Nec-1在小鼠脑出血中通过抑制程序性坏死发挥神经保护作用

The role and mechanism of Nec-1 after intracerebral hemorrhage in mice

目的研究程序性坏死的特异性抑制剂Nec-1对小鼠脑出血神经功能的保护作用,并探讨其机制。方法选取健康雄性ICR小鼠,体重25～30 g,采用在纹状体部位注射胶原酶Ⅳ或生理盐水的方法建立小鼠脑出血组或者脑出血对照组,在脑出血前15 min分别在侧脑室注射Nec-1溶液或vehicle溶液。将实验随机分为：对照组、脑出血＋vehicle处理组、脑出血＋Nec-1处理组。分别利用干湿重法测定脑水肿程度,神经功能评分检测神经运动功能,Western blotting检测cleaved caspase-3,Bcl-2蛋白表达情况。结果脑出血加剧脑水肿程度,而Nec-1处理组减轻脑水肿程度（P〈0.05）。经Nec-1处理后,可以提高小鼠脑出血神经运动功能（P〈0.05）。脑出血可以增加cleaved caspase-3蛋白表达,抑制Bcl-2蛋白表达,经Nec-1处理后,抑制cleaved caspase-3的表达水平（P〈0.05）,增加Bcl-2的表达（P〈0.05）。结论 Nec-1对脑出血发挥重要神经保护作用,这种保护作用可能是通过抑制凋亡通路实现的,提示程序性坏死在脑出血中具有重要作用,将为脑出血的治疗提供新的思路。

Objective To investigate the influence of Nec-1,a specific necroptosis inhibitor,on neurofunction after intracerebral hemorrhage and underlying mechanism. Methods Male ICR mice（ 25 ～ 30g） received an infusion of collagenase type IV to induce ICH（ intracerebral hemorrhage） or saline as control into the left striatum. Nec-1 and vehicle were pretreated with a single intracerebroventricular（ i. c. v.） injection into the ipsilateral ventricle 15 min before ICH,respectively. Those mice were randomizes into three groups,control group,ICH and vehicle group,ICH and Nec-1 group. Then brain water content and neurological test were assessed. Meanwhile,the protein of cleaved caspase-3 and Bcl-2 were detected by Western blotting. Results Nec-1-treated group attenuated the brain water content and improved neurological function compared with vehicle group after ICH. Nec-1-treated group inhibited the cleaved caspase-3 and enhanced the Bcl-2protein expression compared with vehicle group after ICH. Conclusion Nec-1 plays neuroprotection role via inhibition of apoptosis after ICH. Meanwhile,it shows that necroptosis has an important role after ICH,which also provides a new way for treating ICH.