法舒地尔减轻新生大鼠脑室周围白质软化的作用机制研究  

Attenuation of periventricular leukomalacia in neonatal rats by fasudil

作　　者：欧阳颖[1] 李健[1] 董红[1] 张羚枚 阮扬皓 唐淑敏 OU-YANG Ying;LI Jian;DONG Hong;ZHANG Ling-mei;RUAN Yang-hao;TANG Shu-min(Department of Paediatrics,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou 510120,China)

机构地区：[1]中山大学孙逸仙纪念医院儿科,广东广州510120

出　　处：《中国病理生理杂志》2020年第10期1796-1802,共7页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.S2013010016308);广州市民生科技攻关计划项目(No.201903010079)。

摘　　要：目的:使用法舒地尔(10 mg/kg)干预缺氧缺血诱导的脑室周围白质软化(periventricular leukomala⁃cia,PVL)新生大鼠模型,探讨法舒地尔是否对大鼠PVL具有治疗作用及其机制。方法:建立缺氧缺血诱导的3日龄新生大鼠PVL模型。将225只3日龄SD大鼠乳鼠,随机分为假手术组、PVL+生理盐水组(PVL组)和PVL+法舒地尔组。每组分为12 h、24 h、48 h、72 h和30 d时点亚组,每个亚组样本量为15只。72 h后取脑组织,制作石蜡切片,HE染色观察脑组织的病理改变;使用RT-qPCR法测ROCK2的mRNA表达水平;Western blot检测ROCK2和NF-κB P65的表达。神经行为学检测和评定于30 d后进行。结果:(1)造模后12、24和48 h PVL组及PVL+法舒地尔组大鼠体重增长速率显著落后于假手术组(P<0.05);(2)HE染色可见假手术组大鼠左右脑室正常,PVL组和PVL+法舒地尔组出现病理形态学变化,而PVL+法舒地尔组相对于PVL组病理改变较轻;(3)PVL组和PVL+法舒地尔组ROCK2的mRNA及蛋白表达较假手术组显著上调(P<0.05);(4)PVL组NF-κB P65的表达在24和48 h较PVL+法舒地尔组和假手术组显著上调(P<0.05)。结论:法舒地尔能够减轻PVL新生大鼠脑部病理损伤,改善远期神经功能,其机制可能与抑制ROCK通路、减少NF-κB P65活化从而减轻炎症损伤有关。AIM:To investigate the effect of fasudil on neonatal rats with periventricular leukomalacia(PVL)and its mechanism.METHODS:Three-day-old neonatal rat PVL model was established by hypoxia and ischemia.The newborn SD rats(3-day-old,n=225)were randomly divided into 3 groups:sham group,PVL group and PVL+fasudil group.The rats in these 3 groups were further divided into 12 h,24 h,48 h,72 h and 30 d subgroups,with 15 neonatal rats each.The neonatal rats in the subgroups were rapidly decapitated and their brains were removed after treatment for 72 h.The pathological changes of brain tissues were observed by HE staining.The expression level of ROCK2 was assessed by RT-qPCR.The protein levels of ROCK2 and NF-κB P65 were analyzed by Western blot.Neurobehavior was observed after 30 d.RESULTS:(1)Growth rates of body weight of the rats in PVL group and PVL+fasudil group were significant⁃ly lower than that in sham group after ischemia(P<0.05).(2)The results of HE staining showed significant pathological changes at 72 h in PVL group and PVL+fasudil group.But the pathological changes in PVL+fasudil group were relatively mild as compared with PVL group.(3)The expression of ROCK2 was significantly increased in PVL group compared with PVL+fasudil group and sham group(P<0.05).(4)The expression of NF-κB P65 was increased in PVL group compared with other groups at 24 h and 48 h(P<0.05).CONCLUSION:Fasudil reduces the pathological damage of brain,and has a long-term neuroprotective effect,which improves neurological prognosis in neonatal rats with PVL by inhibiting ROCK pathway,reducing the activation of NF-κB P65 and attenuating inflammatory damage.

关 键 词：脑室周围白质软化 法舒地尔 Rho/ROCK2信号通路 新生大鼠 NF-κB P65信号通路 

分 类 号：R722.6[医药卫生—儿科] R363.2[医药卫生—临床医学]