necrosis and apoptosis of neurons in pressurized brain tissue.