ISSN 0006-2979, Biochemistry (Moscow), 2026, Vol. 91, No. 2, pp. 388-403 © The Author(s), 2026. This article is an open access publication.
ISSN 0006-2979, Biochemistry (Moscow), 2026. © The Author(s), 2026. This article is an open access publication.
388
PPARγ Activation Protects against Hydrogen
Peroxide-Induced Oxidative Stress and Apoptosis
in Human Liver Cells
Lingzhi Wu
1,a#
, Fang Chen
2,3,b#
, Kailong Zhong
4,c
, Yunqi An
5,d
,
Yangge Lv
1,e
*, and Xiaofeng Wu
2,3,f
*
1
Department of Pharmacy, the Affiliated Hospital of Jiaxing University (the First Hospital of Jiaxing),
Jiaxing, Zhejiang 314000, China
2
Department of Pharmacy, the First Affiliated Hospital of Xiamen University, Xiamen, Fujian 361000, China
3
Xiamen Key Laboratory for Clinical Efficacy and Evidence-Based Research of Traditional Chinese Medicine,
Xiamen University, Xiamen, Fujian 361015, China
4
Department of Pharmacy, Zhongshan Hospital (Xiamen), Fudan University, Xiamen, Fujian 361015, China
5
Pharmacology and Toxicology, Rutgers University, Piscataway, NJ 08854, USA
a
e-mail: wulingzhi1992@163.com
b
e-mail: fangchen@xmu.edu.cn
c
e-mail: beyondzkl@126.com
d
e-mail: yunqi.an@rutgers.edu
e
e-mail: 00187829@zjxu.edu.cn
f
e-mail: h2009xiaofeng@163.com
Received August 17, 2025
Revised December 30, 2025
Accepted December 30, 2025
Abstract— The deleterious role of oxidative stress in liver damage is a growing problem, and effective
therapeutic interventions are highly warranted. This study evaluated whether peroxisome proliferator-ac-
tivated receptor gamma (PPARγ) activation protects against H
2
O
2
-induced oxidative stress and apoptosis in
human L02 hepatocytes. Cells pretreated with rosiglitazone, a PPARγ agonist, were incubated with H
2
O
2
,
and cell viability was assessed using CCK8 and LDH release assays 24 h after the treatment. The content
of apoptotic cells was determined using Hoechst 33258 staining, and the levels of apoptosis-related pro-
teins were determined by immunoblotting. In addition, several oxidative stress indicators were measured.
Possible involvement of the nuclear factor erythroid 2-related factor (Nrf2) pathway was investigated using
the Nrf2 inhibitor ML385. Rosiglitazone (20 μM) increased cell viability and improved nuclear morpholo-
gy in H
2
O
2
-treated L02 cells, possibly by increasing the Bcl-2/Bax ratio and reducing caspase-3 activation.
Rosiglitazone also decreased reactive oxygen species and malonaldehyde levels, as well as increased the
activities of catalase, glutathione peroxidase, and superoxide dismutase. Rosiglitazone also promoted nu-
clear translocation of Nrf2 and increased the antioxidant levels in H
2
O
2
-treated L02 cells. Inhibition of the
Nrf2 pathway by ML385 partially abolished the rosiglitazone-induced amelioration of oxidative stress and
apoptosis. We conclude that activation of PPARγ protects liver cells against oxidative stress and apoptosis
through the Nrf2 pathway.
DOI: 10.1134/S0006297925602473
Keywords: rosiglitazone, L02 cells, hepatotoxicity, ML385, Nrf2
* To whom correspondence should be addressed.
# These authors contributed equally to this study.
INTRODUCTION
Liver damage is common in chronic conditions
such as viral infections, metabolic diseases, excessive
alcohol and drug consumption, and autoimmune dis-
eases [1, 2]. Both experimental and clinical evidence
indicate that oxidative damage, inflammation, dysreg-
ulation of signaling pathways, and dysfunction of the
innate immune system play critical role in the pro-
gression of liver damage[3]. As the largest detoxifica-
tion organ in the body, the liver metabolizes a wide
ROSIGLITAZONE PROTECTS LIVER CELLS 389
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
range of compounds, a process that generates reac-
tive oxygen species (ROS) as byproducts[4]. Excessive
ROS production disrupts redox balance, ultimately
leading to oxidative stress. Oxidative stress promotes
lipid peroxidation, DNA damage, irreversible protein
modifications, and alterations in signaling pathways
involved in the regulation of gene transcription, pro-
tein expression, and apoptosis, eventually resulting
in hepatic damage [5, 6]. Because of its high meta-
bolic activity, the liver is particularly susceptible to
oxidative stress, which is closely associated with the
emergence and progression of various liver diseases.
Therefore, developing novel therapeutic strategies, in
particular, those targeting oxidative stress-induced
death of hepatocytes, might contribute significantly
to the treatment of liver diseases.
High concentrations of ROS inhibit expressions
of the anti-apoptotic Bcl-2 protein [7]. Excessive ROS
disrupt mitochondrial function, reduce cellular ener-
gy supply, and promote the release of cytochrome c,
resulting in the activation of caspase-3, a key enzyme
of the apoptotic cell pathway [8]. Activated caspase-3
cleaves essential cytoplasmic proteins, leading to DNA
fragmentation in the nucleus, characteristic changes
in the nuclear morphology [9], and ultimately apop-
tosis.
Nuclear transcription factor peroxisome pro-
liferator-activated receptor gamma (PPARγ) plays
a central role in the regulation of glucose and lip-
id metabolism. Its synthetic agonists thiazolidinedi-
ones (TZDs), such as rosiglitazone and pioglitazone,
are a class of oral antihyperglycemic agents used in
clinical practice [10]. Although TZDs possess some
side effects that limit their use for glycemic control,
PPARγ activation has been shown to exert protective
effects in a broad range of other diseases [11, 12].
For instance, rosiglitazone protects against traumatic
brain injury in rats by attenuating neuronal apop-
tosis and autophagy [13]. Pioglitazone increases the
expression of tight junction proteins, improves in-
testinal barrier function, and attenuates dextran
sodium sulfate-induced colitis [14]. Although some
studies have reported that TZDs, particularly trogli-
tazone (first-in-class drug), induce hepatotoxicity by
altering the levels of liver enzymes, the incidence for
such toxicity with rosiglitazone is substantially lower
than that observed with troglitazone[15]. Newer TZD
compounds, such as rosiglitazone and pioglitazone,
have been reported to exhibit milder and reversible
hepatic toxicity profiles [15,16]. The beneficial effects
of rosiglitazone on liver injury have been reported
more often that its adverse effects. For example, pre-
treatment with rosiglitazone inhibited extracellular
signal-related kinase/mitogen-activated protein ki-
nase (ERK/MAPK) signaling and activation of nucle-
ar factor kappa B pathway, thereby effectively pre-
venting the progression of acute liver damage [17].
Rosiglitazone caused 80% higher cell survival in mice
with acute liver injury by reducing liver necrosis and
apoptosis through the modulation of endoplasmic re-
ticulum stress pathways [18]. These findings suggest
that contrary to concerns regarding hepatotoxicity,
recently developed TDZ, such as rosiglitazone, protect
the hepatic functions. Further studies are required to
clarify the underlying mechanisms in order to estab-
lish confidence in their potential therapeutic applica-
tion against clinical hepatotoxicity.
