[1]刘高雯,段 颖.急性缺血性脑卒中小鼠模型脑组织中miR-21 表达对炎症反应和细胞凋亡的影响及相关机制研究[J].现代检验医学杂志,2023,38(03):86-91+96.[doi:10.3969/j.issn.1671-7414.2023.03.015]
　LIU Gao-wen,DUAN Ying.Effects of miR-21 Expression on Inflammatory Response and Apoptosis in Brain Tissue of Mice with Acute Ischemic Stroke and Related Mechanisms[J].Journal of Modern Laboratory Medicine,2023,38(03):86-91+96.[doi:10.3969/j.issn.1671-7414.2023.03.015]
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急性缺血性脑卒中小鼠模型脑组织中miR-21 表达对炎症反应和细胞凋亡的影响及相关机制研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年03期
页码:
86-91+96
栏目:
论著
出版日期:
2023-05-15

文章信息/Info

Title:
Effects of miR-21 Expression on Inflammatory Response and Apoptosis in Brain Tissue of Mice with Acute Ischemic Stroke and Related Mechanisms
文章编号:
1671-7414（2023）03-086-07
作者:
刘高雯; 段 颖
（咸阳市中心医院重症医学科，陕西咸阳 712000）
Author(s):
LIU Gao-wen;  DUAN Ying
（Department of Critical Care Medicine，Xianyang Central Hospital， Shaanxi Xianyang 712000，China）
关键词:
微小核糖核酸-21; 急性缺血性脑卒中; 炎症反应; 细胞凋亡
分类号:
R743.33；R-332
DOI:
10.3969/j.issn.1671-7414.2023.03.015
文献标志码:
A
摘要:
目的 研究急性缺血性脑卒中（acute ischemic stroke, AIS）小鼠模型脑组织中miR-21 表达对炎症反应和细胞凋亡的影响，并探讨其相关分子机制。方法 选取C57/BL6 小鼠30 只随机均分为假手术组、短暂局灶性大脑中动脉闭塞（tran-sient middle cerebral artery occlusion，tMCAO）模型组（利用线栓法构建短暂局灶性大脑中动脉闭塞小鼠模型）及miR-21 agomir 激动剂预处理组（tMCAO 术后侧脑室注射miR-21 agomir 激动剂）。RT-PCR 法检测小鼠脑组织中miR-21 表达水平；改良神经严重程度评分（modified neurological severityscore，mNSS）评估小鼠神经功能；2,3,5-三苯基氯化四氮唑（2,3,5-Triphenyltetrazolium chloride，TTC）染色法检测小鼠脑梗死体积；酶联免疫吸附法检测炎症因子（TGF-β1，TNF-α，IL-18 和IL-10）水平变化；原位末端标记（terminal-deoxynucleoitidyl transferase mediatednick end labeling，Tunel）法观察小鼠神经细胞凋亡情况；蛋白免疫印迹法检测TLR4/NF-κB/NLRP3 通路相关蛋白及凋亡蛋白（Bcl-2/Bax）表达情况。结果 假手术组﹑ tMCAO 模型组和miR-21 agomir 组小鼠脑组织中miR-21 表达水平分别为0.83±0.17，0.50±0.11 和1.55±0.32，组间差异有统计学意义（F=43.805，P<0.05）。假手术组小鼠神经功能mNSS 评分均为0 分，tMCAO 组mNSS 评分明显高于miR-21 agomir 组（11.30±3.42 vs 5.61±1.73），差异有统计学意义（t=12.784，P<0.05）。假手术组小鼠脑组织未出现梗死现象，tMCAO 组脑梗死体积（62.72%±5.30%）高于miR-21 agomir 组（38.22%±3.31%），差异有统计学意义（t=31.309，P<0.05）。与tMCAO 组比较，miR-21 过表达明显降低促炎因子TNF-α 和IL-18 水平，升高抗炎因子TGF-β1 和IL-10 水平，差异均有统计学意义（t=3.785 ～ 9.693，均P<0.05）。假手术组脑组织皮层细胞凋亡数约为9.40±5.21 个，明显低于 tMCAO 组的75.52±12.81 个，差异有统计学意义（t=38.552，P<0.05）；miR-21 agomir 组细胞凋亡数约为42.41±9.62 个，低于tMCAO 组，差异有统计学意义（t=19.174，P<0.05）。与tMCAO 组比较，miR-21 过表达组小鼠脑组织中TLR4, NF-κB，p-NF-κB 和NLRP3 蛋白表达降低，Bcl-2/Bax 表达升高，差异均有统计学意义（t=2.314 ～ 3.098，均P ＜ 0.05）。结论 miR-21 在急性缺血性脑卒中后小鼠脑组织中表达明显下降，其过表达可以调节脑缺血后促炎因子和抗炎因子之间的平衡，改善皮层细胞凋亡，可能是通过调控TLR4/NF-κB/NLRP3 通路发挥脑损伤保护作用。
Abstract:
Objective To study the effect of miR-21 expression on inflammatory response and apoptosis in the brain tissue of acute ischemic stroke(AIS) mice, and explore its related molecular mechanism. Methods Thirty C57/BL6 mice were randomly divided into sham operation group, transient middle cerebral artery occlusion(tMCAO) model group (a mouse model of transient focal middle cerebral artery occlusion was established by thread embolism method) and miR-21 