抗tau蛋白抗体基因疗法对慢性颅脑创伤的治疗作用研究  

Study on the therapeutic effect of anti-tau antibody gene therapy on chronic traumatic encephalopathy

作　　者：柴慈婧 涂悦 张启财 侯伊玲 Chai Cijing;Tu Yue;Zhang Qicai;Hou Yiling(Physical Examination Center of Armed Police Medical Center,Tianjin 300162,China;Institute of Traumatic Brain Trauma and Neurology of Armed Police Medical Center,Tianjin 300162,China)

机构地区：[1]武警特色医学中心体检中心,天津300162 [2]武警特色医学中心颅脑创伤与神经研究所,天津300162

出　　处：《中华神经创伤外科电子杂志》2020年第5期292-298,共7页Chinese Journal Of Neurotraumatic Surgery：Electronic Edition

基　　金：国家自然科学基金(81771350)。

摘　　要：目的探究抗tau蛋白(pTau)基因腺病毒相关病毒(AAV)载体中枢神经系统(CNS)直接干预对慢性颅脑创伤(CTE)的治疗效果。方法随机选取24只C57小鼠按随机数字表法分为3组,正常对照组(sham组)、CTE 4周组和CTE 12周组,每组8只,采用皮质撞击法建立CTE小鼠动物模型,并在实验开始的第4、12周时取脑组织Western blot检测pTau蛋白相对水平评估该模型;将余56只小鼠按随机数字表法分为7组:sham组、noAAV组、AAVrh.10Null组和编码2B6、CisTau、IPN007、PHF1基因载体组,每组8只,除sham组外,各组小鼠通过皮质撞击法构建CTE模型。第3周时sham组与noAAV组注射等量人工脑脊液,其余各组小鼠分别在海马区注射对应基因编码的表达载体,第4周时所有小鼠安乐死,取脑组织进行免疫荧光和Western blot法评估抗pTau抗体表达,并评估对CTE小鼠的治疗效果。结果CTE小鼠模型4和12周组脑组织pTau明显高于sham组,差异均具有统计学意义(P<0.05);AAVrh.10 PHF1与AAVrh.10 IPN治疗组显著降低了小鼠脑组织中pTau的水平,且AAVrh.10 PHF1组优于AAVrh.10 IPN组(P=0.032)。结论抗pTau抗体基因的AAVrh.10病毒载体CNS直接治疗CTE有效,为治疗提供了新的方向。Objective To investigate the direct intervention of anti-tau protein(pTau)gene adeno-associated virus(AAV)vector in the treatment of chronic traumatic encephalopathy(CTE).Methods Twenty-four C57 mice were randomly divided into three groups(n=8),normal control group(sham group),CTE 4 weeks group and CTE 12 weeks group.The CTE mice model was established by cortical impact method.The relative level of pTau protein in brain tissue was detected by Western blot at the 4th and 12th week of the experiment.The remaining 56 mice were randomly divided into 7 groups(n=8):sham group,noAAV group,AAVrh.10Null group and vector group encoding 2B6,CisTau,IPN007 and PHF1 gene vectors.Except for sham group,CTE models were established by cortical impact method.At the 3rd week,the sham group and noAAV group were injected with the same amount of artificial cerebrospinal fluid.The other groups of mice were injected with the corresponding gene encoding expression vector in the hippocampus.At the 4th week,all mice were euthanized.The expression of anti pTau antibody in brain tissue was evaluated by immunofluorescence and Western blot,and the therapeutic effect on CTE mice was also evaluated.Results pTau in CTE mice in the 4 and 12 weeks groups was significantly higher than that in sham group,and the difference were statistically significant(P<0.05).Aavrh.10PHF1 and AAVrh.10IPN treatment group significantly reduced the level of pTau in mouse brain tissue,and aAVRh.10PHF1 group was superior to AAVrh.10IPN group(P=0.032).Conclusion Direct treatment of AAVrh.10 virus vector CNS with anti-pTau antibody gene is effective and provides a new direction for the treatment of CTE.

关 键 词：慢性颅脑创伤 中枢神经系统 腺病毒相关病毒 TAU蛋白 

分 类 号：R651.15[医药卫生—外科学]