Introduction: Fisetin has been demonstrated to inhibit the occurrence of atherosclerosis; however, the mechanism of fisetin suppressing atherosclerosis remains elusive. Methods: The function of fisetin in the inhibition of atherosclerosis was evaluated by hematoxylin and eosin and Oil Red O staining in ApoE−/− mice. Molecular biomarkers of atherosclerosis progression were detected by Western blot and qPCR. Moreover, the inhibition of atherosclerosis on oxidative stress and ferroptosis was evaluated by immunofluorescence staining, qPCR, and Western blot assays. Results: The obtained results showed that serum lipid was attenuated and consequentially the formation of atherosclerosis was also suppressed by fisetin in ApoE−/− mice. Exploration of the mechanism revealed that molecular biomarkers of atherosclerosis were decreased under fisetin treatment. The level of reactive oxygen species and malondialdehyde declined, while the activity of superoxide dismutases and glutathione peroxidase was increased under the fisetin treatment. Additionally, the suppressor of ferroptosis, glutathione peroxidase 4 proteins, was elevated. The ferritin was decreased in the aortic tissues treated with fisetin. Conclusions: In summary, fisetin attenuated the formation of atherosclerosis through the inhibition of oxidative stress and ferroptosis in the aortic tissues of ApoE−/− mice.

1.
La Sala
L
,
Prattichizzo
F
,
Ceriello
A
.
The link between diabetes and atherosclerosis
.
Eur J Prev Cardiol
.
2019
;
26
(
2_Suppl l
):
15
24
.
2.
Bai
T
,
Li
M
,
Liu
Y
,
Qiao
Z
,
Wang
Z
.
Inhibition of ferroptosis alleviates atherosclerosis through attenuating lipid peroxidation and endothelial dysfunction in mouse aortic endothelial cell
.
Free Radic Biol Med
.
2020
;
160
:
92
102
.
3.
Li
J
,
Cao
F
,
Yin
HL
,
Huang
ZJ
,
Lin
ZT
,
Mao
N
, et al
.
Ferroptosis: past, present and future
.
Cell Death Dis
.
2020
;
11
(
2
):
88
.
4.
Vinchi
F
,
Porto
G
,
Simmelbauer
A
,
Altamura
S
,
Passos
ST
,
Garbowski
M
, et al
.
Atherosclerosis is aggravated by iron overload and ameliorated by dietary and pharmacological iron restriction
.
Eur Heart J
.
2020
;
41
(
28
):
2681
95
.
5.
Guo
Z
,
Ran
Q
,
Roberts
LJ
2nd
,
Zhou
L
,
Richardson
A
,
Sharan
C
, et al
.
Suppression of atherogenesis by overexpression of glutathione peroxidase-4 in apolipoprotein E-deficient mice
.
Free Radic Biol Med
.
2008
;
44
(
3
):
343
52
.
6.
Wang
Y
,
Li
L
,
Zhao
W
,
Dou
Y
,
An
H
,
Tao
H
, et al
.
Targeted therapy of atherosclerosis by a broad-spectrum reactive oxygen species scavenging nanoparticle with intrinsic anti-inflammatory activity
.
ACS Nano
.
2018
;
12
(
9
):
8943
60
.
7.
Xiao
L
,
Luo
G
,
Guo
X
,
Jiang
C
,
Zeng
H
,
Zhou
F
, et al
.
Macrophage iron retention aggravates atherosclerosis: evidence for the role of autocrine formation of hepcidin in plaque macrophages
.
Biochim Biophys Acta Mol Cell Biol Lipids
.
2020
;
1865
(
2
):
158531
.
8.
Marques
VB
,
Leal
MAS
,
Mageski
JGA
,
Fidelis
HG
,
Nogueira
BV
,
Vasquez
EC
, et al
.
Chronic iron overload intensifies atherosclerosis in apolipoprotein E deficient mice: role of oxidative stress and endothelial dysfunction
.
Life Sci
.
2019
;
233
:
116702
.
9.
