[1]姬亚杰,任向阳,尹变利,等.微RNA-205对帕金森病大鼠脑额叶皮层纹状体多巴胺能神经元凋亡的影响[J].新乡医学院学报,2020,37(11):1001-1006.[doi:10.7683/xxyxyxb.2020.11.001]
　JI Yajie,REN Xiangyang,YIN Bianli,et al.Effect of microRNA-205 on dopaminergic neuron apoptosis in striatum of frontal cortex of rats with Parkinson′s disease[J].Journal of Xinxiang Medical University,2020,37(11):1001-1006.[doi:10.7683/xxyxyxb.2020.11.001]
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微RNA-205对帕金森病大鼠脑额叶皮层纹状体多巴胺能神经元凋亡的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年11
页码:
1001-1006
栏目:
基础研究
出版日期:
2020-11-05

文章信息/Info

Title:
Effect of microRNA-205 on dopaminergic neuron apoptosis in striatum of frontal cortex of rats with Parkinson′s disease
作者:
姬亚杰1; 任向阳1; 尹变利1; 卢　宏2
(1.郑州大学附属洛阳中心医院神经内科,河南　洛阳　471009;2.郑州大学第一临床学院，河南　郑州　450052)
Author(s):
JI Yajie1; REN Xiangyang1; YIN Bianli1; LU Hong2
(1.Department of Neurology,Luoyang Central Hospital Affiliated to Zhengzhou University,Luoyang 471009,Henan Province,China;2.The First Clinical College of Zhengzhou University,Zhengzhou 450052,Henan Province,China)
关键词:
帕金森病; 纹状体; 微RNA-205; 胰岛素样生长因子-1; 蛋白激酶B; 叉头框蛋白O3a; 多巴胺能神经元
Keywords:
Parkinson′s disease; striatum; microRNA-205; insulin-like growth factor-1; protein kinase B; forkhead box protein O3a; dopaminergic neurons
分类号:
R742.5
DOI:
10.7683/xxyxyxb.2020.11.001
文献标志码:
A
摘要:
目的　探讨微RNA-205（miR-205）通过胰岛素样生长因子-1（IGF-1）/蛋白激酶B（Akt）/叉头框蛋白O3a（FoxO3a）通路对帕金森病（PD）大鼠脑额叶皮层纹状体多巴胺能神经元凋亡的调控作用。方法　将30只雄性Sprague Dawley大鼠随机分为干预组、阴性对照（NC）组、假手术组，每组10只。干预组、NC组大鼠经脑立体定位仪注射6-羟多巴胺建立PD模型，假手术组大鼠注射等体积的生理盐水。建模成功后24 h，干预组和NC组大鼠于纹状体内分别注射miR-205双链寡核苷酸抑制剂（miR-205-inhibitor）及miR-205阴性对照试剂（miR-205-inhibitor-NC）2 μL，假手术组大鼠注射等量的磷酸缓冲盐溶液。观察干预1周后大鼠一般情况，比较各组大鼠异常不自主动作（AIM）评分；脱氧核糖核苷末端转移酶介导的缺口末端标记法检测各组大鼠纹状体神经元凋亡情况；实时荧光定量聚合酶链式反应检测各组大鼠纹状体中miR-205及IGF-1、Akt、FoxO3a mRNA表达；Western blot检测各组大鼠纹状体中IGF-1、Akt、磷酸化Akt（p-Akt）、FoxO3a、磷酸化FoxO3a（p-FoxO3a）蛋白表达。结果　干预1周后，假手术组大鼠进食、饮水、活动正常；NC组大鼠进食、进水、活动减少，活动时动作缓慢或姿势异常，部分有易激惹、狂躁、自残行为；干预组大鼠上述症状轻于NC组。NC组、干预组大鼠AIM评分和神经元凋亡率均显著高于假手术组（P<0.001）；干预组大鼠AIM评分和神经元凋亡率显著低于NC组（P<0.001）。NC组、干预组大鼠纹状体中miR-205 mRNA相对表达量显著高于假手术组（P<0.001），干预组大鼠纹状体中miR-205 mRNA相对表达量显著低于NC组；NC组、干预组大鼠纹状体中IGF-1 mRNA相对表达量显著低于假手术组（P<0.001），干预组大鼠纹状体中IGF-1 mRNA相对表达量显著高于NC组（P<0.001）；3组大鼠纹状体中Akt、FoxO3a mRNA相对表达量比较差异无统计学意义（P>0.05）。NC组、干预组大鼠纹状体中IGF-1蛋白相对表达量及p-Akt/Akt、p-FoxO3a/FoxO3a显著低于假手术组（P<0.001）；干预组大鼠纹状体中IGF-1蛋白相对表达量及p-Akt/Akt、p-FoxO3a/FoxO3a显著高于NC组（P<0.001）。结论　通过抑制大鼠脑额叶皮层纹状体中miR-205表达可抑制多巴胺能神经元凋亡，改善PD症状，其机制可能与上调IGF-1表达及p-Akt、p-FoxO3a水平有关。
Abstract:
Objective　To investigate the regulatory effect of microRNA-205 (miR-205) on apoptosis of dopaminergic neurons in striatum of frontal cortex of Parkinson′s disease(PD) rats through insulin-like growth factor-1 (IGF-1)/protein kinase B (Akt)/forkhead box protein O3a (FoxO3a) pathway.Methods　Thirty male Sprague Dawley rats were randomly divided into the intervention group,negative control (NC)group and sham operation group,with 10 cases in each group.The rats in the intervention group and the NC group were injected with 6-hydroxydopamine via a brain stereotactic apparatus to establish PD model,the rats