BMC Molecular andCell Biology(2024) 25:16
Li et al. BMC Molecular and Cell Biology
https://doi.org/10.1186/s12860-024-00506-8Open Access

RESEARCH

Upregulated dual oxidase 1-induced
oxidative stress and caspase-1-dependent
pyroptosis reflect the etiologies of heart failure
Yan Song Li1,2*, Jingwen Xia1, Chang Yuan Chen1, Shu Hong Ren1 and Mao Rong He1

Abstract
Background Oxidative stress is implicated in the pathogenesis of heart failure. Dual oxidase 1 (DUOX1) might
be important in heart failure development through its mediating role in oxidative stress. This study was designed
to evaluate the potential role of DUOX1 in heart failure.
Materials and methods AC16 cells were treated with 2 µmol/L of doxorubicin (DOX) for 12, 24, and 48 h to construct a heart failure model. DUOX1 overexpression and silencing in AC16 cell were established. DUOX1 expression
was detected by Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Pyroptosis and reactive oxygen species (ROS) production were measured by flow cytometry.
Results Increased DUOX1 expression levels were observed after DOX treatment for 24 h in AC16 cells. DUOX1 silencing inhibited DOX-induced pyroptosis and ROS production. The release of IL-1β, IL-18, and lactate dehydrogenase(LDH), and expression levels of pyroptosis-related proteins were also decreased. DUOX1 overexpression increased
pyroptosis, ROS production, IL-1β, IL-18, and LDH release, and pyroptosis-related protein expression. N-acetyl-cysteine(NAC) significantly reversed DUOX1-induced pyroptosis, ROS, and related factors.
Conclusion These results suggest that DUOX1-derived genotoxicity could promote heart failure development. In
the process, oxidative stress and pyroptosis may be involved in the regulation of DUOX1 in heart failure.
Keywords Reactive oxygen species, Dual oxidase 1, Heart failure, Pyroptosis
Introduction
Heart failure represents the last stage of various diseases
caused by myocardial myofibrillar contractile dysfunction with a syndrome leading to cardiac impairment, andthe disease has been described as an emerging epidemic*Correspondence:
Yan Song Liliyansong1221@163.com
1
Department of Cardiovasology, Shanghai Songjiang District Center
Hospital, NO.748, Zhongshan Middle Road, Songjiang District,
Shanghai 201600, China
2
Department of Cardiovasology, Shanghai Baoshan District Luodian
Hospital, No. 88 Yongshun Road, Baoshan District, Shanghai 201908,
China

for 25 years [1]. Although the survival and pharmacological treatment of the disease have been improved, theprevalence of heart failure remains rising as a result ofa growing and aging population [2, 3]. If a timely treatment is not available, it will pose a serious threat to thepatient’s life and safety. An effective and timely heart failure treatment deserves great attention from clinical medical workers.
It is reported that oxidative stress was one of the maincauses of heart failure development [4]. Oxidative stressis described as a condition of excess reactive oxygen
species (ROS) relative to antioxidant defense and ROS
is known to play a prominent role in the pathogenesisof various cardiac disorders, such as myocardial infarction and heart failure [5]. Study has revealed that ROS© The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which
permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to theoriginal author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images orother third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line
to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory
regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of thislicence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Li et al. BMC Molecular and Cell Biology(2024) 25:16