Collectively, previous studies suggest that the
beneficial effects of PPARγ activation are largely at-
tributable to its anti-inflammatory action across a
range of pathological conditions. However, the role of
PPARγ activation in the liver, particularly at the cel-
lular level, has not yet been clarified. In the present
study, we investigated whether rosiglitazone-induced
PPARγ activation protects human L02 hepatocytes
against hydrogen peroxide (H
2
O
2
)-induced oxidative
stress and apoptosis and explored the underlying
molecular mechanism. Our findings may suggest ad-
ditional therapeutic options for the treatment of liver
diseases.
MATERIALS AND METHODS
Materials. Dulbecco’s modified Eagle’s medium
(DMEM) and L02 cells from Jiangsu Kaiji Biotechnology
(Nanjing, China), fetal bovine serum (FBS) from Gibco
Invitrogen (USA), trypsin from Dingguo Changsheng
Biotechnology (Beijing, China), lactate dehydroge-
nase (LDH) assay kit (A020-2-2) from the Jiancheng
Bioengineering Institute (Nanjing, China) were used.
Cell counting kit-8 (CCK-8) (C0037), nucleoprotein ex-
traction kit (P0027), Hoechst 33,258 solution (C1017),
and assay kits for malonaldehyde (MDA) (S0131S),
ROS (S0035S), glutathione peroxidase (GPx) (S0056),
catalase (CAT) (S0051), superoxide dismutase (SOD)
(S0101S), oxidized glutathione (GSSG) (S0053), and re-
duced glutathione (GSH) (S0053) were obtained from
the Beyotime Institute of Biotechnology (Shanghai,
China). Rabbit anti-Bax (sc-70408) and anti-Bcl-2 (sc-
7382) antibodies from Santa Cruz Biotechnology (USA);
rabbit antibodies against caspase-3 (9662), Kelch-like
ECH-associated protein 1 (Keap1) (8047), Nrf2 (12721),
quinone oxidoreductase 1 (NQO1) (3187S), and heme
oxygenase-1 (HO-1) (43966) from Cell Signaling Tech-
nology (USA), rabbit anti-β-actin (AP0731), anti-histone
H3(BS40053) antibodies, and secondary antibodies
from Bioworld Technology (USA) were used. Rosigli-
tazone and ML385 were from Sigma (USA).
Cell culture. Cells were grown in DMEM sup-
plemented with 10% FBS and 0.4% streptomycin/pen-
icillin in 5% CO
2
at 37°C in a humidified atmosphere
WU et al.390
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
and used in the experiments after six passages after
reaching ~80% confluency.
CCK-8 assay. The cytotoxicity of different con-
centrations of H
2
O
2
was determined using the CCK-8
assay. For this, L02 cells were seeded into 96-well
plates (1×10
4
cells/well) and incubated for 24 h at
37°C in a humidified atmosphere containing 5% CO
2
.
Next, the cells were washed twice with PBS and pre-
treated with various concentrations of H
2
O
2
(0, 200,
400, 600, 800, and 1000 μM) for 24 h after two. CCK-8
solution (10 μL) was added to and the cells were in-
cubated for 1 h, The absorbance at 450 nm was then
measured to calculate the concentration of H
2
O
2
that
inhibited cell growth by 50% (IC50) [19].
Treatments. Hepatocyte injury model was es-
tablished by L02 cell exposure to 600 μM H
2
O
2
.
To investigate the protective effects of rosiglitazone,
H
2
O
2
-treated L02 cells were divided into five groups
and incubated for 24h with the following compounds:
control (vehicle + vehicle); drug control (20 μM ro-
siglitazone + vehicle); H
2
O
2
(vehicle + 600 μM H
2
O
2
);
low-dose rosiglitazone (10 μM rosiglitazone + 600 μM
H
2
O
2
); and high-dose rosiglitazone (20 μM rosiglita-
zone + 600 μM H
2
O
2
). Cells in the low- and high-dose
rosiglitazone groups were pretreated with rosiglita-
zone for 2 h prior to H
2
O
2
exposure. After 24 h of
incubation, the cells were assessed using the CCK-8
and LDH release assays, Hoechst 33258 staining, and
Western blotting.
To investigate the protective mechanism of ro-
siglitazone against H
2
O
2
-induced oxidative stress and
apoptosis,
cells were divided into five groups: (1) ve-
hicle + vehicle; (2) vehicle + 600 μM H
2
O
2
; (3) 5 μM
ML385 + 600 μM H
2
O
2
; (4) 20 μM rosiglitazone +
600 μM H
2
O
2
; and (5) 20 μM rosiglitazone + 5 μM
ML385 + 600 μM H
2
O
2
. Cells in the 5 μM ML385 +
600 μM H
2
O
2
group were pretreated with ML385
for 2 h, followed by incubation with ML385 and
H
2
O
2
for 24 h. Cells in the 20 μM rosiglitazone + 5 μM
ML385 + 600 μM H
2
O
2
group were pretreated with
rosiglitazone and ML385 simultaneously for 2 h and
then co-incubated with H
2
O
2
for 24 h.
LDH release assay. The extent of cell death in
H
2
O
2
-treated cells using the LDH release assay. Cells
were seeded in 96-well plates (3×10
5
cells per well)
and treated as indicated, after which the activity
of LDH released from the cells to the culture me-
dium was determined at 450 nm with a microplate
reader. LDH activity reflected membrane damage and
H
2
O
2
-induced cytotoxicity. The same cell density per
well was used for subsequent experiments conducted
in 6-well plates.
Hoechst 33258 staining was used to investigate
whether rosiglitazone protected L02 cells against
H
2
O
2
-induced apoptosis. Cells treated with rosiglita-
zone and H
2
O
2
were washed three times with PBS,
incubated in 0.5 mL of fixation buffer at 37°C for
10 min, washed three times with PBS, and stained
with Hoechst 33258 at room temperature in the dark
for 5 min, and examined under a fluorescence mi-
croscope [20].
Intracellular ROS and MDA assessment. In-
tracellular ROS levels were determined with a ROS
assay kit based on the fluorescence probe 2,7-dichlo-
rodihydrofluorescein diacetate (DCFH-DA) that is oxi-
dized by ROS to fluorescent dichlorofluorescein (DCF).
Thus, DCF fluorescence signal was used to detect the
ROS content in the cells. After treatment with H
2
O
2
and rosiglitazone, cells were incubated with DCFH-DA
(10 μM in PBS) at 37°C for 30 min. Cells were then
washed three times with PBS, collected, and analyzed
for their fluorescence intensity with a flow cytometer
at the excitation wavelength of 488 nm and emission
wavelength of 535 nm. MDA levels were measured
using a commercial MDA assay kit according to the
manufacturer’s instructions (S0131S, Beyotime, China).
Assessment of oxidative stress indicators. The
content of GSH and GSSG and the activities of CAT,
SOD, and GPx were detected using the correspond-
ing commercial kits according to the manufacturers’
instructions.
