安宫牛黄丸对脓毒症脑血管vWF表达的调控机制研究

Study on the Regulatory Mechanism of Angong Niuhuang Pills on the Expression of vWF in Septic Cerebrovascular

目的 基于血管性血友病因子(vWF)相关调控关系,观察安宫牛黄丸在脓毒症脑血管损伤中的作用。方法 选用C57BL/6J小鼠60只,随机分为对照组、模型组、安宫牛黄丸低剂量组(1.0 g/kg)、安宫牛黄丸中剂量组(2.0 g/kg)、安宫牛黄丸高剂量组(3.0 g/kg)、右美托咪定组。腹腔注射20 mg/kg脂多糖(LPS)复制脓毒症模型。苏木素-伊红(HE)染色观察小鼠脑组织的病理学改变;干湿重比重法检测各组小鼠脑组织含水量;ELISA法检测各组小鼠血清中v WF表达水平;免疫组织化学法观察小鼠脑血管中v WF阳性表达水平;Western blotting检测小鼠脑血管中vWF蛋白表达水平;实时荧光定量PCR(qRT-PCR)检测小鼠脑血管中vWF mRNA表达水平。结果 与模型组相比,各用药组小鼠海马神经细胞排列较整齐,神经细胞核固缩深染情况明显改善,组织疏松、水肿减轻;安宫牛黄丸可显著降低小鼠脑含水量(P <0.01);安宫牛黄丸各治疗组小鼠血清中vWF含量显著降低(P <0.01),脑血管中vWF阳性表达明显降低;脑血管中v WF蛋白表达显著降低(P <0.01),脑血管中vWF mRNA表达显著降低(P <0.01)。结论 安宫牛黄丸可以降低脓毒症小鼠中血清中vWF的含量及脑血管中vWF的表达,减轻脑水肿,对脓毒症小鼠脑组织起到保护的作用。

Objectives:To investigate the effect of Angong Niuhuang Pills on septic cerebrovascular injury based on the related regulatory relationship of von Willebrand Factor(v WF). Methods:60 C57BL/6J mice were selected and randomly divided into the control group,lipopolysaccharide(LPS)model group,Angong Niuhuang Pills low dose(AG-L)group(1.0 g/kg),Angong Niuhuang Pills medium dose(AG-M)group(2.0 g/kg),Angong Niuhuang Pills high dose(AG-H)group(3.0 g/kg),and Dexmedetomidine(Dex)group. The sepsis model was established by intraperitoneal injection of 20 mg/kg LPS. Hematoxylin-eosin(HE)staining was applied to observe the pathological changes of mouse brain tissue;determination of water content in brain tissue of mice in each group was performed by dry-wet specific gravity method;ELISA method was used to detect the expression level of vWF in the serum of mice in each group;immunohistochemical method was used to observe the positive expression of vWF in the cerebral blood vessels of mice;Western blotting was used to detect the expression level of v WF protein in mouse cerebral blood vessels;real-time quantitative PCR(qRT-PCR)was used to detect the expression level of vWF m RNA in mouse cerebral blood vessels. Results:Compared with the LPS group,the hippocampal nerve cells of the mice in each drug group were arranged neatly,the pyknosis and hyperchromasia of the nerve cells were significantly improved,and the tissue was loose and the edema was reduced;Angong Niuhuang Pills could significantly reduce the water content of brain in mice(P < 0.01);the content of vWF in serum of mice in Angong Niuhuang Pills treatment group was significantly decreased(P < 0.01);the positive expression of vWF in cerebral blood vessels was significantly decreased,the expression of v WF protein in cerebral blood vessels was significantly decreased(P < 0.01),the expression of vWF m RNA in cerebral blood vessels was significantly decreased(P < 0.01). Conclusion:Angong Niuhuang Pills can reduce the content of vWF in serum and the expression of vWF in cerebral blood vessels in septic mice,reduce cerebral edema,protect the brain tissue of mice with sepsis.