Sampilvanjil
A
,
Karasawa
T
,
Yamada
N
,
Komada
T
,
Higashi
T
,
Baatarjav
C
, et al
.
Cigarette smoke extract induces ferroptosis in vascular smooth muscle cells
.
Am J Physiol Heart Circ Physiol
.
2020
;
318
(
3
):
H508
18
.
10.
Meng
ZJ
,
Liang
HP
,
Zhao
JL
,
Gao
J
,
Liu
CH
,
Ma
XL
, et al
.
HMOX1 upregulation promotes ferroptosis in diabetic atherosclerosis
.
Life Sci
.
2021
;
284
:
119935
.
11.
Khan
N
,
Syed
DN
,
Ahmad
N
,
Mukhtar
H
.
Fisetin: a dietary antioxidant for health promotion
.
Antioxid Redox Signal
.
2013
;
19
(
2
):
151
62
.
12.
Arai
Y
,
Watanabe
S
,
Kimira
M
,
Shimoi
K
,
Mochizuki
R
,
Kinae
N
.
Dietary intakes of flavonols, flavones and isoflavones by Japanese women and the inverse correlation between quercetin intake and plasma LDL cholesterol concentration
.
J Nutr
.
2000
;
130
(
9
):
2243
50
.
13.
Yan
L
,
Jia
QL
,
Cao
H
,
Chen
C
,
Xing
SL
,
Huang
Y
, et al
.
Fisetin ameliorates atherosclerosis by regulating PCSK9 and LOX-1 in apoE(-/-) mice
.
Exp Ther Med
.
2021
;
21
(
1
):
25
.
14.
Althunibat
OY
,
Al Hroob
AM
,
Abukhalil
MH
,
Germoush
MO
,
Bin-Jumah
M
,
Mahmoud
AM
.
Fisetin ameliorates oxidative stress, inflammation and apoptosis in diabetic cardiomyopathy
.
Life Sci
.
2019
;
221
:
83
92
.
15.
Ahmad
S
,
Khan
A
,
Ali
W
,
Jo
MH
,
Park
J
,
Ikram
M
, et al
.
Fisetin rescues the mice brains against D-galactose-induced oxidative stress, neuroinflammation and memory impairment
.
Front Pharmacol
.
2021
;
12
:
612078
.
16.
Li
DL
,
Liu
XM
,
Pi
WH
,
Zhang
Y
,
Yu
L
,
Xu
C
, et al
.
Fisetin attenuates doxorubicin-induced cardiomyopathy in vivo and in vitro by inhibiting ferroptosis through SIRT1/nrf2 signaling pathway activation
.
Front Pharmacol
.
2021
;
12
:
808480
.
17.
Ahmad
S
,
Khan
A
,
Ali
W
,
Jo
MH
,
Park
J
,
Ikram
M
, et al
.
Fisetin rescues the mice brains against D-galactose-induced oxidative stress, neuroinflammation and memory impairment
.
Front Pharmacol
.
2021
;
12
:
612078
.
18.
Li
D
,
Liu
X
,
Pi
W
,
Zhang
Y
,
Yu
L
,
Xu
C
, et al
.
Fisetin attenuates doxorubicin-induced cardiomyopathy in vivo and in vitro by inhibiting ferroptosis through SIRT1/nrf2 signaling pathway activation
.
Front Pharmacol
.
2021
;
12
:
808480
.
19.
Zou
TF
,
Liu
ZG
,
Cao
PC
,
Zheng
SH
,
Guo
WT
,
Wang
TX
, et al
.
Fisetin treatment alleviates kidney injury in mice with diabetes-exacerbated atherosclerosis through inhibiting CD36/fibrosis pathway
.
Acta Pharmacol Sin
.
2023
;
44
(
10
):
2065
74
.
20.
Ikram
M
,
Saeed
K
,
Khan
A
,
Muhammad
T
,
Khan
MS
,
Jo
MG
, et al
.
