[1]王钰淳,刘京东,陈莎,等.β-石竹烯通过上调miR-183的表达减轻小鼠缺血性中风损伤[J].第三军医大学学报,2020,42(16):1625-1632.
 WANG Yuchun,LIU Jingdong,CHEN Sha,et al.β-caryophyllene alleviates ischemic stroke injury by upregulating neuronal miR-183 expression in mice [J].J Third Mil Med Univ,2020,42(16):1625-1632.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第16期
页码:
1625-1632
栏目:
神经科学
出版日期:
2020-08-30

文章信息/Info

Title:
β-caryophyllene alleviates ischemic stroke injury by upregulating neuronal miR-183 expression in mice 
作者:
王钰淳刘京东陈莎刘道航董志
重庆医科大学药学院生物化学与分子药理学重点实验室;复旦大学药学院
Author(s):
WANG Yuchun LIU Jingdong CHEN Sha LIU Daohang DONG Zhi
Key Laboratory of Biochemistry and Molecular Pharmacology, College of Pharmacy, Chongqing Medical University, Chongqing, 400016; 2College of Pharmacy, Fudan University, Shanghai, 200433, China
 
关键词:
&beta-石竹烯miR-183缺血性中风NF-&kappaB
Keywords:
&beta-caryophyllene miR-183 ischemic stroke nuclear factor-&kappaB
分类号:
R282.71; R743.3; R966
文献标志码:
A
摘要:
目的探讨β-石竹烯(β-caryophyllene, BCP)是否对缺血性中风有保护作用,以及这种保护作用是否与miR-183和NF-κB的表达有关。方法在动物实验中,将90只C57BL/6雄性小鼠(20~25 g)编号后,根据随机数字法分为3组(n=30):假手术组、模型组、BCP(72 mg/kg)组。模型组和BCP(72 mg/kg)预处理组中动脉栓塞1 h后,再灌注24 h,构建小鼠缺血性中风模型。对小鼠进行神经行为学评分,TTC法测量小鼠脑梗死体积,HE染色和Nissl染色切片观察小鼠大脑神经元、炎性细胞和凋亡小体的病理变化,检测miR-183、NF-κB、p-NF-κB、白介素-1β(IL-1β)、白介素-6(IL-6)的表达,双荧光素酶标记法检测miR-183与NF-κB的靶向关系。提取小鼠的原代神经元,分为3组:正常组、糖氧剥夺(oxygen-glucose deprivation,OGD)损伤组、BCP(10 μmol/L)组。OGD损伤组与BCP(10 μmol/L)预处理组神经元糖氧剥夺1 h后,正常培养24 h,构建神经元缺血性中风模型。观察神经元形态和数量的变化,测定细胞损伤率,测定神经元内miR-183、NF-κB和p-NF-κB的表达,检测炎性因子在神经元内的表达。结果动物实验中,与假手术组相比,模型组小鼠神经行为出现障碍,脑梗死体积增大(P<0.05)。脑组织切片HE染色和Nissl染色显示,缺血一侧的神经元等细胞受到严重损伤。同时,miR-183的表达下降(P<0.05),NF-κB被活化成p-NF-κB,炎性因子NF-κB、IL-1β和IL-6表达升高(P<0.05)。与模型组相比,BCP预处理能改善造模后小鼠神经行为障碍,减少脑梗死体积,保护神经元的形态,减少炎性细胞和凋亡小体的数量,增加皮质中的尼氏阳性细胞数量(P<0.05)。BCP预处理上调miR-183的表达,抑制NF-κB的活化,降低炎性因子的表达(P<0.05)。在神经元实验中,OGD后神经元受损严重,而BCP预处理则保护了神经元,降低神经元死亡率(P<0.05)。BCP预处理还上调神经元中miR-183的表达,抑制NF-κB的活化,降低神经元中炎性因子的表达(P<0.05)。结论缺血性中风后,BCP可能通过上调miR-183表达进一步抑制NF-κB的活化,减少炎性因子释放,从而减轻炎症,发挥脑保护作用。
 
Abstract:

ObjectiveTo investigate the protective effect of β-caryophyllene (BCP) against ischemic stroke injury in mice and clarify whether such protective effects are associated with miR-183 and nuclear factor-κB (NF-κB) expression. MethodsNinety male C57BL/6 mice were randomized equally into 3 groups, namely the sham-operated group, ischemia-reperfusion (I/R) model group, and BCP (72 mg/kg) pretreatment group. The mice in the latter 2 groups were subjected to middle cerebral artery occlusion for 60 min followed by reperfusion for 24 h. The neurological deficits and infarct volume of the mice were measured, and the pathological changes in the brain tissues were observed using HE and Nissl staining. The expression of miR-183, NF-κB, p-NF-κB, interleukin-1β (IL-1β), and IL-6 in the brain were detected, and the targeting relationship between miR-183 and NF-κB was determined. In the in vitro experiment, the cortical neurons isolated from the brain of neonatal mice were divided into normal group, oxygen-glucose deprivation (OGD) group, and OGD group with 10 μmol/L BCP pretreatment. Immunofluorescence assay was employed for morphological observation of the neurons, and the cell injury was assessed with lactate dehydrogenase (LDH) leakage assay. The expression levels of miR-183, IL-1β, IL-6 and NF-κB in the neurons were measured. ResultsIn the mouse models of ischemic stroke, BCP pretreatment significantly improved neurologic deficit score, reduced the infarct volume, lessened pathological changes of the brain tissue, and decreased the number of Nissl-positive cells in the cortex (P<0.05). BCP pretreatment significantly upregulated the expression of miR-183 in the brain tissues of the mice (P<0.05). In the in vitro experiment, BCP pretreatment produced obvious protective effect on neurons and significantly lowered neuronal death rate following OGD (P<0.05). BCP treatment significantly upregulated the expression of miR-183, inhibited NF-κB activation and lowered the expressions of inflammatory factors in the neurons (P<0.05). ConclusionBCP protects the brain tissues against ischemic stroke injury by upregulating the expression of miR-183, which inhibits NF-κB activation and lowers the release of inflammatory factors to alleviate inflammation in the brain.

相似文献/References:

[1]刘京东,陈莎,王钰淳,等.β-石竹烯作用于Notch1/NF-κB信号轴对脑缺血再灌注损伤大鼠的改善作用[J].第三军医大学学报,2021,43(03):218.
 LIU Jingdong,CHEN Sha,WANG Yuchun,et al.β-caryophyllene improves cerebral ischemia reperfusion injury in rats via Notch1/NF-κB signal axis[J].J Third Mil Med Univ,2021,43(16):218.

更新日期/Last Update: 2020-08-17