中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2013年
3期
226-230
,共5页
卢国辉%蒋春梅%肖振勇%张世忠
盧國輝%蔣春梅%肖振勇%張世忠
로국휘%장춘매%초진용%장세충
MicroRNA-124%帕金森病%多巴胺能神经元%神经保护
MicroRNA-124%帕金森病%多巴胺能神經元%神經保護
MicroRNA-124%파금삼병%다파알능신경원%신경보호
MicroRNA-124%Parkinson's disease%Dopaminergic neuron%Neuroprotection
目的 研究microRNA(miR)-124对帕金森病模型小鼠中脑多巴胺能神经元的神经保护作用,并探讨其免疫炎性调控机制. 方法 采用小胶质细胞系BV2细胞制备炎性反应的细胞模型,实时荧光定量PCR (qRT-PCR)检测miR-21、miR-124、miR-155、miR-146a、miR-181c、miR-221-3p等中枢炎性相关rniRNAs表达；腹腔内注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)制备帕金森病小鼠模型,尾静脉注射miR-124治疗,免疫组织化学染色观察治疗前后多巴胺能神经元凋亡情况(TH蛋白表达)、小胶质细胞活化情况(Iba1蛋白表达),Western blotting及qRT-PCR检测治疗前后细胞凋亡相关蛋白酶caspase-3、caspase-8的表达情况. 结果 miR-124在BV2细胞中表达量明显高于其他5种miRNAs,差异有统计学意义(P＜0.05),且致炎后表达下调幅度最大；与帕金森病模型鼠相比,miR-124治疗后黑质区TH阳性细胞数明显增多,而Iba1阳性细胞数明显减少,caspase-3、caspase-8的表达量亦明显减少,差异有统计学意义(P＜0.05). 结论 miR-124可通过抑制黑质区小胶质细胞活性缓解多巴胺能神经元凋亡进程,可能是帕金森病发病机制中的一个关键因子.
目的 研究microRNA(miR)-124對帕金森病模型小鼠中腦多巴胺能神經元的神經保護作用,併探討其免疫炎性調控機製. 方法 採用小膠質細胞繫BV2細胞製備炎性反應的細胞模型,實時熒光定量PCR (qRT-PCR)檢測miR-21、miR-124、miR-155、miR-146a、miR-181c、miR-221-3p等中樞炎性相關rniRNAs錶達；腹腔內註射1-甲基-4-苯基-1,2,3,6-四氫吡啶(MPTP)製備帕金森病小鼠模型,尾靜脈註射miR-124治療,免疫組織化學染色觀察治療前後多巴胺能神經元凋亡情況(TH蛋白錶達)、小膠質細胞活化情況(Iba1蛋白錶達),Western blotting及qRT-PCR檢測治療前後細胞凋亡相關蛋白酶caspase-3、caspase-8的錶達情況. 結果 miR-124在BV2細胞中錶達量明顯高于其他5種miRNAs,差異有統計學意義(P＜0.05),且緻炎後錶達下調幅度最大；與帕金森病模型鼠相比,miR-124治療後黑質區TH暘性細胞數明顯增多,而Iba1暘性細胞數明顯減少,caspase-3、caspase-8的錶達量亦明顯減少,差異有統計學意義(P＜0.05). 結論 miR-124可通過抑製黑質區小膠質細胞活性緩解多巴胺能神經元凋亡進程,可能是帕金森病髮病機製中的一箇關鍵因子.
목적 연구microRNA(miR)-124대파금삼병모형소서중뇌다파알능신경원적신경보호작용,병탐토기면역염성조공궤제. 방법 채용소효질세포계BV2세포제비염성반응적세포모형,실시형광정량PCR (qRT-PCR)검측miR-21、miR-124、miR-155、miR-146a、miR-181c、miR-221-3p등중추염성상관rniRNAs표체；복강내주사1-갑기-4-분기-1,2,3,6-사경필정(MPTP)제비파금삼병소서모형,미정맥주사miR-124치료,면역조직화학염색관찰치료전후다파알능신경원조망정황(TH단백표체)、소효질세포활화정황(Iba1단백표체),Western blotting급qRT-PCR검측치료전후세포조망상관단백매caspase-3、caspase-8적표체정황. 결과 miR-124재BV2세포중표체량명현고우기타5충miRNAs,차이유통계학의의(P＜0.05),차치염후표체하조폭도최대；여파금삼병모형서상비,miR-124치료후흑질구TH양성세포수명현증다,이Iba1양성세포수명현감소,caspase-3、caspase-8적표체량역명현감소,차이유통계학의의(P＜0.05). 결론 miR-124가통과억제흑질구소효질세포활성완해다파알능신경원조망진정,가능시파금삼병발병궤제중적일개관건인자.
Objective To investigate the neuroprotective effect of microRNA-124 on dopaminergic neurons in Parkinson's disease models and its inflammation-related regulation mechanism.Methods The inflammation cell models were prepared by microglial BV2 murine cells; real-time quantitative PCR (qRT-PCR) was performed to analyze the expressions of inflammation-related miRNAs,including miR-21,miR-124,miR-155,miR-146a,miR-181c and miR-221-3p.The C57BL/6 mouse models of Parkinson's disease were established by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MTPT)intraperitoneal injection,and then,administration of miR-124 via tail vein injection was performed;immunohistochemistry was performed to observe the apoptosis of dopaminergic neurons (TH-staining)and the activation ofmicroglial cells (Iba1-staining) in the substantia nigra of animal models before and after treatment; additionally,Western blotting and qRT-PCR were performed to analyze the expressions of the apoptosis-related proteins (caspase-3 and caspase-8).Results As compared with other 5 miRNAs,miR-124 showed significantly higher expression in BV2 cells (P＜0.05),and presented higher down-regulation after the induction of inflammatory.As compared with those in the Parkinson's disease models,significantly increased TH-positive cells,decreased Iba1-positive cells and down-regulated expressions of caspase-3 and caspase-8 in the substantia nigra of animal models after miR-124 ministration were observed (P＜0.05).Conclusion MiR-124 can slow down the apoptosis of dopaminergic neurons though inhibiting the activation of microglial cells,and maybe it is the key molecule ofpathogenesis of Parkinson's disease.
