HET0016在小鼠纹状体脑出血损伤中的作用及其可能机制

Role of HET0016 in mice with collagenase-induced intracerebral hemorrhage

目的研究20-羟二十烷四烯酸(20-HETE)抑制剂HET0016对脑缺血的作用及其可能机制。方法选择C57品系小鼠,制备胶原酶诱导的纹状体脑出血模型。阻断剂组予以20-HETE的阻断剂HET00162mg/kg,于脑出血模型制备后2h腹腔注射。对照组予以等体积溶剂羟丙基-BETA-环糊精腹腔注射。各组分别于脑出血后各时点取材,测定脑损伤体积,脑含水量的变化,脑出血后神经功能评分,ELISA方法测定脑组织内20-HETE含量,Fluoro Jade B染色标记变性坏死的神经元数量(FJB阳性细胞),免疫荧光染色检测钙结合蛋白(Ibal)和抗髓过氧化物酶(MPO)表达等脑组织小胶质细胞和中性粒细胞炎症反应的情况。结果与对照组相比,阻断剂组脑组织内20-羟二十烷四烯酸表达量均明显降低(N=8,p<0.01),脑损伤体积明显降低(N=5,P<0.05),脑水肿降低(N=8,P<0.05),HET0016明显降低脑出血周围组织内FJB阳性细胞数量(N=5,P<0.01),降低激活的小胶质细胞和浸润的中性粒细胞的数量(N=5,P<0.05)。结论 20-羟二十烷四烯酸在脑出血后加重出血周围神经损伤,其抑制剂可以对脑出血起到治疗作用。

Objective To study the therapeutic role and possible mechanism of 20-HETE inhibitor（N- Hydroxy-N-（4-n-butyl-2-methylphenyl）-formamidine, HET0016）iu intracerebral hemorrhage（ICH）. Methods C57 mice were randomly divided into three groups： ICH group, HET0016 （potently inhibits 20- HETE synthesis by specific cytochrome P450 isoforms ） and vehicle groups. ICH was induced successfully in 12-month-old C57BL/6 mice by intrastriatal injection of collagenase, HETO016 （2 mg/kg） was injected intraperitoneally 2 h after ICH, vehicle-treated mice received the equivalent volume of 10% （2-hydroxypropyl）-β-cyclodextrin in saline. The neurologic function score was determined, and the mice were sacrificed after ICH, brain lesion volume and edema volume were examined, and 20-HETE concentration was determined by ELISA, Fluoro Jade B staining was used to label the death neurons, microglial and neutrophil were stained by immunofluenrescence. Results The concentration of 20-HETE in brain tissue was lower in HET0016-treated group than in ICH or vehicle group 1 day after ICH（n=8, P〈0.01）. HETO016 ameliorated neurologic deficits （n=12/group, P 〈0.05）, and reduced brain lesion volume （n=5, P 〈0.05） and brain edema （n=8, P 〈0.05） and the number of died neurons （n=5, P〈0.O1）, and attenuated microglial activation and ueutrophil infiltration （u=5, both P 〈0.05） 3 days after ICH. Conclusion 20-HETE might be involved in the damage of iutracerebral hemorrhage, its inhibitor plays therapeutic effect on the damage.