Western blotting. Expression of Bcl-2 and Bax
and activation of caspase-3 was evaluated using an-
tibodies against caspase-3 (dilution, 1 : 1000), Bax
(1 : 1000), Bcl-2 (1 : 1000), Keap1 (1 : 1000), Nrf2
(1 : 1000), HO-1 (1 : 1000), NQO1 (1 : 1000), β-actin
(1 : 3000), and histone H3 (1 : 500). Cells were incu-
bated with lysis buffer (180 μL of RIPA buffer con-
taining phenylmethylsulfonyl fluorideat a 9 : 1 ratio)
on ice for 30 min and the lysate was centrifuged at
4°C at 12,000g for 10 min to obtain protein-contain-
ing supernatant. Nuclear extract was prepared using
a nucleoprotein extraction kit. Briefly, the cells were
washed twice with PBS, collected by centrifugation at
room temperature at 1000g for 5 min, and homoge-
nized in an ice-cold hypotonic buffer for 10 min. The
lysate was centrifuged at 4°C at 3000g for 5 min; the
precipitate was washed with the hypotonic buffer
and centrifuged at 4°C at 5000g for 5 min. Finally,
0.2 mL of lysis buffer was added to the precipitate,
and the samples were incubated on ice for 20 min
before centrifugation at 4°C at 15,000g for 5 min. The
obtained supernatant containing nuclear proteins was
stored at −40°C.
After determining protein concentration, pro-
teins were separated electrophoretically and trans-
ferred to polyvinylidene fluoride membranes. Fol-
lowing blocking overnight with 5% fat-free milk, the
membranes were probed with primary antibodies
at 4°C and then incubated with an appropriate sec-
ondary antibody conjugated with horseradish peroxi-
dase (dilution, 1 : 5000) at room temperature for 2 h.
ROSIGLITAZONE PROTECTS LIVER CELLS 391
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 1. PPARγ activation ameliorates H
2
O
2
-induced cytotoxicity in L02 cells. a) Viability of L02 cells after 24-h exposure to
different H
2
O
2
concentrations (0, 200, 400, 600, 800, and 1000μM) as assessed by CCK-8 assay. b and c) Viability of L02cells
following indicated treatments assessed by CCK-8 (b) and LDH release (c) assays. Data are shown as mean ± SEM (n=6);
** p < 0.01, ***p < 0.001 vs. Veh + H
2
O
2
group.
Protein bands were visualized using enhanced che-
miluminescence with a gel imaging system (Tanon
Science & Technology Co., China). Four biological re-
peats were performed for each Western blotting ex-
periment.
Statistical analysis was performed with the
SPSS software (version 20.0; IBM, USA). The results
are expressed as mean ± standard error of mean
(SEM). Differences between the groups were evaluat-
ed using one-way ANOVA, followed by the Dunnett’s
post hoc test for multiple comparisons; p-value<0.05
was considered statistically significant.
RESULTS
PPARγ activation ameliorates H
2
O
2
-induced
cytotoxicity in L02 cells. To determine the working
concentrations of H
2
O
2
, L02cells were exposed to dif-
ferent H
2
O
2
concentrations for 24 h, followed by as-
sessment of their viability using CCK-8 assay (Fig. 1a).
Treatment with 600 μM H
2
O
2
reduced cell viability by
~50%; therefore, this concentration was selected for
establishing the cell damage model. Next, we investi-
gated whether rosiglitazone prevented H
2
O
2
-induced
cytotoxicity in L02 cells using CCK-8 detection and
LDH release assay. As shown in Fig. 1, the viability
of H
2
O
2
-treated L02 cells was significantly decreased
compared to the vehicle treatment group (p < 0.001,
Fig. 1b). Treatment with 10 μM rosiglitazone slightly
increased the viability of H
2
O
2
-treated
cells, but the
effect did not reach the level of statistical significance
(p > 0.05, Fig. 1b). Rosiglitazone at the concentration
of 20μM noticeably mitigated the H
2
O
2
-induced cyto-
toxicity, indicating a dose-dependent protective effect
(p < 0.01, Fig. 1b). H
2
O
2
exposure
markedly increased
the LDH release (p < 0.001), whereas pretreatment
with 20μM rosiglitazone reduced it (p < 0.01, Fig. 1c).
In vehicle-treated cells, rosiglitazone did not exert
any cytotoxic effect (p > 0.05), as assessed by both
CCK-8 and LDH release assays.
PPARγ activation attenuates H
2
O
2
-induced
apoptosis in L02 cell. Hoechst 33258 staining was
used to evaluate the protective effect of rosiglitazone
against H
2
O
2
-induced apoptosis in L02 cells. As shown
in Fig. 2b, H
2
O
2
treatment significantly increased the
number of apoptotic cells compared to vehicle-treat-
ed controls (p < 0.01). L02 cells exposed to 600 μM
H
2
O
2
for 24 h exhibited characteristic apoptotic fea-
tures, including chromatin condensation and nucle-
ar shrinkage, manifested as small, bright blue flu-
orescent dots (Fig. 2a). Pre-incubation with 20 μM
rosiglitazone exerted a protective effect on the cells
(p < 0.05, Fig. 2b). Furthermore, treatment with ro-
siglitazone alone (20 μM) did not induce apoptosis
in vehicle-incubated cells. These results suggest that
PPARγ activation protects against H
2
O
2
-induced cell
apoptosis.
WU et al.392
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 2. PPARγ activation inhibits H
2
O
2
-induced L02cell apoptosis. a)Representative images of apoptotic L02cells stained with
Hoechst 33258. b)Cell apoptosis rate in treatments groups. Data are shown as mean ± SEM (n = 4); *p <0.05, ** p< 0.01,
vs. Veh + H
2
O
2
group.
PPARγ activation increases the Bcl-2/Bax ratio
and inhibits caspase-3 activation in H
2
O
2
-treated
cells. To further investigate whether PPARγ activa-
tion mitigates H
2
O
2
-mediated apoptosis, expression
levels of Bax, Bcl-2, and caspase-3 were assessed by
Western blotting. The results demonstrated that H
2
O
2
significantly reduces the Bcl-2/Bax ratio (p < 0.001,
Fig. 3, a and c) and elevated the level of caspase-3
(p < 0.001, Fig. 3, b and d) in L02 cells. Pretreatment
with 20 μM rosiglitazone increased the of Bcl-2/Bax
ratio (p < 0.05, Fig. 3,a andc). Rosiglitazone at 10 μM
and 20 μM significantly inhibited caspase-3 activation
(p < 0.05 and p < 0.01, respectively) (Fig. 3, b and d).
Rosiglitazone (20 μM) treatment alone had no signif-
icant effect on the expression of these proteins in
vehicle-incubated cells.
PPARγ activation alleviates oxidative stress
and enhances the antioxidative capacity in H
2
O
2
-
treated cells. To evaluate the effect of PPARγ acti-
vation on oxidative stress, we measured the content
of ROS as key indicators of oxidative stress based
on DCF fluorescence signal intensity. Treatment
with H
2
O
2
increased ROS levels (p < 0.01, Fig. 4a)
in L02 cells compared to the vehicle group, while
pretreatment with 20 μM rosiglitazone significant-
ly reduced ROS accumulation in H
2
O
2
treated-cells
(p < 0.05, Fig. 4a). Similarly, the content of MDA (lip-
id peroxidation product) was elevated by oxidative
stress (p < 0.01, Fig. 4b) in L02 cells, and pretreat-
ment with 20 μM rosiglitazone markedly decreased
its content in H
2
O
2
-incubated cells (p < 0.05, Fig. 4b).
