川芎提取物通过miR-23a-3p/SNCA轴对帕金森综合征模型MPP^(+)诱导的SH-SY5Y细胞的影响  

Effects of Ligusticum Chuanxiong extract on MPP^(+)-induced SH-SY5Y cells in Parkinson’s syndrome model via miR-23a-3p/SNCA axis

作　　者：谢平安[1] 廖辉[2] 张艳敏[2] 杜从斌 陈慧丰[4] XIE Ping’an;LIAO Hui;ZHANG Yanmin;DU Congbin;CHEN Huifeng(Department of Geriatrics,National Clinical Research Center of Acupuncture and Moxibustion,The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300192;Department of Traditional Chinese Medicine,Tianjin Fourth Central Hospital,Tianjin 300142;Department of Neurology,Tianjin Yellow River Hospital,Tianjin 300110;Department of Neurology,Tianjin Medical University General Hospital Airport Hospital,Tianjin 300308,China)

机构地区：[1]天津中医药大学第一附属医院,国家中医针灸临床医学研究中心老年病科,天津300192 [2]天津市第四中心医院中医科,天津300142 [3]天津市黄河医院神经内科,天津300110 [4]天津医科大学总医院空港医院神经内科,天津300308

出　　处：《西安交通大学学报（医学版）》2023年第4期626-633,共8页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：天津市卫生健康科技项目(No.ZC20230)。

摘　　要：目的探究川芎提取物对MPP^(+)诱导的SH-SY5Y细胞和帕金森综合征的影响。方法1-甲基-4苯基吡啶离子(MPP^(+))干预SH-SY5Y建立帕金森综合征细胞模型(SH-SY5Y-MPP^(+)),川芎提取物干预后检测细胞增殖、凋亡以及miR-23a-3p、SNCA的表达情况。另观察调控miR-23a-3p、SNCA表达后SH-SY5Y-MPP^(+)的变化,双荧光素报告酶验证miR-23a-3p与SNCA的关系。结果SH-SY5Y-MPP^(+)的细胞增殖能力明显低于SH-SY5Y,而凋亡率则高于SH-SY5Y(P<0.05)。川芎提取物干预下,SH-SY5Y-MPP^(+)的增殖能力、Bcl-2、SNCA蛋白升高,凋亡率与miR-23a-3p、Bax蛋白降低(P<0.05)。沉默miR-23a-3p与升高SNCA均可以促进SH-SY5Y-MPP^(+)的增殖能力,抑制凋亡;而升高miR-23a-3p与沉默SNCA则反之(P<0.05)。在线靶基因预测网站发现,miR-23a-3p与SNCA存在可结合的互补位点,双荧光素报告酶显示SNCA-wt的萤火活性在转染了miR-23a-3p模拟物序列后受到了明显的抑制(P<0.05)。在升高miR-23a-3p后,SH-SY5Y-MPP^(+)中SNCA蛋白表达降低,而沉默miR-23a-3p则反之(P<0.05)。拯救实验显示,川芎提取物对SH-SY5Y-MPP^(+)的干预效果被升高miR-23a-3p或沉默SNCA完全逆转(P>0.05);升高miR-23a-3p对SH-SY5Y-MPP^(+)产生的影响则升高SNCA逆转(P>0.05)。结论川芎提取物通过调控miR-23a-3p/SNCA轴影响MPP^(+)诱导的SH-SY5Y生物学行为改变,未来可能是治疗帕金森综合征的新方向。Objective To explore the effects of Ligusticum Chuanxiong extract on MPP^(+)-induced SH-SY5Y cell damage and Parkinson’s syndrome.Methods 1-methyl-4phenylpyridine ion(MPP^(+))interfered with SH-SY5Y to establish a cell model of elderly Parkinson’s syndrome(SH-SY5Y-MPP^(+)).After intervention with Ligusticum Chuanxiong extract,cell proliferation and apoptosis as well as miR-23a-3p and SNCA expressions were detected.In addition,the changes of SH-SY5Y-MPP^(+)after regulating the expression of miR-23a-3p and SNCA were observed,and the relationship between miR-23a-3p and SNCA was verified by dual luciferase reporter.Results The cell proliferation capacity of SH-SY5y-MPP^(+)was significantly lower than that of SH-SY5Y,while the apoptosis rate was higher than that of SH-SY5Y(P<0.05).Under the intervention of Ligusticum Chuanxiong extract,the proliferation ability of SH-SY5Y-MPP^(+),and Bcl-2 and SNCA protein increased,the apoptosis rate,miR-23a-3p,and Bax proteins decreased(P<0.05).Both silencing miR-23a-3p and increasing SNCA could promote the proliferation of SH-SY5Y-MPP^(+)and inhibit apoptosis,while increasing miR-23a-3p and silencing SNCA were the opposite(P<0.05).The online target gene prediction website found that miR-23a-3p and SNCA had complementary sites that could bind,and the dual luciferase reporter enzyme showed that the firefly activity of SNCA-wt was significantly inhibited after the miR-23a-3p mimic sequence was transfected(P<0.05).After increasing miR-23a-3p,the expression of SNCA protein in SH-SY5Y-MPP^(+)decreased,while silencing miR-23a-3p was the opposite(P<0.05).Rescue experiments showed that the intervention effect of Ligusticum Chuanxiong extract on SH-SY5Y-MPP^(+)was completely reversed by increasing miR-23a-3p or silent SNCA(P>0.05);the effect of increasing miR-23a-3p on SH-SY5Y-MPP^(+)increased SNCA reversion(P>0.05).Conclusion Ligusticum Chuanxiong extract can affect the biological behavior changes of SH-SY5Y induced by MPP^(+)by regulating the miR-23a-3p/SNCA axis,which may be a new dir

关 键 词：川芎提取物 miR-23a-3p SNCA MPP^(+)诱导的SH-SY5Y 帕金森综合征 

分 类 号：R741.05[医药卫生—神经病学与精神病学]