西维来司钠对小鼠颅脑创伤后神经保护作用的研究

目的 探讨中性粒细胞弹性蛋白酶(NE)特异性抑制剂两维来司钠(ONO-5046)在小鼠创伤性颅脑损伤(TBI)后的抗凋亡作用.方法 64只C57BL/6小鼠随机分为对照组、TBI组、TBI+ONO-5046低剂量组(30 mg/kg)以及TBI+ ONO-5046高剂量组(50 mg/kg),所有小鼠均于致伤后24h处死,分别应用实时聚合酶链式反应(PCR)、Westem blot检测挫伤灶周围皮层caspase3、bcl-2、bcl-xl的表达水平.采用末端脱氧核苷酸转移酶介导的生物素脱氧尿嘧啶核苷酸缺口末端标记法( TUNEL)检测皮层神经元凋亡.结果 TBI后损伤灶周围皮层caspase3的mRNA及蛋白表达水平明显升高；bcl-2、bc1-xl的mRNA表达水平有轻度升高；TUNEL阳性细胞显著增多.施加ONO-5046治疗后,caspase3 mRNA及蛋白表达水平明显降低；bcl-2、bcl-xl的mRNA及蛋白表达水平显著升高；TUNEL阳性细胞显著减少,其中50 mg/kg剂量组阳性细胞数减少更为明显(P＜0.05).结论 ONO-5046能够明显抑制TBI后脑组织中caspase3的表达,并且促进bcl-2、bcl-xl的表达,有效抑制神经细胞的凋亡,减轻TBI后的继发性脑损害.

Neuroprotective effect of sivelestat sodium on mice with traumatic brain injury

Objective To investigate the role of neutrophil elastase （NE） specific inhibitor sivelestat sodium （ONO-5046） in regulating neural apoptosis in mouse following traumatic brain injury （TBI）.MethodsA total of 64 C57BL/6 mice were randomly divided into control groups, TBI groups and ONO 5046 treatment groups. The treatment group was divided into two subgroups： high dose group （50 mg/Kg） and low dose group （30 mg/Kg）. All mice were sacrificed at 24 h post-injury. The levels of caspase3, bcl-2 and bcl xl in the brain tissues were detected by real-time polymerase chain reaction （PCR） and Western blot. Furthermore, apoptosis index in the cortical contusion were estimated by trminal deoxynucleotidyl transferase mediated deoxyufidine triphosphate nick end labeling （TUNEL） method.ResultsThe results showed the levels of caspase3 were significantly up-regulated and the levels of bcl 2 and bcl xl were slightly increased. The TUNEL positive cells were significantly increased in the area surrounding the injured brain followed TBI, and the decrease of 50 mg/Kg group was the most obvious （P〈0.05）. Conclusion ONO 5046 administration can inhibit the expression of caspase3, up-regulate the expressions of bcl-2 and bcl-xl, inhibit the apoptosis and reduce the secondary brain injury after TBI.