production is associated with macrophage inflammasome activation, which may directly cause cellular damage, oxidative stress, and pyroptosis, and uncontrolled
ROS production-induced pyroptosis has been observedin various diseases [6–8]. Pyroptosis is a highly regulated
cell death process accompanied by the release of cellular
contents and pro-inflammatory mediators, and is mainly
mediated by various caspases, including caspase-1[9–11]. Caspase-1-dependent pyroptosis, a NOD-likereceptor family pyrin domain containing 3 (NLRP3)mediated classical pathway of pyroptosis, could leadto the activation of NLRP3 inflammasome and downstream caspase-1, IL-1β, and IL-18 [11]. Mature IL-1β is
a pro-inflammatory mediator that mainly modulates theadaptive immune cells by recruiting innate immune cells
to infection sites. Mature IL-18 is significantly related
to interferon-γ production [12]. The functional role ofpyroptosis in cardiovascular diseases (CVDs) has been
widely reported [7, 13, 14].
NAPDH oxidases (NOXs) belong to the family of flavoenzymes, which included a number of isoforms, NOX15, and dual oxidases 1 and 2 (DUOX1-2). NOXs play a
critical role in redox signaling and various CVDs [15].
NADPH oxidases are traditionally known as one of mostimportant proteins associated with ROS generation intissues and cells [16]. For example, in the heart, NOX
2 and 4 are the major sources of ­O2-and ­H2O2, thereby
playing roles in ROS production and consequent cardiac
injury [17, 18]. NOXs are superoxide ­(O2－)-generating
enzymes that catalyze the electron transfer from NADPH
to molecular O­ 2 [16], and the potential role of NOX1 andNOX2 in controlling reperfusion damage were observed[19]. DUOX1 is important in cell signaling, hypoxic
response, and immune function [20]. For example, theupregulation of Duox1 and Duox2 can induce chronic
oxidative stress and fibrosis [21]. Additionally, Duox1
knockdown promotes wound healing by decrease ROS
production [22]. However, the role of DUOX1 in heart
failure is complex and remains unclear.
Various studies focused on the effect of ROS on heart
failure, but only few studied the role of DUOX1. Therefore, in order to evaluate the potential role of DUOX1in heart failure, an in vitro model of heart failure using
doxorubicin (DOX) was constructed. DOX is one of themost effective and broad-spectrum anthracycline antibiotics [23], and it is mainly suitable for acute leukemia,
and is effective for both acute lymphoblastic leukemia
and granulocytic leukemia. It is generally used as a second-line drug, that is, this drug can be considered when
the first-line drug is resistant [24, 25]. As for malignant
lymphoma, it can be used as the drug of choice for alternate use. It also has a certain effect on breast cancer, sarcoma, lung cancer, bladder cancer and other cancers, andPage 2 of 10

they are often used in combination with other anticancer drugs [26]. Additionally, it can induce excessive ROS
production in cardiac mitochondria, leading to acute cardiotoxicity and irreversible degenerative cardiomyopathy[23]. DOX is reported to be widely used in inducing heart
failure in animal studies [27]. DOX-induced cardiotoxicity is characterized by left ventricular dysfunction andcardiac hypertrophy, leading to congestive heart failure.
Here, we attempted to demonstrate the role and mechanism of DUOX1 in regulating DOX-induced heart failure, which may provide an evidence to identify DUOX1
as a potential therapeutic target for heart failure.

Materials and methods
Cell culture

Human cardiomyocyte cell line AC16 was cultured in acell incubator with 5% ­CO2 at 37 °C. Dulbecco’s Modified Eagle’s medium (DMEM, Hyclone, SH30243.01;
Logan, UT, US) was supplemented with 10% fetal bovine
serum (GIBCO, 16000e044; Carlsbad, CA, USA) to culture AC16 cells. Additionally, to avoid bacterial contamination, 1% penicillin with streptomycin (Solarbio,
Beijing, China) was added in the DMEM medium. For
heart failure model, AC16 cells were treated with DOX(Sigma-Aldrich, 25316-40-9) at 2 µmol/L for 12, 24, and48 h. The mRNA level of DUOX1 was quantified using
quantitative real-time polymerase chain reaction (qRTPCR), and the protein levels of DUOX1 were assessed bywestern blot assay.
Construction of vectors with DUOX1 overexpression

The primers for DUOX1 (NM_017434.5) containing
EcoR I and BamH I were designed, and DUOX1 was synthesized based on the designed primer with two kinds ofrestriction enzyme. Then, to construct the DUOX1 overexpression plasmids, the DUOX1 after amplification was
implanted into the vector of pLVX-Puro (Clontech).
After the cells grew to 90% confluency, transfection
was conducted. Based on the manufacturer’s instruction, pLVX-Puro-DUOX1, psPAX2, and pMD2G(Addgen) were co-transfected into 293T cells using Lipofectamine™ 2000 (Invitrogen, CA, USA).
The primers for DUOX1 were designed as follows:
DUOX1-F: 5′-CGG​AAT​TCA​TGG​GCT​TCT​GCC​TGG​
C-3′ (EcoR I).
DUOX1-R: 5′-CGG​GAT​CCC​TAG​AAG​TTC​TCA​TAA​
TGG​TGG​GAG-3′ (BamH I).
Construction and transfection of DUOX1 silencing vectors

