The dysfunction of the blood‐brain barrier (
BBB
) is one of the main pathological features of Alzheimer's disease (
AD
). Memantine (
MEM
), an
N
‐methyl‐
d
‐aspartate (
NMDA
) receptor antagonist, has been reported that been used widely for
AD
therapy. This study was performed to demonstrate the role of the
MEM
in regulating
BBB
permeability in
AD
microenvironment as well as its possible mechanisms. The present study showed that
LINC
00094 was dramatically increased in Abeta
1‐42
‐incubated microvascular endothelial cells (
EC
s) of
BBB
model in vitro. Besides, it was decreased in
MEM
‐incubated
EC
s. Silencing
LINC
00094 significantly decreased
BBB
permeability, meanwhile up‐regulating the expression of
ZO
‐1, occludin and claudin‐5. Furthermore, silencing
LINC
00094 enhance the effect of
MEM
on decreasing
BBB
permeability in
AD
microenvironment. The analysis of the mechanism demonstrated that reduction of
LINC
00094 inhibited Endophilin‐1 expression by up‐regulating miR‐224‐4p/miR‐497‐5p, promoted the expression of
ZO
‐1, occludin and claudin‐5, and ultimately alleviated
BBB
permeability in
AD
microenvironment. Taken together, the present study suggests that the
MEM
/
LINC
00094/miR‐224‐5p (miR‐497‐5p)/Endophilin‐1 axis plays a crucial role in the regulation of
BBB
permeability in
AD
microenvironment. Silencing
LINC
00094 combined with
MEM
provides a novel target for the therapy of
AD
.