Natural dietary supplementation of curcumin protects mice brains against ethanol-induced oxidative stress-mediated neurodegeneration and memory impairment via nrf2/TLR4/RAGE signaling
.
Nutrients
.
2019
;
11
(
5
):
1082
.
21.
Zhu
CP
,
Hu
W
,
Wu
H
,
Hu
X
.
No evident dose-response relationship between cellular ROS level and its cytotoxicity: a paradoxical issue in ROS-based cancer therapy
.
Sci Rep
.
2014
;
4
:
5029
.
22.
Mao
GX
,
Zheng
LD
,
Cao
YB
,
Chen
ZM
,
Lv
YD
,
Wang
YZ
, et al
.
Antiaging effect of pine pollen in human diploid fibroblasts and in a mouse model induced by D-galactose
.
Oxid Med Cell Longev
.
2012
;
2012
:
750963
.
23.
Hu
J
,
Jiang
Y
,
Wu
X
,
Wu
Z
,
Qin
J
,
Zhao
Z
, et al
.
Exosomal miR-17-5p from adipose-derived mesenchymal stem cells inhibits abdominal aortic aneurysm by suppressing TXNIP-NLRP3 inflammasome
.
Stem Cell Res Ther
.
2022
;
13
(
1
):
349
.
24.
Moss
JW
,
Ramji
DP
.
Cytokines: roles in atherosclerosis disease progression and potential therapeutic targets
.
Future Med Chem
.
2016
;
8
(
11
):
1317
30
.
25.
Althunibat
OY
,
Al Hroob
AM
,
Abukhalil
MH
,
Germoush
MO
,
Bin-Jumah
M
,
Mahmoud
AM
.
Fisetin ameliorates oxidative stress, inflammation and apoptosis in diabetic cardiomyopathy
.
Life Sci
.
2019
;
221
:
83
92
.
26.
Gaballah
HH
,
El-Horany
HE
,
Helal
DS
.
Mitigative effects of the bioactive flavonol fisetin on high-fat/high-sucrose induced nonalcoholic fatty liver disease in rats
.
J Cell Biochem
.
2019
;
120
(
8
):
12762
74
.
27.
Singh
S
,
Singh
AK
,
Garg
G
,
Rizvi
SI
.
Fisetin as a caloric restriction mimetic protects rat brain against aging induced oxidative stress, apoptosis and neurodegeneration
.
Life Sci
.
2018
;
193
:
171
9
.
28.
Zhang
L
,
Wang
H
,
Zhou
Y
,
Zhu
Y
,
Fei
M
.
Fisetin alleviates oxidative stress after traumatic brain injury via the Nrf2-ARE pathway
.
Neurochem Int
.
2018
;
118
:
304
13
.
29.
Prasath
GS
,
Subramanian
SP
.
Antihyperlipidemic effect of fisetin, a bioflavonoid of strawberries, studied in streptozotocin-induced diabetic rats
.
J Biochem Mol Toxicol
.
2014
;
28
(
10
):
442
9
.
30.
Poston
RN
,
Haskard
DO
,
Coucher
JR
,
Gall
NP
,
Johnson-Tidey
RR
.
Expression of intercellular adhesion molecule-1 in atherosclerotic plaques
.
Am J Pathol
.
1992
;
140
(
3
):
665
73
.
31.
Iiyama
K
,
Hajra
L
,
Iiyama
M
,
Li
H
,
DiChiara
M
,
Medoff
BD
, et al
.
Patterns of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 expression in rabbit and mouse atherosclerotic lesions and at sites predisposed to lesion formation
.
Circ Res
.
1999
;
85
(
2
):
199
207
.
32.
Bourdillon
MC
,
Poston
RN
,
Covacho
C
,
Chignier
E
,
Bricca
G
,
McGregor
JL
.
ICAM-1 deficiency reduces atherosclerotic lesions in double-knockout mice (ApoE[-/-]/ICAM-1[-/-]) fed a fat or a chow diet
.
Arterioscler Thromb Vasc Biol
.
2000
;
20
(
12
):
2630
5
.