A siRNA sequence targeting DUOX1 was synthesized and inserted into the PLKO.1 vector. The lentiviral production system included the expression vectors
psPAX2, pMD2G, and pLKO.1-shDUOX1, which wereLi et al. BMC Molecular and Cell Biology(2024) 25:16

all co-transfected into 293T cells (ATCC) and packaged.
Meanwhile, the plasmid of the scrambled shRNA wasconstructed as the negative control. RNA interference isa powerful tool to induce loss-of-function phenotypes
by inhibiting gene expression at the post-transcriptional
level. One is that dsRNA is cut into siRNA, which allows
a specific mRNA to be degraded; the other is that as longas there is a long dsRNA, it can degrade all RNA andinhibit the synthesis of all proteins. The pLKO.1-puroplasmid inserted the following sequences of DUOX1interference: DUOX1 site 1 (1842–1860): CCA​TCG​TCC​
TTG​AAC​AAT​T; DUOX1 site 2 (3085–3103): GGA​ACT​GAC​ATG​GGA​AGA​T; and DUOX1 site 3 (4572–4590):
GCT​GCC​AAG​TGT​TCT​GTA​A .
qRT‑PCRTo quantify the mRNA levels of DUOX1, the total RNA
in cells was extracted using a TRIzol reagent (Invitrogen,1596-026). After quantifying and confirming the RNA
integrity, the RNA samples were reverse-transcribed tocDNA; then, cDNA was amplified using SYBR GreenqPCR Master Mixes (Thermo Fisher, Rockford, IL, USA).
The Ct value was defined as the number of cycles thatthe fluorescent signal in the reaction tube reaches the set
threshold. The relative DUOX1 mRNA levels normalized
to GAPDH were calculated using the ­2−△△Ct method.
The primer sequences for DUOX1 were designed as follows: Primer F: 5′ AGT​GCC​TGG​ATT​GTT​GCC​ 3′ andPrimer R: 5′ TTG​CTG​GAC​AGG​ACG​AAG​ 3′. The
primer sequences for GAPDH were designed as follows:
Primer F: 5′ GGA​TTG​TCT​GGC​AGT​AGC​C 3′ andPrimer R: 5′ ATT​GTG​AAA​GGC​AGG​GAG​ 3′.
Western blot analysis

The total protein from AC16 cells was extracted usinga RIPA buffer (Solarbio, R0010, Beijing, China), and theprotein level was determined by BCA protein assay kit(Thermo Fisher Scientific, PICPI23223). The sample
was boiled at 95 °C for 10 min and separated throughthe gels with 10% concentration of SDS-PAGE gel(JRDUN Biotechnology Co., Ltd, Shanghai, China).
Then, the separated proteins were transferred to polyvinylidene fluoride membrane and blocked with 5%
nonfat milk. After blocking, the membrane was further
incubated with anti-DUOX1, anti-active caspase-1, antiGAPDH (67226-1-lg, 22915-1-AP, 60004-1-1G, Proteintech), anti-apoptosis-associated speck-like protein
containing a CARD (ASC), anti-NLRP3, anti-pro-caspase-1, and anti-Gasdermin D-N domain (GSDMD-N)(Ab155970, Ab263899, Ab179515, Ab215203, Abcam)
overnight at 4 °C with gentle shaking. After washing
thrice by TBST(TBS + Tween20), the polyvinylidene
fluoride membrane was incubated with horseradish

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peroxidase-conjugated secondary antibodies (1:1000,
Beyotime, Shanghai, China) for 1 h at 25 °C. Finally,
the expression levels of proteins were measured using achemiluminescent imaging system (Tanon 5200, Shanghai, China).
ROS detection

To detect the ROS production, Active Oxygen Detection
Kit (S0033, Beyotime) was employed. First, the cell pellets were obtained after centrifugation and resuspended
using 1 mL of cooled PBS. Following the manufacturer’s
instruction, the DCFH-DA staining working solution was
diluted to 10 µM using a serum-free medium. Then, the
DCFH-DA probe solution was added to the cell suspension in the dark for 20 min at 37 °C. To make the probe
in full contact with the cells, the samples were mixed
every 3–5 min for 20 min. Then, the cells were washedthrice using a serum-free medium to remove the DCFHDA probe that did not enter the cells. Finally, the samples
were detected by flow cytometry (BD Biosciences, Franklin Lake, NJ, USA).
Pyroptosis assay

