重组人肿瘤坏死因子相关弱凋亡诱导因子对小鼠主动脉内皮舒张功能及凝血相关因子的影响

Impact of Recombinant Human Tumor Necrosis Factor-like Weak Inducer of Apoptosis on the Aorta Endothelial Function and Blood Coagulation

目的采用重组人肿瘤坏死因子相关弱凋亡诱导因子（rhTWEAK）经腹腔注射，诱导血管内皮损伤模型，观察rhTWEAK对小鼠主动脉内皮舒张功能和凝血相关因子的影响。方法3～4周龄C57BL小鼠32只，随机分成4组，每组8只，腹腔注射各药物：@rhTWEAK组：rhTWEAK按5μg/（kg·d）的量溶于生理盐水，共0．3mL，腹腔注射，连续注射7天；②rhTWEAK-4-抗rhTWEAK组：先给予抗rhTⅣEAK腹腔注射，15min后再给予rhTWEAK注射；③IgG组：给予小鼠非特异性IgG腹腔注射；④对照组：给予0．3mL生理盐水注射。分别在干预1周后眼球取血，分离上清；取小鼠胸主动脉段进行血管舒张功能的检测。酶联免疫吸附法检测血浆中内皮型一氧化氮合酶（eNOS）、高敏C反应蛋白（hs·CRP）、一氧化氮（NO）、组织因子（TF）、组织因子途径抑制物（TFPI）含量。结果①与对照组、rhTWEAK＋抗rhTWEAK组、IgG组相比，rhTWEAK组内皮依赖性血管舒张功能、血浆中eNOS、NO、TFPI水平明显降低，TF水平明显升高，均有统计学意义（均P〈0．05）；血浆中hs—CRP含量未见明显改变（P〉0．05）；②rhTWEAK组、IgG组与对照组相比，血管内皮舒张功能、血浆中NO、TF、TFPI、eNOS、hs—CRP含量变化均无统计学差异（均P〉0．05）。结论rhTWEAK可诱导内皮功能紊乱和凝血相关因子含量的改变，这可能是TWEAK参与动脉粥样硬化早期血管内皮损害的机制。

Aim Using animal model to determine the effect of recombinant human tumor necrosis factor-like weak inducer of apoptosis （rhTWEAK） on aortic endothelial diastolic function and tissue factor and its inhibitor. Methods C57BL mice were randomly divided into four groups ： （ 1 ） rhTWEAK group ： given rhTWEAK by intraperitoneal injection ; （2） rhTWEAK ＋ anti-rhTWEAK group ： after intraperitoneal injection with anti-TWEAK factor, then treated with rhTWEAK factor 15 minutes later; （3）IgG group： treated with nonspecific IgG; （4）Control group： given the same amount 0. 3 mL of normal saline injection. After 7 weeks, we took eyeball blood to separate supernatant and thoracic aortic area to detect the vasodilatation function. Circulating endothelial nitric oxide synthase （eNOS）, high-sensitivity C-reactive pr-otein （hs-CRP）, nitric oxide （NO） and tissue factor （TF） and tissue factor pathway inhibitor （TFPI） were detected in mouse plasma by enzyme linked immunosorbent serologic assay. Results （ 1 ） We found that endothelial diastolic function and the concentration of NO and eNOS in plasma in TWEAK group obviously decreased, TF levels increased sig nificantly and the TFPI decreased when compared with control rhTWEAK ＋ anti-rhTWEAK and IgG group, those factors had significant difference （ P 〈 0. 05 ）, however the hs-CRP content had no obvious change （ P 〉 0. 05 ）. （2） Our results show that the endothelial diastolic function and the content of NO eNOS TF and TFPI and hs-CRP have no obvious differ- ence in the control rhTWEAK ＋ anti-rhTWEAK and IgG group （ P 〉 0. 05）. Conclusion rhTWEAK can induce endo- thelial dysfunction and blood coagulation factor content changes, and this may be a mechanism of vascular endothelial dam- age in early stage atherosclerosis.