Moreover, H
2
O
2
treatment decreased the activities
ROSIGLITAZONE PROTECTS LIVER CELLS 393
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 3. PPARγ activation increases Bcl-2/Bax ratio and inhibits caspase-3 activation in H
2
O
2
-treated cells. a and b) Protein
expression levels of Bax, Bcl-2, procaspase-3, and cleaved caspase-3 as detected by Western blotting. c) Bcl-2/Bax ratio
expressed as percentage of the Bcl-2/Bax in Veh + Veh group. d) Caspase-3 activation evaluated as the ratio of caspase-3
fragment to procaspase-3 and expressed as percentage of this ration in Veh + Veh group. β-actin was used as an internal
control. Data are shown as mean ± SEM (n = 4); *p < 0.05, **p < 0.01, ***p < 0.001 vs. Veh + H
2
O
2
group.
of antioxidant enzymes, such as SOD (p < 0.01, Fig. 4c),
CAT (p < 0.05, Fig. 4d), and GPx (p < 0.01, Fig. 4e), and
increased the GSSG/GSH ratio (p < 0.01, Fig. 4f). Pre-
treatment with 20 μM rosiglitazone restored the pa-
rameters to near-normal levels (SOD, p < 0.05, Fig. 4c;
CAT, p < 0.05, Fig. 4d; GPx, p < 0.05, Fig. 4e; GSSG/GSH
ratio, p < 0.01, Fig. 4f). At the same time, treatment
with 20 μM rosiglitazone alone had no significant ef-
fect on ROS levels, MDA content, antioxidant enzyme
activities, or GSSG/GSH ratio in vehicle-treated cells.
PPARγ activation stimulates Nrf2 signaling and
expression of downstream antioxidant proteins in
H
2
O
2
-treated cells. Western blot was performed to
examine the Nrf2 pathway, which plays a crucial role
in the regulation of cytoprotective responses against
oxidative stress [21]. Under physiological conditions,
Nrf2 binds Keap1, forming a complex that remains in
the cytoplasm, thereby maintaining the pathway in an
inactive state. Our data showed that H
2
O
2
exposure
increased the levels of cytoplasmic Nrf2 (p < 0.01,
Fig. 5, a and c) and Keap1 (p < 0.05, Fig. 5, e and f),
while markedly decreasing the content nuclear Nrf2
(p < 0.05, Fig. 5, b and d) compared to the vehicle
group. Pretreatment with 20 μM rosiglitazone signifi-
cantly reduced cytoplasmic Nrf2 (p < 0.05, Fig. 5, a
and c) and Keap1 (p < 0.05, Fig. 5, e and f) and con-
comitantly increased nuclear Nrf2 levels (p < 0.05,
Fig. 5,b andd). Also, expression of the Nrf2 pathway
target proteins, such as HO-1 and NAD(P)H:quinone
oxidoreductase 1 (NQO1), was assessed [22,23]. Treat-
ment with H
2
O
2
decreased expression of both HO-1
(p < 0.01, Fig. 5, e and g) and NQO1 (p < 0.01, Fig. 5, e
and h), but this effect was reversed by pretreatment
with 20 μM rosiglitazone (p < 0.05 for HO-1 and
p < 0.05 for NQO1). Therefore, PPARγ activation res-
cues the Nrf2 pathway suppressed by H
2
O
2
and further
upregulates expression of downstream target proteins.
PPARγ-mediated protection against oxidative
stress and cell apoptosis is partially abolished by
Nrf2 inhibitor in H
2
O
2
-treated cells. To identify the
signal transduction pathway through which PPARγ
activation protects against H
2
O
2
-induced oxidative
stress and apoptosis, L02 cells were co-incubated with
rosiglitazone and Nrf2 inhibitor ML385 (5 μM) [24].
Coadministration of ML385 and rosiglitazone signifi-
cantly attenuated the rosiglitazone-induced increase
in cell viability (p < 0.05, Fig. 6a) and reversed rosigli-
tazone-mediated suppression of LDH release (p < 0.05,
Fig. 6b) in H
2
O
2
-treated cells. Consistent with these
observations, co-treatment with ML385 and rosiglita-
zone partially abolished rosiglitazone-mediated reduc-
tion of cell apoptosis (p < 0.05, Fig. 6, c and d).
PPARγ-mediated protection against oxidative
stress and cell apoptosis is partially abolished by
Nrf2 inhibitor in H2O2-treated cells. Rosiglitazone-in-
duced reduction in the levels of ROS (p < 0.05, Fig. 7a)
WU et al.394
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 4. PPARγ activation alleviates oxidative stress and enhances antioxidant capacity in H
2
O
2
-treated cells. a) ROS lev-
els in L02 cells expressed as percentage of DCF signal intensity relative to that in the Veh + Veh group. b) MDA levels.
c-d) Activities of SOD (c), CAT (d), and GPx (e). f) GSSG/GSH ratio. Data are shown as mean ± SEM (n = 6); * p < 0.05,
** p < 0.01, vs. Veh + H
2
O
2
group.
and MDA (p < 0.05, Fig. 7b) was reversed by coadmin-
istration of ML385. Moreover, cotreatment with rosigl-
itazone and ML385 attenuated rosiglitazone-enhanced
antioxidant defense in H
2
O
2
-treated cells, as indicated
by the decreased activities of CAT (p < 0.05, Fig. 7d)
and GPx (p < 0.05, Fig. 7e) and elevated GSSG/GSH
ratio (p < 0.05, Fig. 7f). Although ML385 also reduced
rosiglitazone-upregulated SOD activity in H
2
O
2
-treat-
ed cells, the effect was not statistically significant
(p > 0.05, Fig. 7c).
Cytoprotective effect of PPARγ activation
against H
2
O
2
-induced injury is associated with the
activation of Nrf2 pathway in L02 cells. Inhibition
of Nrf2 attenuated rosiglitazone-mediated reduction
of cell apoptosis and improvement of antioxidant ca-
pacity, prompting us to use Western blotting to ex-
amine whether this protective effect was associated
with the Nrf2 pathway. Coadministration of ML385
and rosiglitazone elevated cytoplasmic levels of Nrf2
(p < 0.05, Fig. 8,a andc) and Keap1 (p < 0.05, Fig. 8,e
and f) downregulated by rosiglitazone, while rosigli-
tazone-enhanced nuclear Nrf2 level (p < 0.05, Fig. 8,b
and d) was diminished. In line with the inhibition
of Nrf2 nuclear translocation by ML385, coadmin-
istration of ML385 and rosiglitazone downregulat-
ed HO-1 expression level (p < 0.05, Fig. 8, e and g)
in H
2
O
2
- treated cells compared to cells pretreated
with rosiglitazone alone. Additionally, a trend toward
ROSIGLITAZONE PROTECTS LIVER CELLS 395
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 5. PPARγ activation stimulates Nrf2 signaling and expression of downstream antioxidant proteins in H
2
O
2
-treated cells.