To detect the level of pyroptosis, the cells were resuspended in PBS. Then, active caspase-1 (1:30; EL900443,
EterLife) was added in the cell incubation for 1 h at 25 °C.
To remove non-combined caspase-1, the cells were
washed with PBS for three times and incubated with propidium iodide solution (PI, P3566; Invitrogen) at 3 µM
for 15 min in the dark. Finally, the rates of cell pyroptosis
were estimated based on flow cytometry.
Enzyme‑linked immunosorbent assay (ELISA)

The secreted IL-1β and IL-18 in the supernatants weredetected using ELISA. The cells were collected by centrifuging at 3000 rpm/min for 20 min. After removing thecell pellets, the secreted IL-1β and IL-18 were quantified
using Human IL-1β and IL-18 ELISA kits (NeoBioscience, Shenzhen, Guangdong, China), following the manufacturer’s instructions. The amount of IL-1β and IL-18
was detected at 450 nm by a microplate reader (Bio-Rad,
Hercules, CA, USA).
Biochemical detection

The lactate dehydrogenase (LDH) in supernatant and cellular protein level was detected using the LDH (A020-1)
Kit and BCA (A045-3) Kit (Nanjing, Jiancheng Biotechnology Research Institute, Jiangsu, China). The supernatant and kit solution were mixed for 15 min at 37 °C ina water bath. Then, the LDH and cellular protein were
measured at 440 nm (LDH) and 562 nm (BCA).

Li et al. BMC Molecular and Cell Biology(2024) 25:16

Statistical analysis

Each experiment was performed in triplicate. The data
were presented as mean value ± standard deviation of
three independent biological replicates. To compare
the difference in mean values among the groups, oneway analysis of variance with Tukey’s post hoc tests wasapplied. P < 0.05 was regarded as statistically significant.
All statistical analyses in the study were performed using
the GraphPad Prism 7.0 software (San Diego, CA, USA).

Results
DUOX1 was highly expressed in DOX‑induced AC16 cells
in a time‑dependent manner

Human cardiac AC16 cells were treated with 2 µmol/Lof DOX to simulate and construct a heart failure modelin vitro. qRT-PCR and western blot detection revealed
that the expression of DUOX1 increased in a timedependent manner (Fig. 1), that is, DUOX1 expression
increased significantly as time increased. Further, 24 h
was chosen as the DOX treatment time.
DUOX1 silencing potently reversed DOX‑induced
pyroptosis and ROS production

The overexpression and interference efficiency of DUOX1
was examined by qRT-PCR and western blot on mRNAand protein levels, respectively. Both mRNA and protein levels of DUOX1 were significantly increased in the
DUOX1 overexpression (oeDUOX1) group (Fig. 2A-C),
and decreased in the group of shDUOX1-1, shDUOX1-2,
shDUOX1-3 as compared with shNC (Fig. 2D-F), suggesting the model of DUOX1 overexpression and silencing were successfully constructed (Fig. 2).
After efficiency appraisal, the shDUOX1-1 andshDUOX1-2 vector was transfected to AC16 cells,
respectively. Then DOX (2 µmol/L) was added into

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the cells. As shown in Fig. 3, DOX treatment led to anincrease ROS production (Fig. 3A) and pyroptosis(Fig. 3B) in AC16 cells. While, silencing DUOX1 significantly reversed the increased ROS production andpyroptosis introduced by DOX (Fig. 3).
DUOX1 silencing potently reversed DOX‑induced
inflammation and abnormal protein expression

Further, we have investigated the effects of DUOX1knockdown on IL-1β and IL-18 secretion, and LDH
release, as well as protein level of pyroptosis associated pro- and active caspase-1, GSDMD-N, and NLRP3.
The results indicated that DOX treatment led to IL-1β(Fig. 4A) and IL-18 secretion (Fig. 4B). The CIK cell killing activity was detected using LDH release test, andLDH release was significantly increased after DOX treatment (Fig. 4C), suggesting the increased CIK cell activity. After DUOX1 silencing, IL-1β and IL-18 secretion,and LDH release were significantly decreased (Fig. 4AC). In addition, the protein expression of pro- and active
caspase-1, GSDMD-N, and NLRP3 was detected using
western blot. Figure 4D showed that the expression levelsof pro- and active caspase-1, GSDMD-N, and NLRP3 in
AC16 cells were all increased after DOX treatment, and
DUOX1 silencing reversed the increased levels of thementioned proteins (Fig. 4D).
DUOX1 promoted the pyroptosis in AC cells probably
through ROS regulation

