迷迭香酸抑制PI3K/Akt/mTOR信号通路促进细胞自噬缓解帕金森的机制研究  

Mechanism of Rosmarinic Acid Inhibiting PI3K/Akt/mTOR Signaling Pathway and Promoting Autophagy to Alleviate Parkinson’s Disease

作　　者：吕润潇 杜莉莉[2] 周凤华[1] 张立新[1] 刘学勇[1] LYU Runxiao;DU Lili;ZHOU Fenghua;ZHANG Lixin;LIU Xueyong(Shengjing Hospital of China Medical University,Shenyang110000,China;College of Basic Medical Science,China Medical University,Shenyang110000,China)

机构地区：[1]中国医科大学附属盛京医院,沈阳110000 [2]中国医科大学基础医学院,沈阳110000

出　　处：《宁夏医科大学学报》2019年第12期1189-1194,共6页Journal of Ningxia Medical University

基　　金：辽宁省自然科学基金计划重点项目(20180540067)

摘　　要：目的探讨迷迭香酸(RA)对1-甲基-4苯基-1,2,3,6四氢吡啶(MPTP)诱导的小鼠帕金森模型细胞自噬的调控作用及可能的分子机制。方法30只C57BL/6小鼠随机分为对照(control)组、模型(MPTP)组、治疗低剂量(MPTP+low RA)组、治疗中剂量(MPTP+middle RA)组、治疗高剂量(MPTP+high RA)组,每组6只;小鼠连续8 d腹腔注射25 mg·kg^-1 MPTP建立帕金森模型,造模后第9天,三组治疗组连续8 d分别腹腔注射1、4和16 mg·kg^-1的RA。通过爬杆实验和悬挂实验对小鼠进行行为学评分;Real-time PCR及Western blot检测各组小鼠脑组织Bcl-2和Beclin-1 mRNA及蛋白表达水平;Western blot检测各组小鼠脑组织LC3Ⅱ/Ⅰ比值及磷脂酰肌醇3-激酶(phosphatidy linositol 3-kinase,PI3K)、蛋白激酶B(protein dinase B,Akt)和哺乳动物雷帕霉素靶蛋白(mammlian target of rapamycin,mTOR)表达及磷酸化水平。结果与control组相比,MPTP组小鼠运动协调能力下降,Bcl-2、Beclin-1 mRNA水平降低,Bcl-2、Beclin-1和LC3Ⅱ/Ⅰ蛋白表达水平降低,PI3K、Akt和mTOR蛋白磷酸化水平升高(P均<0.05);与MPTP组相比,RA治疗组提升小鼠运动协调能力,Bcl-2、Beclin-1 mRNA和蛋白水平升高,LC3Ⅱ/Ⅰ蛋白表达水平上调,PI3K、Akt和mTOR磷酸化水平下调,并且随RA治疗浓度增加变化越显著(P均<0.05)。结论RA剂量依赖性地抑制PI3K/Akt/mTOR通路促进帕金森小鼠脑组织的细胞自噬和抑制细胞凋亡。Objective To investigate the regulation of rosmarinic acid(RA)on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced autophagy in mouse Parkinson's model and its possible molecular mechanism.Methods C57BL/6 mice were randomly divided into control group,model group(MPTP),low-dose group(MPTP+low RA group),middle-dose group(MPTP+middle RA group),and high-dose group(MPTP+high RA group).The Parkinson models were established by intraperitoneal injection of 25 mg·kg^-1 MPTP for 8 consecutive days.On the 9th day after modeling,the three treatment groups were intraperitoneally injected with 1,4,16 mg·kg^-1 RA for 8 consecutive days.The behavioral scores of mice were evaluated by pole test and traction test.Real-time PCR and Western blot were used to detect the expression of Bcl-2 and Beclin-1 mRNA and protein in brain tissue.Western blot was used to detect the ratio of LC3II/I and the expression of PI3K,Akt and mTOR protein and phosphorylation in brain tissue of each group.Results Compared with control group,the motor coordination ability of MPTP group decreased significantly.The levels of Bcl-2 and Beclin-1 mRNA were decreased significantly,and the expression levels of Bcl-2,Beclin-1 and LC3II/I protein were decreased significantly.The phosphorylation levels of PI3K,Akt and mTOR protein were increased significantly(P<0.05).Compared with MPTP group,RA treatment group improved the motor coordination ability of mice.The levels of Bcl-2 and Beclin-1 mRNA and protein were significantly increased,and the level of LC3 II/I protein was significantly increased.The phosphorylation levels of PI3K,Akt and mTOR were decreased significantly with the increase of RA concentration(P<0.05).Conclusion RA dose-dependent inhibition of PI3K/Akt/mTOR pathway promotes autophagy and inhibits apoptosis in brain tissue of Parkinson's mice.

关 键 词：帕金森 迷迭香酸 磷脂酰肌醇3-激酶 蛋白激酶B 雷帕霉素靶蛋白 自噬 小鼠 

分 类 号：R742.5[医药卫生—神经病学与精神病学]