Protein levels of cytoplasmic Nrf2, nuclear Nrf2, Keap1, HO-1, and NQO1 were detected by Western blotting (a, b, and e).
Quantification of cytoplasmic Nrf2 (c), nuclear Nrf2 (d), Keap1 (f), HO-1 (g), and NQO1 (h) is presented as a percentage of
the corresponding values in the Veh + Veh group. β-Actin was used as an internal control for cytoplasmic and total protein,
and histone H3 was used as an internal control for nuclear protein. Data are shown as mean ± SEM (n = 4); * p < 0.05,
** p < 0.01, vs. Veh + H
2
O
2
group.
WU et al.396
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 6. Nrf2 inhibitor partially reverses the protective effect of PPARγ activation against oxidative stress and cell apoptosis
in H
2
O
2
-treated cells. Viability of L02cells following indicated treatments assessed by CCK-8(b) and LDH release(c) assays.
c)Representative microphotographs of apoptotic cells stained by Hoechst 33258. d)Cell apoptosis rate in treatment groups.
Data are shown as mean ± SEM (for CCK-8 and LDH release assay, n = 6; for Hoechst 33258 staining, n = 4). # p < 0.05,
## p < 0.01, ### p < 0.001 vs. Rsg + H
2
O
2
group.
ROSIGLITAZONE PROTECTS LIVER CELLS 397
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 7. PPARγ-mediated protection against oxidative stress and cell apoptosis is partially abolished by Nrf2 inhibitor in
H
2
O
2
-treated cells. a)ROS levels in L02 cells expressed as percentage of DCF signal intensity relative to that in the Veh + Veh
group. b) MDA levels. c-d) Activities of SOD (c), CAT (d), and GPx (e). f) GSSG/GSH ratio. Data are shown as mean ± SEM
(n = 6); # p < 0.05, ## p < 0.01, ### p < 0.001 vs. Rsg + H
2
O
2
group.
reduced NQO1 levels (p > 0.05, Fig. 8, e and h) was
observed in H
2
O
2
-induced cells cotreated with ML385
and rosiglitazone, although this effect did not reach
statistical significance relative to rosiglitazone admin-
istration alone. Collectively, these data indicate that
the Nrf2 signaling pathway plays a critical role in the
protective effect of PPARγ activation against H
2
O
2
-
induced oxidative stress and cell apoptosis.
DISCUSSION
The present study demonstrated that PPARγ ac-
tivation attenuates oxidative stress and apoptosis in
L02 cells through the Nrf2 signaling pathways. Pre-
treatment with rosiglitazone at relatively higher con-
centration significantly enhanced cell viability and
suppressed chromatin condensation in H
2
O
2
-treated
WU et al.398
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 8. Cytoprotective effect of PPARγ activation against H
2
O
2
-induced injury in L02 cells is associated with the activation
of Nrf2 pathway. a, b, and e) Protein levels of cytoplasmic Nrf2, nuclear Nrf2, Keap1, HO-1, and NQO1. Quantification of
cytoplasmic Nrf2 (c), nuclear Nrf2 (d), Keap1 (f), HO-1 (g), and NQO1 (h) expressed as percentage of the corresponding
values in the Veh + Veh group. β-Actin was used as an internal control for cytoplasmic and total protein, and histone
H3 was used as an internal control for nuclear protein. Data are shown as mean ± SEM (n = 6); # p < 0.05, ## p < 0.01,
###
p < 0.001 vs. Rsg + H
2
O
2
group.
ROSIGLITAZONE PROTECTS LIVER CELLS 399
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
L02 cells. Also, rosiglitazone pretreatment markedly
reduced decreased ROS levels and MDA content, while
restoring the activity of antioxidant enzymes, such as
SOD, CAT, and GPx, indicating a significant mitigation
of oxidative stress. Pretreatment with ML385 partial-
ly abolished the protective effects of rosiglitazone on
cell viability and redox homeostasis in H
2
O
2
-exposed
cells. Collectively, these findings suggest that PPARγ
activation protects against oxidative stress and cell
apoptosis, an effect that is strongly associated with
the Nrf2 pathway activation.
The onset and development of liver diseases
may be explained by the development of oxidative
stress, a key factor implicated in the pathogenesis
of numerous hepatic disorders [25, 26]. ROS play an
important role in several cellular functions, such as
signal transduction, defense against harmful stimuli,
and expression of genes involved in cell proliferation
and death. However, excessive ROS accumulation that
overwhelms endogenous antioxidant defense capaci-
ty disrupts redox homeostasis, leading to oxidative
stress and severe cellular damage. H
2
O
2
has a long
lifetime and could easily convert to the highly re-
active and cytotoxic hydroxyl radical [27]. H
2
O
2
is
widely used to induce the oxidative stress in various
cell types [28, 29]. In the present study, exposure of
L02 cells to 600 μM H
2
O
2
resulted in a reduced cell
viability and oxidative stress induction, as evidenced
by increased ROS levels, elevated MDA content, and
suppression of the antioxidant defense system. Previ-
ous studies have reported that rosiglitazone inhibits
inflammatory responses and reduces oxidative stress,
thereby ameliorating renal tubular injury. Similarly,
pioglitazone administration decreased the ROS levels
while upregulating nitric oxide synthase phosphory-
lation and nitric oxide production, protecting aging
cerebral arteries against oxidative stress damage[30].
Consistent with previous studies, our data demon-
strate that administration of 20 μM rosiglitazone re-
versed H
2
O
2
-induced oxidative damage in H
2
O
2
-treat-
ed L02 cells.
ROS are mainly generated in mitochondria; there-
fore, mitochondria are the primary organelles dam-
aged by excessive ROS exposure. Mitochondrial DNA
(mtDNA) is particularly susceptible to oxidative dam-
age due to its unique structural features, lack of his-
tone protection, and close proximity to the mitochon-
drial electron transport chain, a major site of ROS
production [31]. Damage to mtDNA compromises
ATP production and further enhances ROS genera-
tion. There is a growing body of evidence indicating
that excessive mitochondrial ROS accumulation con-
tributes to abnormal mitochondrial fission/fusion and
calcium overload, which, in turn, promotes additional
ROS production[32]. These pathological processes in-
teract and form a vicious cycle that triggers apoptotic
signaling. Alterations in the Bax/Bcl-2 ratio promote
the release of cytochrome c and trigger a caspase
cascade, playing a critical role in mitochondrial dys-
function and oxidative stress-induced apoptosis. As a
potent oxidant, H
2
O
2
induces the permeability transi-
tion pore opening and increases mitochondrial per-
meability, leading to the caspase-dependent apopto-
sis in mammalian cells [33]. Here, we observed that
H
2
O
2
-stimulated L02 cells exhibited chromatin con-
densation (a hallmark of apoptosis), reduced Bcl-2/
Bax ratio, and elevated levels of cleaved caspase-3, a
key executor of cell apoptosis. Previous studies [19]
demonstrated that epigallocatechin-3-gallate alleviates
high-fat diet-induced nonalcoholic fatty liver disease
by inhibiting apoptosis though the ROS/mitogen-ac-
tivated protein kinase pathway [19]. Similarly, mel-
atonin attenuates oxidative stress, inhibits apoptosis
in hepatocytes, and suppresses autophagy in animal
models of acute liver failure [34]. In addition to the
antioxidant effect, PPARγ activation significantly re-
duced H
2
O
2
-induced cell apoptosis, supporting the idea
that antioxidants exhibit protective effects and hold
a strong therapeutic potential against hepatic injury.
