脑血疏口服液不同用药时点对脑出血急性期大鼠血肿大小及CD36表达的影响

Effect of Different Medication Time Points of Naoxueshu Oral Liquid on theSize of Hematoma and CD36 Expression in Rats with Acute Cerebral Hemorrhage

目的:观察脑血疏口服液在不同用药时点对脑出血急性期大鼠血肿大小及CD36表达的影响。方法:采用尾状核注射Ⅶ型胶原酶构建大鼠脑出血模型。将40只SD大鼠随机分为假手术组(A组)、模型组(B组)、预给药组(C组)、造模后6 h给药组(E组)、造模后24 h给药组(G组)。造模前,C组给予脑血疏口服液灌胃,其余各组给予等量生理盐水灌胃,共7 d。造模后,C、E和G组分别于各自时点给予脑血疏口服液灌胃,其余各组给予等量生理盐水,共3 d。运用改良的神经功能缺损评分标准(mNSS)评估大鼠神经功能缺损程度,磁共振成像技术(MRI)检测脑内血肿体积,苏木-伊红染色(HE)观察血肿周围组织形态,透射电镜观察血肿周围组织超微结构,免疫组化(IHC)观察血肿周围组织CD36阳性细胞数,蛋白印迹法(Western Blot)检测CD36蛋白表达。结果:神经功能缺损评分结果显示,各组大鼠在造模后均出现不同程度的神经功能缺损,与B组相比,C组及E组神经功能评分降低(P<0.05)。病理染色结果提示,C、E、G组较B组血肿周围组织坏死面积减少,周围组织疏松减轻,神经元超微结构改善,线粒体数量增多。MRI检测显示,与B组相比,C、E、G组大鼠血肿体积减小(P<0.05),其中,C、E组血肿吸收率显著提高(P<0.05)。Western Blot结果提示,与B组相比,C、E组的CD36表达水平显著升高(P<0.05)。免疫组化结果提示,与B组相比,C、E、G组大鼠的血肿周围组织CD36免疫标记阳性表达染色加深,数量增多,CD36表达水平升高(P<0.05)。结论:脑出血急性期应用脑血疏口服液不会增加血肿扩大风险及再出血风险,在不同时点给药均可改善大鼠神经功能,促进大鼠血肿吸收,减轻神经元结构损害,且在脑出血急性期6 h内给药疗效最佳,其机制可能与上调大鼠脑出血后CD36的表达有关。

Objective:To observe the effect of different medication time points of naoxueshu(NXS)oral liquid on the size of hematoma and CD36 expression in rats with acute cerebral hemorrhage.Methods:The rat model of cerebral hemorrhage was established by injecting collagenaseⅦinto caudate nucleus.Forty SD rats were randomly divided into sham operation group(group A),model group(group B),pre-administration group(group C),6 h post-modeling administration group(group D),and 24 h post-modeling administration group(group E).Before modeling,group C was gavaged with NXS oral liquid and other groups with the same amount of normal saline for 7 days.After modeling,groups C,E and G were garaged with NXS oral liquid at their respective time points and the other groups with the same amount of normal saline at the same time points for 3 d.Modified neurological severity score(mNSS)was used to evaluate the degree of neurological impairment in the rats,and magnetic resonance imaging(MRI)technology was used to measure the volume of the hematoma.Hematoxylin-eosin(HE)staining was used to observe tissue morphology around hematoma,and transmission electron microscopy was applied to observe the ultrastructure of tissues around hematoma.Immunohistochemistry(IHC)was used to observe the number of CD36 positive cells in the surrounding tissue of the hematoma,and Western blot was used to detect CD36 expression.Results:The modified neurological severity score showed that all rats had neurological impairment after modeling,and the neurological function scores of groups C and E were lower than those of group B(P<0.05).Pathological staining indicated that compared with group B,the areas of tissue necrosis around hematoma in group C,E and G were reduced,surrounding tissue looseness was reduced,the ultrastructure of neurons was improved,and the number of mitochondria was increased.MRI showed that compared with group B,the volume of hematoma in groups C,E and G decreased(P<0.05),and the hematoma absorption rate in groups C and E was significantly increased(P<0.05).Western blot results indicated that,compared with group B,the expression level of CD36 in groups C and E was significantly increased(P<0.05).The Immunohistochemistry results indicated that,compared with group B,the CD36 positive expression staining was deepened,the number was increased and the expression level of CD36 was increased in the perihematoma tissues of the rats in groups C,E and G(P<0.05).Conclusion:Application of NXS oral liquid does not increase the risks of hematoma enlargement and rebleeding in the acute stage of intracerebral hemorrhage.Drug administration at different time points can improve the nerve function of the rats,promote the absorption of the hematoma and reduce the damage of the neuron structure,and the administration of drugs within 6 hours after acute intracerebral hemorrhage produces the best effect.The mechanism may be related to the up-regulation of CD36 expression after intracerebral hemorrhage.