Department of Neurology, Zhujiang Hospital, Southern Medical University, Guangzhou, PR China.
Second Clinical College, Southern Medical University, Guangzhou, PR China.
Bioengineered. 2021 Dec;12(1):3348-3357. doi: 10.1080/21655979.2021.1947167.
To investigate the effect of miR-149-5p on sphingosine-1-phosphate receptor 2 (S1PR2) expression level and contents of matrix metalloproteinase (MMP-9) and superoxide dismutase (SOD) in the pericytes after acute cerebral ischemia reperfusion in rats, so as to clarify the neuroprotective molecular mechanism induced by miR-149-5p and provide references for the treatment of neurological diseases, 60 male SD rats aged 7-8 weeks were selected and divided randomly into test group (establishing middle cerebral artery occlusion (MCAO) model) and control group (no modeling). Rat pericytes and peripheral cerebral infarction tissues were collected 12 h, 1 d, 3 d, 5 d, and 7 d after MCAO modeling, respectively. The pericytes were identified by immunofluorescence assay (IFA) and transfected with miR-149-5p. Fluorescence quantitative PCR (FQPCR) and Western blot were adopted to detect S1PR2 expression level. The expression of S1PR2 in MCAO model rats was detected by IFA. Immunohistochemistry (IHC) and quantitative real-time PCR (qRT-PCR) were used to detect the changes of MMP9 protein and mRNA levels of SOD1, SOD2, and SOD3 in brain tissue. The results showed that mRNA level and protein expression level of S1PR2 in the test group were higher than those in the control group three days after MCAO modeling ( < 0.05); the expression of S1PR2 increased 12 h after MCAO modeling and returned to the normal level on the 5th day, and the content of MMP9 protein in brain tissue of the test group was significantly lower than that of the control group ( < 0.05); the mRNA levels and SODs activity of SOD1, SOD2, and SOD3 in the test group were higher than those in the control group ( < 0.05). Therefore, miR-149-5p played a neuroprotective role by regulating S1PR2 to change the expression levels of SODS and MMP9.
为了研究微小 RNA-149-5p(miR-149-5p)对大鼠急性脑缺血再灌注后周细胞中鞘氨醇-1-磷酸受体 2(S1PR2)表达水平及基质金属蛋白酶(MMP-9)和超氧化物歧化酶(SOD)含量的影响,阐明 miR-149-5p 诱导的神经保护分子机制,为神经系统疾病的治疗提供参考,选取 7-8 周龄雄性 SD 大鼠 60 只,随机分为实验组(建立大脑中动脉闭塞(MCAO)模型)和对照组(不建模)。分别于 MCAO 建模后 12 h、1 d、3 d、5 d、7 d 收集大鼠周细胞和周围脑梗死组织。采用免疫荧光法(IFA)鉴定周细胞,转染 miR-149-5p。采用荧光定量 PCR(FQPCR)和 Western blot 检测 S1PR2 表达水平。采用 IFA 检测 MCAO 模型大鼠 S1PR2 的表达。免疫组化(IHC)和实时定量 PCR(qRT-PCR)检测脑组织 MMP9 蛋白和 SOD1、SOD2、SOD3mRNA 水平的变化。结果显示,MCAO 建模后 3 d 实验组 S1PR2mRNA 水平和蛋白表达水平均高于对照组( < 0.05);MCAO 建模后 12 h S1PR2 表达升高,第 5 天恢复正常水平,实验组脑组织 MMP9 蛋白含量明显低于对照组( < 0.05);实验组 SOD1、SOD2、SOD3mRNA 水平和 SOD 活性均高于对照组( < 0.05)。因此,miR-149-5p 通过调节 S1PR2 改变 SODS 和 MMP9 的表达水平发挥神经保护作用。