The transcription factor Nrf2 is a cellular redox
sensor expressed in multiple organs. Accumulating
evidence indicates that chronic and severe stress can
suppress Nrf2 activity and that dysregulation of the
Nrf2 pathway is involved in the pathogenesis of sev-
eral liver diseases [35]. We found that exposure to
H
2
O
2
elevated the Keap1 level and reduced nuclear
translocation of Nrf2, thereby affecting the down-
stream antioxidant enzymes and reducing the anti-
oxidant capacity of cells. The binding of ligands to
PPARγ activates it, leading to the PPARγ translocation
to the nucleus and interaction with the retinoid X
receptor (RXR) partner protein. The formed complex
binds to PPAR response elements (PPREs) located in
the promoter regions of PPARγ-regulated genes. Sev-
eral studies have demonstrated that PPARγ agonists
enhance the PPARγ transcriptional activity and up-
regulate the expression of key antioxidant proteins,
such as HO-1, CAT, and GPx, thereby counteracting
oxidative stress [36, 37]. Interestingly, a previous
study identified several putative PPREs in the pro-
moter region of Nrf2 gene; however, their function
significance remains unclear, suggesting a potential
direct regulatory association between PPARγ and
Nrf2 [38]. This might partially explain how PPARγ
activation promotes Nrf2 translocation to the nu-
cleus, elevates activities of SOD, CAT, and GPx, and
upregulates HO-1 and NQO1 expression, enhancing
the antioxidant defense. Consequently, rosiglitazone
treatment increased cell viability and reduced apop-
tosis. ML385 inhibited activation of the Nrf2 pathway
and attenuated rosiglitazone-induced upregulation
of antioxidant enzyme activities and expressions.
WU et al.400
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Fig. 9. Proposed mechanism by which rosiglitazone attenuates oxidative stress and apoptosis through activation of the
Nrf2 signaling in L02 cells.
Coadministration of rosiglitazone and ML385 weakened
the protective effect of rosiglitazone against oxidative
stress and cell apoptosis. Intriguingly, several studies
have identified antioxidant response elements (AREs)
in the PPARγ gene promoter region and demonstrated
that increased nuclear translocation of Nrf2 upreg-
ulates PPARγ expression [39, 40]. Nrf2 deletion sig-
nificantly reduced PPARγ expression in vivo [41]. We
propose an existence of a crosstalk between the Nrf2
and PPARγ pathways that creates a positive feedback
loop and promotes expression of downstream antiox-
idant genes. Further studies are required to confirm
the reciprocal regulation between PPARγ and Nrf2
and to clarify the underlying signaling mechanisms.
CONCLUSION
PPARγ activation protects cells against H
2
O
2
-in-
duced oxidative stress through activation of the Nrf2
pathway, as indicated by increased activities of an-
tioxidant enzymes (CAT, SOD, and GPx), upregulated
expression of HO-1 and NQO1, and simultaneous in-
crease in the Bcl-2 level, reduction in the Bax con-
tent, and caspase-3 activation (Fig. 9). Collectively,
these findings suggest that PPARγ activation holds
significant therapeutic potential in prevention and/or
treatment of liver disorders.
Abbreviations
CAT catalase
GPx glutathione peroxidase
HO-1 heme oxygenase-1
Keap1 Kelch-like ECH-associated protein 1
MDA malonaldehyde
Nrf2 nuclear factor erythroid 2-related
factor 2
NQO1 NAD(P)H:quinone oxidoreductase 1
PPARγ peroxisome proliferator-activated
receptor gamma
PPRE PPAR response element
ROS reactive oxygen species
SOD superoxide dismutase
Contributions
L.Z.W., F.C., Y.G.L., and X.F.W. developed the study con-
cept; L.Z.W., F.C., K.L.Z., and Y.Q.A analyzed the data;
F.C., Y.G.L., and X.F.W. acquired the funding; L.Z.W.
wrote the article draft; all authors reviewed and ed-
ited the manuscript.
Funding
This study was supported by Jiaxing Key Discipline
of Medicine – Clinical Pharmacy (2023-ZC-008), Pub-
lic Technology Application Research Program of
Zhejiang Province of China (LGD21H310003), Sci-
ence and Technology Project of Jiaxing of China
(2021AD10023), Natural Science Foundation of Fujian
Province (2024J011357), Natural Science Foundation
of Xiamen (3502Z20227094), Project of Science and
Technology of Xiamen City (3502Z20224ZD1024),
and Xiamen Key Laboratory for Clinical Efficacy
and Evidence-Based Research of Traditional Chinese
Medicine.
ROSIGLITAZONE PROTECTS LIVER CELLS 401
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
Ethics approval and consent to participate
This work does not contain any studies involving hu-
man or animal subjects.
Conflict of interest
The authors of this work declare that they have no
conflicts of interest.
Open access
This article is licensed under a Creative Commons
Attribution 4.0 International License, which permits
use, sharing, adaptation, distribution, and reproduc-
tion in any medium or format, as long as you give
appropriate credit to the original author(s) and the
source, provide a link to the Creative Commons li-
cense, and indicate if changes were made. The im-
ages or other third party material in this article are
included in the article’s Creative Commons license,
unless indicated otherwise in a credit line to the
material. If material is not included in the article’s
Creative Commons license and your intended use
is not permitted by statutory regulation or exceeds
the permitted use, you will need to obtain permis-
sion directly from the copyright holder. To view a
copy of this license, visit http://creativecommons.org/
licenses/by/4.0/.
REFERENCES
1. Yan, Y., Jiang, W., Tan, Y., Zou, S., Zhang, H., Mao, F., Gong, A., Qian, H., and Xu, W. (2017) hucMSC exosome-
derived GPX1 is required for the recovery of hepatic oxidant injury, Mol. Ther., 25, 465-479, https://doi.org/10.1016/
j.ymthe.2016.11.019.
2. Basu, C., and Sur, R. (2018) S-allyl cysteine alleviates hydrogen peroxide induced oxidative injury and apopto-
sis through upregulation of Akt/Nrf-2/HO-1 signaling pathway in HepG2 cells, Biomed. Res. Int., 2018, 3169431,
https://doi.org/10.1155/2018/3169431.
3. Heidari, R., Niknahad, H., Sadeghi, A., Mohammadi, H., Ghanbarinejad, V., Ommati, M. M., Hosseini, A.,
Azarpira, N., Khodaei, F., Farshad, O., Rashidi, E., Siavashpour, A., Najibi, A., Ahmadi, A., and Jamshidzadeh, A.
(2018) Betaine treatment protects liver through regulating mitochondrial function and counteracting oxida-
tive stress in acute and chronic animal models of hepatic injury, Biomed. Pharmacother., 103, 75-86, https://
doi.org/10.1016/j.biopha.2018.04.010.
