TAK-242对心肺复苏后大鼠脑损伤的神经保护作用

TAK-242 confers neuroprotection in rats after cardiopulmonary resuscitation

目的探讨TLR4抑制剂TAK-242对心肺复苏后大鼠脑组织的神经保护作用、可能机制及其临床应用的可能性。方法成年雄性SD大鼠173只，随机分为假手术组（n=20）、复苏组（n=56）、TAK-242低剂量治疗组（n=50）、TAK-242高剂量治疗组（n=47），复苏组和药物治疗组采用窒息方法制作大鼠呼吸心脏骤停模型，模型建立后立即行心肺复苏治疗，药物组心肺复苏治疗的同时通过腹腔注射给予TAK-242治疗。复苏后24h取脑组织标本，免疫印迹法检测NF-κBp65和cleavedcaspase-3的表达，以ELISA法检测大鼠脑组织中炎性因子TNF-α、IL-1β和IL-6的变化，TUNEL法检测神经元的凋亡。结果与假手术组比较，模型组中cleavedcaspase-3、NF-κBp65、TNF-α、IL-1β和IL-6的表达水平显著增加；TAK-242显著降低cleavedcaspase-3、NF-κBp65、IL-6、TNF-α和IL-1β的表达水平，增加NF-κBp65的表达，同时显著降低神经元的凋亡数量，且呈剂量依赖性。结论TAK-242抑制炎性因子的产生对心肺复苏后脑损伤具有一定的保护作用。

Objective To determine the role of TAK-242 on brain injury and neurological out- come after cardiopulmonary resuscitation （CPR） in rats. Methods A total of 173 male Sprague-Daw- ley rats were randomly divided into sham group （n = 20）, CPR group （n = 56）, low-dose TAK-242 treatment group （n = 50） and high-dose TAK-242 treatment group （n = 47）. The animal model of cardiac arrest induced by asphyxia and CPR was established. TUNEL assay was used to determine apoptosis in the cerebral cortex 24 h after recovery of spontaneous circulation. Western blot was used to examine the expression of NF-κB p65 and cleaved caspase-3. Furthermore, a separate set of animals were used to determine the neurologic deficit scales （NDS） at 24, 48 and 72 h post-return of spontaneous circulation. ELISA was used to determine the expression of TNF-α, IL-113 and IL-6 in human cerebrospinal fluid and brain tissues. Results Compared with the sham operation group,the expression levels of cleaved caspase-3 and NF-κB p65, TNF-α, IL-1βand IL-6 in model group were significantly increased, and TAK-242 can significantly decrease expression levels of cleaved caspase-3, NF-κB p65, IL-6, TNF-α, IL-1β, increases the expression of NF-κB p65, while significantly reduces the number of neurons apop- tosis, and it was dose dependent. Conclusion TAK-242 attenuated brain injury and improved neuro- logical outcome after cardiac arrest in rats.