33.
Jiang
X
,
Ma
C
,
Gao
Y
,
Cui
H
,
Zheng
Y
,
Li
J
, et al
.
Tongxinluo attenuates atherosclerosis by inhibiting ROS/NLRP3/caspase-1-mediated endothelial cell pyroptosis
.
J Ethnopharmacol
.
2023
;
304
:
116011
.
34.
Khanra
R
,
Dewanjee
S
,
K Dua
T
,
Sahu
R
,
Gangopadhyay
M
,
De Feo
V
, et al
.
Abroma augusta L. (Malvaceae) leaf extract attenuates diabetes induced nephropathy and cardiomyopathy via inhibition of oxidative stress and inflammatory response
.
J Transl Med
.
2015
;
13
:
6
.
35.
Miao
L
,
St Clair
DK
.
Regulation of superoxide dismutase genes: implications in disease
.
Free Radic Biol Med
.
2009
;
47
(
4
):
344
56
.
36.
Zheng
M
,
Li
LZ
,
Liu
YQ
,
Liang
Y
,
Qi
XY
.
Silencing ferritin alleviates atherosclerosis in mice via regulating the expression levels of matrix metalloproteinases and interleukins
.
Acta Biochim Pol
.
2021
;
68
(
4
):
705
10
.
37.
Li
C
,
Zhang
Y
,
Deng
Y
,
Chen
Y
,
Wu
C
,
Zhao
X
, et al
.
Fisetin suppresses ferroptosis through Nrf2 and attenuates intervertebral disc degeneration in rats
.
Eur J Pharmacol
.
2024
;
964
:
176298
.
38.
Takahashi
N
,
Cho
P
,
Selfors
LM
,
Kuiken
HJ
,
Kaul
R
,
Fujiwara
T
, et al
.
3D culture models with CRISPR screens reveal hyperactive NRF2 as a prerequisite for spheroid formation via regulation of proliferation and ferroptosis
.
Mol Cell
.
2020
;
80
(
5
):
828
44.e6
.
39.
Dodson
M
,
Castro-Portuguez
R
,
Zhang
DD
.
NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis
.
Redox Biol
.
2019
;
23
:
101107
.
40.
Osburn
WO
,
Wakabayashi
N
,
Misra
V
,
Nilles
T
,
Biswal
S
,
Trush
MA
, et al
.
Nrf2 regulates an adaptive response protecting against oxidative damage following diquat-mediated formation of superoxide anion
.
Arch Biochem Biophys
.
2006
;
454
(
1
):
7
15
.
41.
Jia
Q
,
Cao
H
,
Shen
D
,
Li
S
,
Yan
L
,
Chen
C
, et al
.
Quercetin protects against atherosclerosis by regulating the expression of PCSK9, CD36, PPARγ, LXRα and ABCA1
.
Int J Mol Med
.
2019
;
44
(
3
):
893
902
.
42.
Cao
H
,
Jia
Q
,
Yan
L
,
Chen
C
,
Xing
S
,
Shen
D
.
Quercetin suppresses the progression of atherosclerosis by regulating MST1-mediated autophagy in ox-LDL-induced RAW264.7 macrophage foam cells
.
Int J Mol Sci
.
2019
;
20
(
23
):
6093
.
43.
Che
J
,
Liang
B
,
Zhang
Y
,
Wang
Y
,
Tang
J
,
Shi
G
.
Kaempferol alleviates ox-LDL-induced apoptosis by up-regulation of autophagy via inhibiting PI3K/Akt/mTOR pathway in human endothelial cells
.
Cardiovasc Pathol
.
2017
;
31
:
57
62
.
44.
Feng
Z
,
Wang
C
,
Yue
J
,
Meng
Q
,
Meng
Q
,
Wu
J
, et al
.
Kaempferol-induced GPER upregulation attenuates atherosclerosis via the PI3K/AKT/Nrf2 pathway
.
Pharm Biol
.
2021
;
59
(
1
):
1106
16
.
You do not currently have access to this content.