In subsequent experiments, we have examined the effects
of DUOX1 overexpression on AC16 cells, and the participation of ROS was investigated during this process.
AC16 cells was treated by DUOX1 overexpression andthe ROS scavenger NAC (100 µM) for 24 h, and then
the ROS production, pyroptosis, IL-1β and IL-18 secretion, LDH release, and the protein expression of pro- andFig. 1 DUOX1 was dependently induced by DOX in AC16 cells. Human cardiac AC16 cells were treated with 2 µmol/L of DOX to simulate
and construct an in vitro model of heart failure. qRT-PCR (A) and western blot (B) detected the expression of DUOX1 at different time points.
**p < 0.01. The blots were cut prior to hybridisation with antibodies during immunoblotting experiments

Li et al. BMC Molecular and Cell Biology(2024) 25:16

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Fig. 2 The efficiency identification of DUOX1 overexpression and knockdown. DUOX1 overexpression and interference lentiviral vector
was transfected into myocardial AC16 cells, and the overexpression (A-C) and interference (D-F) efficiency of DUOX1 was detected by qRT-PCR
and western blot. **p < 0.01 vs. vector/shNC. The blots were cut prior to hybridisation with antibodies during immunoblotting experiments

Fig. 3 DUOX1 knockdown reversed DOX-induced ROS production and pyroptosis. A DUOX1 knockdown reversed DOX-induced ROS production,
tested by flow cytometry. B DUOX1 knockdown reversed DOX-induced pyroptosis, tested by flow cytometry. **p < 0.01 vs. control; ^^p < 0.01 vs.
shNC + DOX (2 µmol/L)Li et al. BMC Molecular and Cell Biology(2024) 25:16

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Fig. 4 DUOX1 knockdown reversed DOX-induced inflammation and abnormal protein expression. The secretion of IL-1β (A) and IL- 18 (B) in the cellsupernatant was detected by ELISA. C LDH activity was detected by biochemical method. D Western blot was used to detect the expressionof pyroptosis indexes active caspase-1, pro-caspase-1, NLRP3, and GSDMD-N. **p < 0.01 vs. control; ^^p < 0.01 vs. shNC + DOX (2 µmol/L). The blots
were cut prior to hybridisation with antibodies during immunoblotting experiments

active caspase-1, GSDMD-N, and NLRP3 were tested.
The results showed the ROS production, and pyroptosis
were induced by DUOX1 overexpression (Fig. 5A, B).
Meanwhile, DUOX1 overexpression increased IL-1β and
IL-18 secretion, LDH release, protein levels of pro- andactive caspase-1, GSDMD-N, and NLRP3 (Fig. 6A-D).
Above all, these changes induced by DUOX1 overexpression were all reversed by NAC treatment (Figs. 5 and
6). Suggested that ROS mediated DUOX1 induced cell
pyroptosis.

Discussion
Up to now, heart failure remains a major cause of mortality, morbidity, and poor quality of life. To maintainthe normal function of cardiovascular system, a balance
between cell formation and death is needed [28]. DUOX,
as a member of NADPH oxidase family, participateddirectly in generating hydrogen peroxide/ROS [29]. In
normal condition, there is a balance between ROS andantioxidants, but the excessive amounts of ROS would
result in various CVDs, including heart failure [4]. In
this study, we found the increased DUOX1 expression
levels in DOX-induced cellular heart failure model, suggesting that DUOX1 may contribute to heart failure
development.
The underlying mechanism of DUOX1 in regulatingheart failure has been also explored. It is well-known
that heart failure development is associated with oxidative stress, which might be caused by the complex effects
of ROS [4]. DUOX1-induced ROS production has also
been shown in various diseases, such as DUOX1 nasal
polyposis, allergy and asthma, chronic obstructive pulmonary disease, lung fibrosis and cancer [30]. Consistent with these reports, we found that the increased ROS

