摘要
目的探讨凝血因子Ⅶ（FⅦ）与小鼠进展性脑挫裂伤出血的关系，为重组人FⅦa的临床应用提供实验依据。方法（1）12只BALB／c雄性小鼠分别给予1，3，5，10mg／kg的脂质体包裹的FⅦsiRNA注射动物尾静脉，每剂量组3只，另3只注射等渗盐水，注射后2d予眼球取血，PCR检测肝脏FVII的表达，ELISA法检测血浆FⅦ浓度，底物显色法检测血浆FⅦ活性；选择最佳剂量的脂质体包裹的FⅦsiRNA来抑制小鼠FⅦ的表达。（2）30只BALB／c雄性小鼠按随机数字表法分为FⅦ抑制组和对照组，FⅦ抑制组给予最佳剂量的脂质体包裹的FⅦsiRNA；对照组则给予同等剂量的脂质体包裹的阴性对照siRNA，每组15只，均建立脑挫裂伤出血模型，于伤后3，24，72h检测两组脑挫裂伤出血量；伤后24，48h观察两组血肿体积。结果（1）1，3，5，10mg／kg脂质体包裹的FⅦsiRNA抑制后，肝脏FⅦ的表达均明显下降，血浆FⅦ浓度和活性也明显下降。FⅦsiRNA的最佳剂量为3mg／kg。（2）FⅦ抑制组伤后3，24，72h相对脑出血量分别为1．46±0．10，1．82±0．23，2．28±0．15，均显著大于对照组（1．00±0．25，1．20±0．31，1．20±0．22）（P〈0．05）。伤后24，48hFⅦ抑制组血肿体积分别为（6．7±1．5）mm^3、（9．8±1．0）mm^3，均较对照组[（5．2±1．2）mm^3、（5．5±1．5）mm^3]明显增加（P〈0．01）。结论体内FⅦ浓度与进展性脑挫裂伤出血的发生密切相关，脑挫裂伤出血后给予FⅦ可以显著降低进展性脑挫裂伤出血的发生。
Objective To study the correlation between the coagulation factor Ⅶ （F Ⅶ ） and progressive hemorrhage after brain contusion in mice and provide the experimental evidence for the clinical application of recombinant human FⅦa. Methods Twelve male BALB/c mice were given liposome- encapsulated FV^siRNA via tail vein at doses of 1,3, 5 and 10 mg/kg with 3 mice per dosage. The other 3 mice received equivalent volume of normal saline as controls. Two days after the injection, mice blood sampling was used to detect F Ⅶ mRNA expression in liver using real-time PCR, level of plasma F V~ using ELISA method, and activity of plasma FⅦ using chromogenic substrate assay. The optimal dose at which F Ⅶ expression was inhibited was determined. Thirty BALB/c male mice were assigned to two groups （n = 15 per group） according to the random number table： FⅦ-suppressing group, mice were injected with FⅦsiRNA at the optimal dose and control group, mice were injected with same volume of negative control vector. The model of brain contusion was established in both groups. Volume of hemorrhage following brain contusion was measured at 3, 24 and 72 h postinjury, and hematoma volume at 24 and 48 h postinjury. Results Liposome-encapsulated siRNA delivery down-regulated FⅦ expression in the mouse liver. Level and activity of plasma FⅦ were also reduced significantly. The optimal siRNA dose was 3 mg/kg. At 3, 24 and 72 h postinjury, relative volume of brain hemorrhage in FⅦ-suppressing group was 1.46 20.10, 1.82 20.23 and 2.28 20.15 respectively,significantly higher than that in control group（ 1.00 20.25, 1.20 20.31 and 1.20 20.22 respectively） （P 〈0.05）. At 24 and 48 h postinjury, volume of hematoma in FⅦ-suppressing group was （6.7 ± 1.5 ） mm^3 and （9.8 ±1.0） mm^3, significantly higher than that in control group [（5.2 ±1.2）mm^3 and （5.5 ±1.5）mm^3] （P 〈0.01）. Conclusions Level of FⅦ in vivo relates closely to the progressive hemorrhage of brain contusion in mice. Administra
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2015年第11期1009-1013,共5页
Chinese Journal of Trauma
基金
国家自然科学基金资助项目（81471241,81271375,81171133）
关键词
因子Ⅶ
脑损伤
脑出血
Factor Ⅶ
Brain injuries
Cerebral hemorrhage
