Nogo-OMgp/Rho-Rock信号通路在CO中毒急性脑损伤中的作用及盐酸法舒地尔治疗的可行性分析  

Role of neurite outgrowth inhibitor-oligodendrocyte myelin glycoprotein/Ras homologous(Rho)-Rho-associated coiled-coil forming protein kinase signaling pathway in acute brain injury of carbon monoxide poisoning rats and treatment feasibility with hydrochloride fasudil

作　　者：姜文文 毕伟康 李泽坤 王利 王景麟 毕明俊 康海[4] 邹勇[2] 李琴[2] Jiang Wenwen;Bi Weikang;Li Zekun;Wang Li;Wang Jinglin;Bi Mingjun;Kang Hai;Zou Yong;Li Qin(Centre of Integrated Chinese and Western Medicine,School of Basic Medicine,Qingdao University,Qingdao 266071,China;Ward of Integration of Chinese and Western Medicine,Affiliated Yantai Yuhuangding Hospital of Qingdao University,Yantai 264000,China;Clinical School,Weifang Medical College,Weifang 261000,China;Department of Emergency,Affiliated Yantai Yuhuangding Hospital of Qingdao University,Yantai 264000,China)

机构地区：[1]青岛大学基础医学院中西医结合中心,青岛266071 [2]青岛大学附属烟台毓璜顶医院中西医结合病房,烟台264000 [3]潍坊医学院临床学院,潍坊261000 [4]青岛大学附属烟台毓璜顶医院急诊科,烟台264000

出　　处：《中华神经医学杂志》2020年第5期439-448,共10页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金(81571283);山东省重点研发项目(2018GSF118215);山东省自然科学基金(ZR2016HL21、ZR2017LH068);山东省中医药科技发展项目(2015-420、2017-388)。

摘　　要：目的探讨轴突生长抑制因子(Nogo)-少突胶质细胞髓鞘糖蛋白(Omgp)/Ras同源基因(Rho)-Rho相关螺旋卷曲蛋白激酶(Rock)信号通路在一氧化碳(CO)中毒急性脑损伤中的作用,以及Rock抑制剂盐酸法舒地尔对其治疗的可行性。方法将135只健康雄性SD大鼠按随机数字表法分为正常对照组、CO中毒组和盐酸法舒地尔组,每组45只。后2组大鼠采用高压氧舱吸入法建立急性重症CO中毒模型。各组大鼠均接受高压氧治疗2周,其中盐酸法舒地尔组大鼠同时给予腹腔注射盐酸法舒地尔[15 mg/(kg·d),1次/d,共2周],CO中毒组和正常对照组大鼠给予相同体积的生理盐水注射。于造模后1周时应用透射电镜观察各组大鼠海马组织超微结构的改变,于造模后1 d、1周、1个月、2个月时采用免疫组化染色或免疫荧光染色检测各组大鼠脑组织中Nogo、OMgp及Rock阳性细胞染色强度,采用Western blotting检测各组大鼠脑组织中Nogo、OMgp及Rock蛋白的表达水平。结果CO中毒组大鼠海马组织超微结构及血脑屏障损伤明显,以细胞核、线粒体及突触结构改变显著,而盐酸法舒地尔治疗能有效地维持海马组织超微结构及功能的完整性,减轻脑水肿。造模后1 d、1周、1个月、2个月时,CO中毒组大鼠脑组织中Nogo、OMgp、Rock阳性细胞染色强度及Nogo、OMgp、Rock蛋白表达水平均明显高于同一时间点的正常对照组,盐酸法舒地尔组大鼠脑组织中Nogo、OMgp、Rock阳性细胞染色强度及Nogo、OMgp、Rock蛋白表达水平均明显低于同一时间点的CO中毒组,差异均有统计学意义(P<0.05)。结论Nogo-OMgp/Rho-Rock信号通路相关分子的激活与CO中毒急性脑损伤密切相关,而盐酸法舒地尔能有效下调Nogo、OMgp和Rock蛋白的表达水平,从而明显减轻CO中毒后脑损伤程度。Objective To investigate the role of neurite outgrowth inhibitor(Nogo)-oligodendrocyte myelin glycoprotein(Omgp)/Ras homologous(Rho)-Rho-associated coiled-coil forming protein kinase(Rock)signaling pathway in acute brain injury of carbon monoxide(CO)poisoning rats and treatment feasibility with Rho kinase inhibitor hydrochloride fasudil.Methods According to random number table method,135 healthy male SD rats were divided into three groups:a normal control group,a CO poisoning group and a fasudil treatment group(n=45).Rat models of acute severe CO poisoning were established in the CO poisoning group and fasudil treatment group by inhalation method in a hyperbaric oxygen chamber.All rats received hyperbaric oxygen therapy for two weeks.Rats in the farsudil treatment group were intraperitoneally injected with hydrochloride farsudil for intervention(15 mg/[kg·d],once a d for 2 weeks),while those in the CO poisoning and normal control groups received the same volume of normal saline.The ultrastructures of rat brain tissues were observed by transmission electron microscopy one week after modeling.Staining intensities of Nogo-and OMgp-positive cells were detected by immunohistochemistry,and those of Rock-positive cells were analyzed by immunofluorescence one d,one week,one month and two months after modeling.The protein expressions of Nogo,OMgp and Rock in brain tissues were detected by Western blotting one d,one week,one month and two months after modeling.Results In the CO poisoning group,the ultrastructures of brain tissues and blood-brain barrier were damaged obviously,and the changes in nucleus,mitochondria and synaptic structure were obvious;while fasudil treatment could effectively maintain the integrity of ultrastructures and functions of brain tissues,and reduce brain edema.One d,one week,one month and two months after modeling,the staining intensities of Nogo,OMgp and Rock positive cells and protein expression levels of Nogo,OMgp and Rock in the CO poisoning group were significantly higher than those in the n

关 键 词：一氧化碳中毒 急性脑损伤 盐酸法舒地尔 轴突生长抑制因子-少突胶质细胞髓鞘糖蛋白/Ras同源基因-Rho相关螺旋卷曲蛋白激酶信号通路 

分 类 号：R5[医药卫生—内科学] R742[医药卫生—临床医学]