Li et al. BMC Molecular and Cell Biology(2024) 25:16

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Fig. 5 DUOX1 promoted ROS production and pyroptosis were inhibited by NAC. ROS scavenger NAC treatment reversed DUOX1 overexpression
induced ROS production (A) and pyroptosis (B). Flow cytometry was designed to detect cell ROS level and pyroptosis. **p < 0.01 vs. vector;^^p < 0.01 vs. oeDUOX1 + NAC (100 µmol/L)

production were associated with DUOX1 expression.
Silencing DUOX1 potently reversed DOX-induced ROS
production. Meanwhile, NAC, a kind of active oxygen inhibitor, significantly reversed the level of ROS,
and related factors induced by DUOX1 overexpression,suggesting a direct oxidative stress effect of increased
DUOX1 expression on heart failure.
Furthermore, the relationship between pyroptosis andpathogenesis of various CVDs, including heart failure,has been demonstrated [31], and the inhibition of pyroptosis was recognized as a potential therapeutic intervention to prevent CVDs [32]. Consistent with these, ourdata showed that the increased pyroptosis was associated with DUOX1 expression. Silencing DUOX1 potently
reversed DOX-induced pyroptosis. From these, we speculated that pyroptosis may be involved in DUOX1 regulation in DOX-induced heart failure. The Studies also
reported that elevated mRNA of NLRP3 and enhanced
cleavage of caspase-1 were observed along with cardiac
hypertrophy and ventricular dilatation in calcineurintransgene (CNTg) mice, an established mouse model forchronic heart failure [33]. Genetic ablation of NLRP3 oradministration of IL-1 receptor antagonist can reduce
cardiac inflammation and systolic dysfunction [34–36].

In line with these previous reports, the data in this study
showed the increased caspase-1 expression when thelevel of pyroptosis was high. Caspase-1 is one of the proinflammatory family of caspases, is a necessary ingredient in the activation of pro-IL-1β and pro-IL-18 [37]. In
the pathophysiology of heart failure, previous data demonstrated the critical roles of active IL-1β and IL-18 [38,39]. This study showed the increased secretion of IL-1β
and IL-18 when DUOX1 was overexpressed in AC16
cells, suggesting that the caspase-1-dependent pyroptosis in heart failure was triggered by the upregulation of
DUOX1. Meanwhile, NAC significantly reversed the levels of pyroptosis, and related factors induced by DUOX1
overexpression, suggesting that DUOX1 regulates caspase-1-dependent pyroptosis in AC16 cells may throughmodulating ROS.
NLRP3-mediated pyroptosis pathway are widely studied, and related inhibitors are well used in clinical practice in cardiovascular disease. Our results illustrate thatsuppressing ROS production by DUOX1 knockdown wasan effective strategy to decrease NLRP3-mediated myocardial cells injury. These suggest that similar function of
DUOX1 are very likely reproduced in other cardiovascular disease, such as myocardial ischemia-reperfusion
injury and myocardial infarction. Meanwhile, the role ofLi et al. BMC Molecular and Cell Biology(2024) 25:16

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Fig. 6 DUOX1 promoted the pyroptosis of AC16 cells probably through ROS regulation. The secretion of IL-1β (A) and IL-18 (B) in the cellsupernatant was detected by ELISA. C LDH activity was detected by biochemical method. D Western blot was used to detect the expressionof active caspase-1, pro-caspase-1, NLRP3, and GSDMD-N. **p < 0.01 vs. vector; ^^p < 0.01 vs. oeDUOX1 + NAC (100 µmol/L). The blots were cut prior
to hybridisation with antibodies during immunoblotting experiments

DUOX1 in myocardial injury and ventricular remodelingneeds more practice and exploration.

pathways associated with DUOX1 underlying heart
failure.

Conclusions
Our study revealed the important role of DUOX1 inheart failure. DUOX1 inhibition in heart failure might
prevent the damage induced by oxidative stress andpyroptosis, which might be a novel therapeutic biomarker for heart failure. However, our conclusion was
based on the experiment in vitro. Future studies on
animal model and primary myocardial cells should bedesigned to provide more insight into the biological

Supplementary Information
The online version contains supplementary material available at https://​doi.​
org/​10.​1186/​s12860-​024-​00506-8.
Supplementary Material 1.
Acknowledgements
Not applicable.
Authors’ contributions
YSL conceived and designed the study. JWX, CYC, SHR and MRH performed
the experiments, and collected and analyzed the data. YSL wrote theLi et al. BMC Molecular and Cell Biology(2024) 25:16

manuscript. All authors read and approved the final manuscript and agree tobe accountable for all aspects of the research in ensuring that the accuracy or
integrity of any part of the work are appropriately investigated and resolved.
Funding
This study was supported by Songjiang District New round of Medical key
discipline Construction Project (ZK2019A07).
Availability of data and materials
The data are available from the corresponding author upon reasonable
request.

Declarations
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare no conflict of interest.
Received: 8 May 2023 Accepted: 8 March 2024

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