Page 57 - 南京医科大学自然版

第46卷第10期 刘怡珂，李超凡，宋海燕，等. 鞣花酸靶向miR⁃18a/TSP⁃1轴减轻缺血性脑损伤［J］.
2026年10月 南京医科大学学报（自然科学版），2026，46（10）：1479-1491 ·1487 ·

A B
miR⁃18a
miRWalk TargetScan
miR⁃19a

miR⁃320
log2FC 12
1 339 77
miR⁃106a 2
1 27
miR⁃146
0
179
miR⁃181 105
-1
miR⁃199a -2
1 207
miR⁃424
StarBase
miR⁃451
MCAO+EA/MCAO 1 2 3

C
GO：0050789（regulation of biological process）
GO：0048518（positive regulation of biological process）
GO：0048511（rhythmic process）
GO：0048519（negative regulation of biological process）
GO：0032501（multicellular organismal process）
GO：0032502（developmental process）
GO：0009987（cellular process）
GO：0050896（response to stimulus）
GO：0022414（reproductive process）
GO：0002376（immune system process）
0 1 2 3 4 5
-lgP F NC miR⁃18a mimic
D
Thbs1 3′⁃UTR⁃WT 5′ ... AACCAGCCUGGCAUUGCACCUUC... 3′ TSP⁃1 180 kDa
rno⁃miR⁃18a⁃5p 3′ ...GAUAGACGUGAUCAUCGUGGAAU... 5′
Thbs1 3′⁃UTR⁃MUT 5′ ... AACCAGCCUGGCAUUACGGUAAG... 3′ β⁃actin 42 kDa

E G H
1.5 ** 1.5 **
1.5
Relative expression level of miR⁃18a 1.0 protein expression TSP⁃1 1.0 Relative luciferase activity 1.0
**

0.5
0.5
0.5

0
Control 50 μmol/L EA 0 NC miR⁃18a mimic 0 WT⁃NC WT⁃miR⁃ MUT⁃NC MUT⁃miR⁃
18a mimic 18a mimic
A：Heatmap of differentially expressed miRNAs in the ischemic penumbra of MCAO rats following EA pretreatment. Columns 1-3 represent
three independent biological replicate measurements，with expression changes presented as log 2FC between the EA+MCAO and MCAO groups. Red
indicates relative upregulation and green indicates relative downregulation ，with color intensity reflecting the magnitude of log 2FC. Differentially
expressed miRNAs were identified using the criteria FC>2.0 or FC<0.5 and P < 0.05（n=3）. B：Venn diagram illustrating the intersection of predicted
miR⁃18a target genes from miRWalk，TargetScan，and StarBase databases. C：GO enrichment analysis of the predicted target genes of miR⁃18a. D：The
predicted binding site of miR⁃18a⁃5p within the Thbs1 3′⁃UTR. The solid lines indicate base pairing，and the dotted lines indicate the mutated binding
site. E：Relative expression levels of miR⁃18a in the OGD model treated with EA，measured by qRT⁃PCR. F，G：Western blot analysis showing the
effect of miR⁃18a mimic on TSP⁃1 protein expression in RBMEC. H：Dual⁃luciferase reporter assay confirming direct targeting：relative luciferase activity
**
demonstrating that miR⁃18a binds to the WT but not the MUT Thbs1 3′⁃UTR. P < 0.01（n=3）.
图4 EA通过上调miR⁃18a并抑制TSP⁃1发挥神经保护作用
Figure 4 EA⁃mediated neuroprotection involves miR⁃18a upregulation and subsequent TSP⁃1 suppression