4. Cichoż-Lach, H., and Michalak, A. (2014) Oxidative stress as a crucial factor in liver diseases, World J. Gastro-
enterol., 20, 8082-8091, https://doi.org/10.3748/wjg.v20.i25.8082.
5. Seen, S. (2021) Chronic liver disease and oxidative stress – a narrative review, Expert. Rev. Gastroenterol.
Hepatol., 15, 1021-1035, https://doi.org/10.1080/17474124.2021.1949289.
6. Chen, Z., Tian, R., She, Z., Cai, J., and Li, H. (2020) Role of oxidative stress in the pathogenesis of nonalcoholic
fatty liver disease, Free Radic Biol. Med., 152, 116-141, https://doi.org/10.1016/j.freeradbiomed.2020.02.025.
7. Xu, P., Cai, X., Zhang, W., Li, Y., Qiu, P., Lu, D., and He, X. (2016) Flavonoids of Rosa roxburghii Tratt ex-
hibit radioprotection and anti-apoptosis properties via the Bcl-2(Ca
2+
)/Caspase-3/PARP-1 pathway, Apoptosis, 21,
1125-1143, https://doi.org/10.1007/s10495-016-1270-1.
8. Esteban-Fernández de Ávila, B., Ramírez-Herrera, D. E., Campuzano, S., Angsantikul, P., Zhang, L., and Wang, J.
(2017) Nanomotor-enabled pH-responsive intracellular delivery of caspase-3: toward rapid cell apoptosis, ACS
Nano, 11, 5367-5374, https://doi.org/10.1021/acsnano.7b01926.
9. Alnahdi, A., John, A., and Raza, H. (2019) Augmentation of glucotoxicity, oxidative stress, apoptosis and mito-
chondrial dysfunction in HepG2 cells by palmitic acid, Nutrients, 11, 1979, https://doi.org/10.3390/nu11091979.
10. Lebovitz, H. E. (2019) Thiazolidinediones: the forgotten diabetes medications, Curr. Diab. Rep., 19, 151, https://
doi.org/10.1007/s11892-019-1270-y.
11. Yen, F. S., Wei, J.C., Chiu, L.T., Hsu, C.C., Hou, M.C., and Hwu, C.M. (2021) Thiazolidinediones were associated
with higher risk of cardiovascular events in patients with type 2 diabetes and cirrhosis, Liver Int., 41, 110-122,
https://doi.org/10.1111/liv.14714.
12. Wallach, J. D., Wang, K., Zhang, A. D., Cheng, D., Grossetta Nardini, H. K., Lin, H., Bracken, M. B., Desai, M.,
Krumholz, H. M., and Ross, J. S. (2020) Updating insights into rosiglitazone and cardiovascular risk through
shared data: individual patient and summary level meta-analyses, BMJ, 368, l7078, https://doi.org/10.1136/
bmj.l7078.
13. Meng, Q., Chen, Z., Gao, Q., Hu, L., Li, Q., Li, S., Cui, L., Feng, Z., Zhang, X., Cui, S., and Zhang, H. (2022)
Rosiglitazone ameliorates spinal cord injury via inhibiting mitophagy and inflammation of neural stem cells,
Oxid. Med. Cell Longev., 2022, 5583512, https://doi.org/10.1155/2022/5583512.
14. Huang, Y., Wang, C., Tian, X., Mao, Y., Hou, B., Sun, Y., Gu, X., and Ma, Z. (2020) Pioglitazone attenuates exper-
imental colitis-associated hyperalgesia through improving the intestinal barrier dysfunction, Inflammation, 43,
568-578, https://doi.org/10.1007/s10753-019-01138-3.
WU et al.402
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
15. Scheen, A. J. (2001) Hepatotoxicity with thiazolidinediones: is it a class effect? Drug Safety, 24, 873-888, https://
doi.org/10.2165/00002018-200124120-00002.
16. Isley, W. L. (2003) Hepatotoxicity of thiazolidinediones, Expert Opin. Drug Safety, 2, 581-586, https://
doi.org/10.1517/14740338.2.6.581.
17. Wang, J. X., Zhang, C., Fu, L., Zhang, D. G., Wang, B. W., Zhang, Z. H., Chen, Y. H., Lu, Y., Chen, X., and Xu, D. X.
(2017) Protective effect of rosiglitazone against acetaminophen-induced acute liver injury is associated with
down-regulation of hepatic NADPH oxidases, Toxicol. Lett., 265, 38-46, https://doi.org/10.1016/j.toxlet.2016.11.012.
18. Cao, Y., He, W., Li, X., Huang, J., and Wang, J. (2022) Rosiglitazone protects against acetaminophen-induced acute
liver injury by inhibiting multiple endoplasmic reticulum stress pathways, BioMed Res. Int., 2022, 6098592,
https://doi.org/10.1155/2022/6098592.
19. Wu, D., Liu, Z., Wang, Y., Zhang, Q., Li, J., Zhong, P., Xie, Z., Ji, A., and Li, Y. (2021) Epigallocatechin-3-gal-
late alleviates high-fat diet-induced nonalcoholic fatty liver disease via inhibition of apoptosis and promo-
tion of autophagy through the ROS/MAPK signaling pathway, Oxid. Med. Cell Longev., 2021, 5599997, https://
doi.org/10.1155/2021/5599997.
20. Shu, M., Lei, W., Su, S., Wen, Y., Luo, F., Zhao, L., Chen, L., Lu, C., Zhou, Z., and Li, Z. (2021) Chlamydia tra-
chomatis Pgp3 protein regulates oxidative stress via activation of the Nrf2/NQO1 signal pathway, Life Sci., 277,
119502, https://doi.org/10.1016/j.lfs.2021.119502.
21. Zhang, H., Davies, K. J. A., and Forman, H. J. (2015) Oxidative stress response and Nrf2 signaling in aging, Free
Radic. Biol. Med., 88, 314-336, https://doi.org/10.1016/j.freeradbiomed.2015.05.036.
22. Deng, S., Liu, S., Jin, P., Feng, S., Tian, M., Wei, P., Zhu, H., Tan,J., Zhao, F., and Gong, Y. (2021) Albumin reduces
oxidative stress and neuronal apoptosis via the ERK/Nrf2/HO-1 pathway after intracerebral hemorrhage in rats,
Oxid. Med. Cell Longev., 2021, 8891373, https://doi.org/10.1155/2021/8891373.
23. Mondal, N. K., Saha, H., Mukherjee, B., Tyagi, N., and Ray, M. R. (2018) Inflammation, oxidative stress, and
higher expression levels of Nrf2 and NQO1 proteins in the airways of women chronically exposed to biomass
fuel smoke, Mol. Cell Biochem., 447, 63-76, https://doi.org/10.1007/s11010-018-3293-0.
24. Ding, X., Jian, T., Li, J., Lv, H., Tong, B., Li, J., Meng, X., Ren, B., and Chen, J. (2020) Chicoric acid ameliorates
nonalcoholic fatty liver disease via the AMPK/Nrf2/NFκB signaling pathway and restores gut microbiota in high-
fat-diet-fed mice, Oxid. Med. Cell Longev., 2020, 9734560, https://doi.org/10.1155/2020/9734560.