agonist pretreatment group (miR-21 agonist was injected into cerebral ventricle after tMCAO operation). The expression of miR-21 in mouse brain was detected by RT-PCR，modified neural severity score (mNSS) was used to evaluate the neural function of mice， 2,3,5-Triphenyltetrazolium chloride (TTC) staining was used to detect the volume of cerebral infarction in mice. Detected inflammatory factors (TGF)-β1，TNF-α，IL-18 and IL-10 level change by enzyme-linked immunosorbent assay. Terminaldeoxynucleoitidyl transferase mediated nick end labeling(Tunel) method was used to observe the neuronal apoptosis in mice, and detected TLR4/NF by Western blot- κ Expression of B/NLRP3 pathway related protein and apoptosis protein (Bcl-2/Bax). Results The expression levels of miR-21 in brain tissues of sham operation group, tMCAO model group and miR-21 agomir group were 0.83 ± 0.17，0.50 ± 0.11 and 1.55 ± 0.32, respectively, with significant difference between groups (F=43.805,P<0.05). The mNSS score of neural function in sham operation group was 0，the mNSS score in the tMCAO group was significantly higher than that in the miR-21 agomir group (11.30 ± 3.42 vs 5.61 ± 1.73), with a statistically significant difference (t=12.784, P<0.05). There was no cerebral infarction in sham operation group，the infarct volume in tMCAO group was about 62.72% ± 5.30%, and that in miR-21 agomir group was about 38.22% ± 3.31%，and the difference was statistically significant (t=31.309, P<0.05). Compared with tMCAO group, overexpression of miR-21 significantly reduced proinflammatory factor TNF- α and IL-18 levels, increasing the anti-inflammatory factor TGF-β1 and IL-10 levels, and the differences were statistically significant (t=3.785 ～ 9.693, all P<0.05). The number of apoptotic cortical cells in sham operation group was about 9.40 ± 5.21, which was significantly lower than that in tMCAO group 75.52 ± 12.81，the difference was statistically significant (t=38.552, P<0.05). The number of apoptosis in miR-21 agomir group was about 42.41 ± 9.62, which was lower than that in tMCAO group, the difference was statistically significant (t=19.174, P<0.05). Compared with tMCAO group, TLR4,NF-κB, p-NF-κB and NLRP3 decreased in brain tissue of miR-21 overexpression group, the expression of Bcl-2/Bax increased, and the differences were statistically significant (t=2.314 ～ 3.098, all P<0.05). Conclusion The expression of miR-21 in the brain tissue of mice after acute ischemic stroke decreased significantly. Its overexpression can regulate the balance between proinflammatory factors and anti-inflammatory factors after cerebral ischemia, and improve the apoptosis of cortical cells, possibly by regulating TLR4/ NF- κ B/NLRP3 pathway plays a protective role in brain injury.

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备注/Memo

备注/Memo:
作者简介：刘高雯（1983-），女，研究生，主治医师，研究方向：神经内科学，E-mail：wendygao 1983@163.com。
更新日期/Last Update: 2023-05-15