丁苯酞软胶囊通过下调miR-137促进线粒体自噬对帕金森病大鼠发挥保护作用  

DL-3-n-Butylphthalide soft capsule protects against Parkinson disease in rats by down-regulating miR-137 to promote mitochondrial autophagy

作　　者：王文文[1] 邵彦江[2] 张新乐[1] 张金苹[3] 马琪 徐国卫 WANG Wen-wen;SHAO Yan-jiang;ZHANG Xin-le;ZHANG Jin-ping;MA Qi;XU Guo-wei(Department of Pharmacy,the Seventh People's Hospital of Zhengzhou City,Zhengzhou 450000,China;School of Phar-macy,Zhengzhou University,Zhengzhou 450001,China;Department of Neurology,the Seventh People's Hospital of Zhengzhou City,Zhengzhou 450000,China;Department of Neurology,Zhengzhou Central Hospital Affiliated to Zheng-zhou University,Zhengzhou 450007,China)

机构地区：[1]郑州市第七人民医院药学部,河南郑州450000 [2]郑州大学药学院,河南郑州450001 [3]郑州市第七人民医院神经内科,河南郑州450000 [4]郑州大学附属郑州中心医院神经内科,河南郑州450007

出　　处：《中国病理生理杂志》2021年第12期2172-2179,共8页Chinese Journal of Pathophysiology

基　　金：2015年度河南省医学科技攻关计划项目(No.201504078)。

摘　　要：目的:探究丁苯酞(NBP)软胶囊对帕金森病(PD)大鼠微小RNA-137(miR-137)和线粒体自噬的影响,并探讨其可能的作用机制。方法:SD大鼠随机分为假手术组、模型组、低剂量(36 mg/kg)NBP组、高剂量(72mg/kg)NBP组、NBP(72 mg/kg)+Agomir-NC(miRNA阴性对照;10 nmol)组和NBP(72 mg/kg)+Agomir-137(miR-137模拟物;10 nmol)组,每组18只。采用6-羟基多巴胺(6-OHDA)两点注射法制备PD大鼠模型,假手术组除外;治疗结束后,腹腔注射阿朴吗啡(APO)进行旋转诱导实验,并采用转棒实验评估大鼠的精细运动协调性和平衡能力;免疫组化法检测黑质酪氨酸羟化酶(TH;多巴胺能神经元标志物)和α-突触核蛋白(α-Syn)的阳性表达;透射电子显微镜观察纹状体线粒体自噬情况;JC-1法检测脑黑质-纹状体线粒体膜电位(MMP)变化;RT-qPCR检测大脑miR-137表达水平;Western blot检测线粒体自噬相关蛋白[PTEN诱导假定激酶1(PINK1)、泛素连接酶parkin、微管相关蛋白1轻链3(LC3)和p62]表达。结果:NBP能显著减少PD大鼠的旋转圈数,延长转棒实验掉落潜伏期,增加脑黑质TH的阳性表达,降低α-Syn的阳性表达,降低miR-137表达水平,升高MMP、PINK1和parkin表达及LC3-II/LC3-I比值,并降低p62表达,增强线粒体自噬(P<0.05);而Agomir-137能显著减弱NBP对PD大鼠线粒体自噬的促进作用(P<0.05)。结论:NBP对PD大鼠的保护作用可能与降低miR-137的表达,促进线粒体自噬有关。AIM:To investigate the effects of DL-3-n-butylphthalide(NBP)soft capsule on micro RNA-137(mi R-137)and mitochondrial autophagy in Parkinson disease(PD)rats,and to explore the possible mechanism.METHODS:A total of 108 SD rats were randomly divided into sham group,model group,low-dose(36 mg/kg)NBP group,high-dose(72 mg/kg)NBP group,NBP(72 mg/kg)+Agomir-NC(miRNA negative control;10 nmol)group,and NBP(72 mg/kg)+Agomir-137(miR-137 mimic;10 nmol)group,with 18 in each group.The PD rat model was prepared by two-point injection of 6-hydroxydopamine(6-OHDA).After the treatments,apomorphine(APO)was injected intraperitoneally for rotation induction experiment to evaluate the fine motor coordination and balance ability of the rats.Immunohistochemical staining was used to detect the positive expression of tyrosine hydroxylase(TH;a marker enzyme of dopaminergic neurons in the substantia nigra)andα-synuclein(α-Syn).Transmission electron microscopy was used to observe the autophagy of striatal mitochondria.JC-1 method was used to detect the change of mitochondrial membrane potential(MMP)in the substantia nigra and striatum.RT-qPCR was used to detect the expression level of brain miR-137.Western blot was used to detect the expression of mitochondrial autophagy-related proteins,including PTEN-induced putative kinase 1(PINK1),parkin,microtubule-associated protein 1 light chain 3(LC3)and p62.RESULTS:Treatment with NBP significantly reduced the rotation number of PD rats,and prolonged the fall latency of the rod test.It increased the positive expression of TH and decreased the positive expression ofα-Syn in the substantia nigra.It also reduced the expression level of miR-137 in the brain.It increased the level of MMP,the expression of PINK1 and parkin,and the LC3-II/LC3-I ratio,reduced the expression of p62,and enhanced mitochondrial autophagy(P<0.05).Agomir-137 significantly attenuated the promoting effect of NBP on mitochondrial autophagy in PD rats(P<0.05).CONCLUSION:The protective effect of NBP on PD rats may be related to redu

关 键 词：丁苯酞软胶囊 帕金森病 线粒体自噬 微小RNA-137 

分 类 号：R749.16[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]