25. Roghani, M., Kalantari, H., Khodayar, M. J., Khorsandi, L., Kalantar, M., Goudarzi, M., and Kalantar, H. (2020)
Alleviation of liver dysfunction, oxidative stress and inflammation underlies the protective effect of ferulic acid
in methotrexate-induced hepatotoxicity, Drug Des. Dev. Ther., 14, 1933-1941, https://doi.org/10.2147/dddt.S237107.
26. Yang, J., Fernández-Galilea, M., Martínez-Fernández, L., González-Muniesa, P., Pérez-Chávez, A., Martínez, J. A.,
and Moreno-Aliaga, M. J. (2019) Oxidative stress and Non-alcoholic fatty liver disease: effects of omega-3 fatty
acid supplementation, Nutrients, 11, 872, https://doi.org/10.3390/nu11040872.
27. Chen, Z., Wang, C., Yu, N., Si, L., Zhu, L., Zeng, A., Liu, Z., and Wang, X. (2019) INF2 regulates oxidative stress-
induced apoptosis in epidermal HaCaT cells by modulating the HIF1 signaling pathway, Biomed. Pharmacother.,
111, 151-161, https://doi.org/10.1016/j.biopha.2018.12.046.
28. Fu, J. Y., Jing, Y., Xiao, Y. P., Wang, X. H., Guo, Y. W., and Zhu, Y. J. (2021) Astaxanthin inhibiting oxidative
stress damage of placental trophoblast cells in vitro, Syst. Biol. Reprod. Med., 67, 79-88, https://doi.org/10.1080/
19396368.2020.1824031.
29. Tai,L., Huang, C.J., Choo, K. B., Cheong, S.K., and Kamarul,T. (2020) Oxidative stress down-regulates MiR-20b-5p,
MiR-106a-5p and E2F1 expression to suppress the G1/S transition of the cell cycle in multipotent stromal cells,
Int. J. Med. Sci., 17, 457-470, https://doi.org/10.7150/ijms.38832.
30. Wang, P., Li, B., Cai, G., Huang, M., Jiang, L., Pu, J., Li, L., Wu, Q., Zuo, L., Wang, Q., and Zhou, P. (2014) Acti-
vation of PPAR-γ by pioglitazone attenuates oxidative stress in aging rat cerebral arteries through upregulating
UCP2, J. Cardiovasc. Pharmacol., 64, 497-506, https://doi.org/10.1097/fjc.0000000000000143.
31. Wu, Z., Sainz, A. G., and Shadel, G. S. (2021) Mitochondrial DNA: cellular genotoxic stress sentinel, Trends Bio-
chem. Sci., 46, 812-821, https://doi.org/10.1016/j.tibs.2021.05.004.
32. Nguyen, N. T., Nguyen, T. T., Da Ly, D., Xia, J. B., Qi, X. F., Lee, I. K., Cha, S. K., and Park, K. S. (2020) Oxida-
tive stress by Ca
2+
overload is critical for phosphate-induced vascular calcification, Am. J. Physiol. Heart Circ.
Physiol., 319, H1302-H1312, https://doi.org/10.1152/ajpheart.00305.2020.
33. Li, D., Ni,S., Miao, K. S., and Zhuang,C. (2019) PI3K/Akt and caspase pathways mediate oxidative stress-induced
chondrocyte apoptosis, Cell Stress Chaperones, 24, 195-202, https://doi.org/10.1007/s12192-018-0956-4.
34. Song, J., Lu, C., Zhao, W., and Shao, X. (2019) Melatonin attenuates TNF-α-mediated hepatocytes damage via
inhibiting mitochondrial stress and activating the Akt-Sirt3 signaling pathway, J. Cell Physiol., 234, 20969-20979,
https://doi.org/10.1002/jcp.28701.
ROSIGLITAZONE PROTECTS LIVER CELLS 403
BIOCHEMISTRY (Moscow) Vol. 91 No. 2 2026
35. Ishida,K., Kaji,K., Sato,S., Ogawa,H., Takagi,H., Takaya,H., Kawaratani,H., Moriya,K., Namisaki,T., Akahane,T.,
and Yoshiji, H. (2021) Sulforaphane ameliorates ethanol plus carbon tetrachloride-induced liver fibrosis in mice
through the Nrf2-mediated antioxidant response and acetaldehyde metabolization with inhibition of the LPS/
TLR4 signaling pathway, J. Nutr. Biochem., 89, 108573, https://doi.org/10.1016/j.jnutbio.2020.108573.
36. Cho, R. L., Yang, C. C., Tseng, H. C., Hsiao, L. D., Lin, C. C., and Yang, C. M. (2018) Haem oxygenase-1 up-regu-
lation by rosiglitazone via ROS-dependent Nrf2-antioxidant response elements axis or PPARγ attenuates LPS-me-
diated lung inflammation, Br. J. Pharmacol., 175, 3928-3946, https://doi.org/10.1111/bph.14465.
37. Chiang, M. C., Nicol, C. J., Cheng, Y. C., Lin, K. H., Yen, C. H., and Lin, C. H. (2016) Rosiglitazone activa-
tion of PPARγ-dependent pathways is neuroprotective in human neural stem cells against amyloid-beta-in-
duced mitochondrial dysfunction and oxidative stress, Neurobiol. Aging, 40, 181-190, https://doi.org/10.1016/
j.neurobiolaging.2016.01.132.
38. Kvandová, M., Majzúnová, M., and Dovinová, I. (2016) The role of PPARgamma in cardiovascular diseases,
Physiol. Res., 65, S343-S363, https://doi.org/10.33549/physiolres.933439.
39. Xiong, L., Huang, J., Wang, S., Yuan, Q., Yang, D., Zheng, Z., Wu, Y., Wu, C., Gao, Y., Zou, L., and Hu, G. (2022)
Yttrium chloride-induced cytotoxicity and DNA damage response via ROS generation and inhibition of Nrf2/
PPARγ pathways in H9c2 cardiomyocytes, Arch. Toxicol., 96, 767-781, https://doi.org/10.1007/s00204-022-03225-1.
40. Mahmoud, A. M., Hussein, O. E., Abd El-Twab, S. M., and Hozayen, W. G. (2019) Ferulic acid protects against
methotrexate nephrotoxicity via activation of Nrf2/ARE/HO-1 signaling and PPARγ, and suppression of NF-κB/
NLRP3 inflammasome axis, Food Funct., 10, 4593-4607, https://doi.org/10.1039/c9fo00114j.
41. Li, L., Fu, J., Liu, D., Sun, J., Hou, Y., Chen, C., Shao, J., Wang, L., Wang, X., Zhao, R., Wang, H., Andersen, M. E.,
Zhang, Q., Xu, Y., and Pi, J. (2020) Hepatocyte-specific Nrf2 deficiency mitigates high-fat diet-induced he-
patic steatosis: involvement of reduced PPARγ expression, Redox Biol., 30, 101412, https://doi.org/10.1016/
j.redox.2019.101412.
Publisher’s Note. Pleiades Publishing remains neutral with regard to jurisdictional claims in published
maps and institutional affiliations. AI tools may have been used in the translation or editing of this article.