in the sham operation group were injected with an equal volume of normal saline.At 24 h the successful modeling,the rats in the intervention group were injected with 2 μL miR-205 double-stranded oligonucleotide inhibitor in the striatum,the rats in the NC group were injected with 2 μL miR-205-inhibitor-negative control in the striatum,and the rats in the sham operation group were injected with the same amount of phosphate buffer saline.The general situation of the rats in each group after intervention was observed.The abnormal involuntary movement (AIM) scores of rats in each group were compared.Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay was used to detect the apoptosis in striatal neurons of rats.The expressions of miR-205,IGF-1,Akt and FoxO3a mRNA in striatum of rats were detected by real-time fluorescence quantitative polymerase chain reaction.Western blot was used to detect the expression of IGF-1,Akt,phosphorylated Akt (p-Akt),FoxO3a and phosphorylated FoxO3a (p-FoxO3a) protein in striatum of rats.Results　After one week of intervention,the feed,drank and movement of the rats in sham operation group were all normal;the rats in the NC group had less food and water intake,less activity,slow movement or abnormal posture,and some of them had irritable,manic and self mutilation behaviors;compared with NC group,the above symptoms of the rats in the intervention group were relieved.The AIM score and neuron apoptosis rate of the rats in the NC group and the intervention group were significantly higher than those in the sham operation group(P<0.001),the AIM score and neuron apoptosis rate of the rats in the intervention group were significantly lower than those in the NC group (P<0.001).The relative expressions of miR-205 mRNA in striatum of rats in the NC group and the intervention group were significantly higher than that in the sham operation group (P<0.001),the relative expressions of miR-205 mRNA in striatum of rats in the intervention group was significantly lower than that in the NC group (P<0.001)；the relative expressions of IGF-1 mRNA in striatum of rats in the NC group and intervention group were significantly lower than that in the sham operation group (P<0.001),and the relative expression of IGF-1 mRNA in striatum of rats in the intervention group was significantly higher than that in the NC group (P<0.001);there was no difference in Akt and FoxO3a mRNA among the three groups（P>0.05）.The relative expression of IGF-1,p-Akt/Akt and p-FoxO3a/FoxO3a protein in striatum of rats in the NC group and intervention group were significantly lower than those in the sham operation group (P<0.001),and the relative expression of IGF-1,p-Akt/Akt and p-FoxO3a/FoxO3a protein in striatum of rats in the intervention group were higher than those in the NC group(P<0.001).Conclusion　Inhibition of miR-205 expression in striatum of frontal cortex of rats can improve PD symptoms through inhibiting apoptosis of dopaminergic neurons,and the mechanism may be related to up-regulate the IGF-1 expression and p-Akt and p-FoxO3a.

