Gastrodin exerts neuroprotective effect on stroke rats by regulating
miR-155-mediated Notch pathway

doi:10.3760/cma.j.issn.1007-1245.2022.01.001

Abstract

Abstract: Objective　To explore the protective effect of gastrodin on the nerves of stroke rats and its mechanism. Methods　From January 2020 to March 2021, 94 healthy clean SD rats whose body weight were (250±10) g were divided into a sham operation group, a model group, a low-dose gastrodin group (50 mg/kg), a medium-dose gastrodin group (100 mg/kg), and a high-dose gastrodin group (150 mg/kg). The rats' nerve function was scored by Longa; the brain water content was calculated by weighing; 2, 3, 5-triphenyltetrazolium chloride (TTC) staining was used to observe the cerebral infarction; the morphology of neurons in hippocampus was observed by hematoxylin eosin (HE) staining; the activities of Caspase-3 and caspase-9 were determined by colorimetry; real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of microRNA-155 (miR-155); and Western blot (WB) was used to detect the expression of Notch1 and its ligand Jagged1. The measurement data of normal distribution were expressed as (x±s), and were compared between 3 or more than 3 groups by one-way ANOVA and by SNK-q test between two groups. If P<0.05, there is a statistical difference. Results　Compared with those in the sham operation group, the neurological function in the model group was seriously damaged; the brain water content, cerebral infarction rate, and activities of Caspase-3 and caspase-9 were significantly increased; the expression of miR-155 in the brain was significantly increased; and there were no statistical differences in the expressions of Notch1, Jagged1, and Hes1 (all P>0.05). Compared with those in the model group, the neurological function in the low, medium, and high dose gastrodin groups was reduced; the brain water content [(81.52±2.19)%, (80.39±2.24)%, and (79.33±2.05)% vs. (87.44±2.56)%], cerebral infarction rate [(24.66±2.35)%, (20.87±2.06)%, and (15.65±1.54)% vs. (32.15±3.59)%], and activities of Caspase-3 [(2.48±0.31), (2.12±0.27), and (1.43±0.21) vs (3.27±0.42)] and Caspase-9 [(2.05±0.25), (1.76±0.20), and (1.23±0.16) vs. (2.79±0.36)] were significantly decreased, the expression of miR-155 [(2.15±0.18), (1.97±0.16), and (1.63±0.15) vs. (2.64±0.27)] in the brain was significantly decreased, and the expressions of Notch1 [(0.77±0.13), (0.84±0.16), and (0.95±0.18) vs. (0.42±0.09)], Jagged1 [(0.68±0.10), (0.73±0.15), and (0.87±0.16) vs. (0.38±0.06)], and Hes1 [(0.81±0.12), (0.93±0.15), and (1.07±0.17) vs. (0.43±0.07)] were significantly increased. Conclusion　Gastrodin can down-regulate the expression of miR-155 and activate Notch pathway, and protects the nerves of stroke rats.

Key words: Gastrodin, miR-155, Notch pathway, Stroke rats, Neurological function

摘要： 目的　探究天麻素对脑卒中大鼠神经的保护作用及其机制。方法　2020年1月至2021年3月，将体质量为（250±10）g的清洁级健康雄性SD大鼠94只分为假手术组、模型组、天麻素低剂量组（50 mg/kg）、天麻素中剂量组（100 mg/kg）、天麻素高剂量组（150 mg/kg）。对大鼠神经功能进行Longa评分；称量计算大鼠脑含水量；2，3，5三苯基氯化四氮唑（TTC）染色观察大鼠脑梗死情况；取脑组织，苏木素伊红（HE）染色观察海马组织神经元形态；比色法测定半胱氨酰天冬氨酸特异性蛋白酶（cysteinyl aspartate-specific protease，Caspase）-3、Caspase-9活性；实时定量聚合酶链反应（RT-qPCR）检测微小RNA155（microRNA-155，miR-155）表达；蛋白质免疫印迹检测Notch1及其配体Jagged1的表达。符合正态分布的计量资料以（x±s）表示，多组间比较行单因素方差分析，进一步两组间比较行SNK-q检验，P<0.05为差异有统计学意义。结果　相较于假手术组，模型组大鼠神经功能受损严重，脑含水量、脑梗死率以及Caspase-3、Caspase-9活性显著上升，脑内miR-155表达显著升高，Notch1及其配体Jagged1、下游靶基因Hes1的表达差异无统计学意义（均P>0.05）；相较于模型组，天麻素低、中、高剂量组大鼠神经功能受损程度减轻，脑含水量［（81.52±2.19）%、（80.39±2.24）%、（79.33±2.05）%比（87.44±2.56）%］、脑梗死率［（24.66±2.35）%、（20.87±2.06）%、（15.65±1.54）%比（32.15±3.59）%］以及Caspase-3［（2.48±0.31）、（2.12±0.27）、（1.43±0.21）比（3.27±0.42）］、Caspase-9活性［（2.05±0.25）、（1.76±0.20）、（1.23±0.16）比（2.79±0.36）］显著下降，脑内miR-155表达［（2.15±0.18）、（1.97±0.16）、（1.63±0.15）比（2.64±0.27）］显著下降，Notch1［（0.77±0.13）、（0.84±0.16）、（0.95±0.18）比（0.42±0.09）］、Jagged1［（0.68±0.10）、（0.73±0.15）、（0.87±0.16）比（0.38±0.06）］、Hes1的表达［（0.81±0.12）、（0.93±0.15）、（1.07±0.17）比（0.43±0.07）］显著升高。结论　天麻素可下调miR-155的表达，激活Notch通路，从而对脑卒中大鼠神经起到保护作用。

关键词: 天麻素, miR-155, Notch通路, 脑卒中大鼠, 神经功能

Liu Zhen, Zhang Lei, Hu Canfang, Luo Guojun, Tang Dingzhong. Gastrodin exerts neuroprotective effect on stroke rats by regulating miR-155-mediated Notch pathway[J]. International Medicine and Health Guidance News, 2022, 28(1): 2-7.

刘振, 张磊, 胡灿芳, 罗国君, 汤定中. 天麻素通过调节miR-155介导的Notch通路对脑卒中大鼠发挥神经保护作用[J]. 国际医药卫生导报, 2022, 28(1): 2-7.

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Gastrodin exerts neuroprotective effect on stroke rats by regulating miR-155-mediated Notch pathway

天麻素通过调节miR-155介导的Notch通路对脑卒中大鼠发挥神经保护作用

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