法舒地尔灌胃对帕金森病小鼠的抗炎和抗氧化作用研究  

Anti-inflammatory and anti-oxidative mechanism of fasudil by gastric perfusion on mice with Parkinson’s diseases

作　　者：樊慧杰 柴智 李艳荣 杨婵娟 王新亮 刘建春 宋国斌[2] 肖保国[3] 马存根 FAN Hui-jie;CHAI Zhi;LI Yan-rong;YANG Chan-juan;WANG Xin-liang;LIU Jian-chun;SONG Guo-bin;XIAO Bao-guo;MA Cun-gen(Neurobiology Research Center/Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to TreatMultiple Sclerosis of State Administration of Traditional Chinese Medicine,Shanxi University of Chinese Medicine,Jinzhong,Shanxi 030619,China;Institute of Brain Science,Shanxi Datong University,Datong,Shanxi 037009,China;Institute of Neurology,Huashan Hospital,Fudan University,Shanghai 200025,China)

机构地区：[1]山西中医药大学神经生物学研究中心/国家中医药管理局多发性硬化益气活血重点研究室,山西晋中030619 [2]山西大同大学脑科学研究所,山西大同037009 [3]复旦大学华山医院神经病学研究所,上海200025

出　　处：《军事医学》2021年第6期437-442,共6页Military Medical Sciences

基　　金：国家自然科学基金面上项目(81703978,81473577,81102552);山西省国际科技合作项目(201703D421016);山西省重点研发计划项目(201803D31091,201803D31209);山西省青年拔尖人才支持计划项目(晋组办字[2019]35号文);山西省自然科学基金(201901D111334);山西省回国留学人员科研资助重点项目(2014-7,2017-129);山西省留学人员科技活动项目择优资助项目([2017]19号文);山西省高等学校科技创新计划项目(2019L0724);山西省重点实验室开放课题(SXIDL-2018-04);山西中医药大学中西医结合防治神经系统疾病研究科技创新团队课题(2018TD-012)。

摘　　要：目的研究法舒地尔(fasudil)灌胃对帕金森病(PD)小鼠的治疗作用及其抗炎与抗氧化机制。方法利用1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)建立PD小鼠模型,将C57BL/6雄性小鼠随机分为2组,法舒地尔治疗组给予小鼠灌胃用药,50 mg/(kg·d),持续14 d;PD模型组灌胃等量生理盐水。采用步态、爬杆实验测试小鼠脚掌与跑道接触面积、头转向下的时间(T-turn)和总时间(T-total)等运动功能相关指标;免疫组织化学染色法检测黑质中酪氨酸羟化酶(TH)阳性神经元;酶联免疫吸附法(ELISA)检测脑组织中炎性因子白细胞介素1β(IL-1β)、IL-6及肿瘤坏死因子α(TNF-α)含量;比色法测定脑组织中丙二醛(MDA)、谷胱甘肽(GSH)的含量;蛋白质印迹技术测定脑组织蛋白质过氧化氢酶(CAT)的表达水平。结果与PD模型组比较,法舒地尔治疗组小鼠的脚掌接触面积降低(P<0.05),T-turn、T-total均缩短(P<0.05,P<0.01),黑质TH阳性神经元致密、增多(P<0.01),炎性因子IL-1β、IL-6及TNF-α含量均下降(均P<0.01),MDA含量降低(P<0.01),GSH、CAT含量均升高(均P<0.01)。结论法舒地尔灌胃可改善PD小鼠的活动能力,减轻运动迟缓症状,对PD有明确的治疗作用,其机制可能与减少炎性因子的分泌、降低氧化产物的水平、提升抗氧化因子的水平有关。Objective To explore the therapeutic effect and the inflammatory and oxidative stress mechanism of fasudil by gastric perfusion in the treatment of Parkinson’s Disease(PD)mice.Methods Mice were randomly divided into two groups.Mice in the PD model group and fasudil group were induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)to establish PD models.At the same time,mice in the fasudil group were intragastrically infused with fasudil until the 14 th day.Mice in the PD model group were treated with the same dosage of sodium chloride solution.The foot contact area was evaluated by gait test.T-turn and T-total were evaluated by pole climbing.Tyrosine hydroxylase(TH)neurons in the substantia nigra of the brain were detected by immunohistochemistry.The levels of inflammatory cytokines interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)in the brain were measured by enzyme-linked immunosorbent assay(ELISA).Malonaldehyde(MDA)and glutathione(GSH)were detected using colorimetric method.Catalase(CAT)in the brain was detected by Western blotting.Results Compared with the model group,the foot contact area was decreased(P<0.05),T-turn and T-total were reduced(P<0.05 and P<0.01,respectively),TH+neurons in the substantia nigra of the brain were increased(P<0.01),the levels of inflammatory cytokines IL-1β,IL-6 and TNF-αdecreased(P<0.01,P<0.01,and P<0.01,respectively),so did the levels of MDA(P<0.01),but the levels of GSH and CAT were increased(P<0.01,and P<0.01,respectively)in the fasudil group.Conclusion Fasudil by gastric perfusion can improve the autonomic activity,reduce the symptoms of motor retardation,and has therapeutic effect in PD mice.The therapeutic effect may be related to suppressing the secretion of inflammatory cytokines,reducing the level of oxidation products and improving the level of antioxidant factors.

关 键 词：法舒地尔 帕金森病 炎性因子 氧化应激 

分 类 号：R742.5[医药卫生—神经病学与精神病学] R971.5[医药卫生—临床医学]