参考文献/References:

［1］　DUDA J,PTSCHKE C,LISS B.Converging roles of ion channels,calcium,metabolic stress,and activity pattern of substantia nigra dopaminergic neurons in health and parkinson′s disease［J］.J Neurochem,2016,139(Suppl 1):156-178.
［2］　石亮亮,刘亚丹,王博,等.光遗传技术对帕金森病大鼠治疗效果研究［J］.新乡医学院学报,2019,36(12):1121-1124,1129.
［3］　FOFFANI G,OBESO J A.A cortical pathogenic theory of parkinson′s disease［J］.Neuron,2018,99(6):1116-1128.
［4］　MARQUES T M,KUIPERIJ H B,BRUINSMA I B,et al.Micro-RNAs in cerebrospinal fluid as potential biomarkers for Darkinson′s disease and multiple system atrophy［J］.Mol Neurobiol,2017,54(10):7736-7745.
［5］　阎雯,蔡玉香.下调miR-205抑制皮质神经元的缺血性损伤［J］.神经解剖学杂志,2018,34(1):60-66.
［6］　哈力达·巴合提汗,姚亚妮,韩雪洁,等.黄芩苷对帕金森病模型鼠大脑皮质及纹状体区缝隙连接蛋白36表达的影响［J］.中华医学杂志,2019,99(3):218-222.
［7］　PAXINOS G.大鼠脑立体定位图谱［M］.3版.北京:人民卫生出版社,2005:42-43.
［8］　梁建庆,何建成.复方地黄颗粒对阴虚动风证帕金森病异动症模型大鼠神经行为学的影响［J］.解放军医学杂志,2018,43(8):652-656.
［9］　DORSEY E R,SHERER T,OKUN M S,et al.The emerging evidence of the parkinson pandemic［J］.J Parkinsons Dis,2018,8(s1):S3-S8.
［10］　MATHEOUD D,CANNON T,VOISIN A,et al.Intestinal infection triggers Darkinson′s disease-like symptoms in Pink1-/- mice［J］.Nature,2019,571(7766):565-569.
［11］　CORTEZ M A,ANFOSSI S,RAMAPRIYAN R,et al.Role of miRNAs in immune responses and immunotherapy in cancer［J］.Genes Chromosomes Cancer,2019,58(4):244-253.
［12］　谢琼睆,蔡成,龚小慧.高体积分数氧暴露下新生早产大鼠肺组织长链非编码RNA转移相关肺腺癌转录本1和核转录相关因子-2表达变化的意义［J］.中华实用儿科临床杂志,2018,33(21):1640-1644.
［13］　杨恩慧.C1型尼曼—匹克病小鼠神经系统中髓鞘形成障碍及microRNA差异性表达研究［D］.新乡:新乡医学院,2017.
［14］　PATIL K S,BASAK I,PAL R,et al.A proteomics approach to investigate miR-153-3p and miR-205-5p targets in neuroblastoma cells［J］.PLoS One,2015,10(12):e0143969.
［15］　SANEYASU T,TSUCHIHASHI T,KITASHIRO A,et al.The IGF-1/Akt/S6 pathway and expressions of glycolytic myosin heavy chain isoforms are upregulated in chicken skeletal muscle during the first week after hatching［J］.Anim Sci J,2017,88(11):1779-1787.
［16］　WANG Y,JIA L,WANG B,et al.MiR-495/IGF-1/AKT signaling as a novel axis is involved in the epithelial-to-mesenchymal transition of oral squamous cell carcinoma［J］.J Oral Maxillofac Surg,2019,77(5):1009-1021.
［17］　MA X,XU W,ZHANG Z,et al.Salvianolic acid B ameliorates cognitive deficits through IGF-1/Akt pathway in rats with vascular dementia［J］.Cell Physiol Biochem,2017,43(4):1381-1391.
［18］　BEYFUSS K,HOOD D A.A systematic review of p53 regulation of oxidative stress in skeletal muscle［J］.Redox Rep,2018,23(1):100-117.
［19］　KUANG W H,DONG Z Q,TIAN L T,et al.IGF-1 defends against chronic-stress induced depression in rat models of chronic unpredictable mild stress through the PI3K/Akt/FoxO3a pathway［J］.Kaohsiung J Med Sci,2018,34(7):370-376.
［20］　郑世茹,白宏英,程宝仓.姜黄素通过IGF-1/Akt/FoxO3a通路保护多巴胺能神经元细胞［J］.中风与神经疾病,2016,33(4):299-302.

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更新日期/Last Update: 2020-11-05