International Immunopharmacology 64 (2018) 238–245

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International Immunopharmacology
journal homepage: www.elsevier.com/locate/intimp

Isofraxidin inhibits interleukin-1β induced inflammatory response in human T

osteoarthritis chondrocytes
Jian Lin1, Xiaobin Li1, Weihui Qi, Yingzhao Yan, Kai Chen, Xinghe Xue, Xinxian Xu,
⁎
Zhenhua Feng, Xiaoyun Pan
Department of Orthopaedic, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325000, China
Key Laboratory of Orthopaedics of Zhejiang Province, Wenzhou 325000, China
The Second School of Medicine, Wenzhou Medical University, China

A R T I C LE I N FO A B S T R A C T

Keywords: Osteoarthritis (OA) is the most prevalent disease of knee especially in the aged people. Isofraxidin (IF) is a
Osteoarthritis coumarin compound refined from traditional Chinese medicines with potential anti-inflammatory ability. This
Inflammation study aimed to evaluate protective anti-inflammatory effects of IF in human OA chondrocytes. The chondrocytes
Isofraxidin were isolated from OA patients and pretreated with IF before treatment with IL-1β. The results showed that IF
Chondrocytes
blocked IL-1β-stimulated production of NO and PGE2. In addition, IF inhibited the expression of COX-2, iNOs,
NF-κB
MMP-1, MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5, and increased the levels of aggrecan and collagen-II.
Mechanistically, IF suppressed IL-1β-induced IκB-α degradation and NF-κB activation. In conclusion, our results
demonstrate that IF inhibits inflammation in OA via the regulation of NF-κB signaling, and suggest that IF may
be a potential therapeutic agent for OA.

1. Introduction
Osteoarthritis (OA) is a common deteriorating joint pathologic
process caused by joint instability and it is estimated that 12.1% of U.S.
population aged 25 and older have OA. OA could cause serious joint
pain and irreversible loss of joint movement, and OA is characterized by
erosion of the articular cartilage, synovium inflammation, subchondral
bone sclerosis and osteophyte formation [1,2]. Recent studies showed
that inflammation and inflammatory factors play important role in the
progression of OA [3,4]. Interleukin-1β (IL-1β), the major proin-
flammatory cytokine, could be involved in the destruction of cartilage
[5,6]. IL-1β could induce the production of matrix metalloproteinases
(MMPs) and a disintegrin and metalloproteinase with thrombospondin
motifs (ADAMTS), which are responsible for the downregulation of type
II collagen and proteoglycans in articular cartilage [7,8]. Furthermore,
treatment of chondrocytes with IL-1β could stimulate the release of
inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(COX-2),
leading to the production of nitric oxide (NO) and prostaglandin E2
(PGE2) [9,10]. Therefore, these inflammatory mediators may be ef-
fective therapeutic targets for OA.
Isofraxidin (IF, 7-hydroxy-6, 8-dimethoxycoumarin) is a coumarin
compound isolated from traditional Chinese herbs usually used for anti-
tumor, anti-oxidant, anti-bacterial, and anti-inflammatory treatments
[11–14]. IF has multifarious biological activities, including anti-oxi-
dant, anti-bacterial and anti-inflammatory actions [15–17]. IF reduced
the levels of iNOS, COX-2, and tumor necrosis factor-α (TNF-α) by
regulating inflammatory modulators, such as inhibitory kappa B (IκBα)
and nuclear factor-kappa B (NF-κB) [18]. IF has been reported to ab-
rogate LPS-induced oxidative stress and inflammation by inhibiting
TNF-α production and MAPK pathway in mouse peritoneal macro-
phages [19]. In addition, IF exerted anti-inflammatory effect via NF-κB
suppression in vivo [20]. However, anti-inflammatory effect of IF in OA
remains unknown. Therefore, in this study we evaluated the anti-in-
flammatory effects and explored the possible mechanism of IF on IL-1β-
induced inflammation in human OA chondrocytes in vitro.
2. Materials and methods
2.1. Reagents and antibodies
Isofraxidin (purity > 98%), collagenase type II, recombinant human
IL-1β, and dimethylsulfoxide (DMSO) were purchased from Sigma

⁎
Corresponding author at: Department of Orthopaedic, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou
325000, China.
E-mail address: panxy9046@163.com (X. Pan).
1
Jian Lin and Xiaobin Li contribute equally to this work.

https://doi.org/10.1016/j.intimp.2018.09.003
Received 4 June 2018; Received in revised form 1 September 2018; Accepted 5 September 2018
1567-5769/ © 2018 Elsevier B.V. All rights reserved.
J. Lin et al. International Immunopharmacology 64 (2018) 238–245

Table 1
Primers used in this study.
Gene Forward primer Reverse primer

COX-2 5′-GAGAGATGTATCCTCCCACAGTCA-3′ 5′-GACCAGGCACCAGACCAAAG-3′

iNOS 5′-CCTTACGAGGCGAAGAAGGACAG-3′ 5′-CAGTTTGAGAGAGGAGGCTCCG-3′
MMP-1 5′-GGGAATAAGTACTGGGCTGTTCAG-3′ 5′-CCTCAGAAAGAGCAGCATCGATATG-3′
MMP-3 5′-CTGGCCTGCTGGCTCATGCTT-3′ 5′-GCAGGGTCCTTGGAGTGGTCA-3′
MMP-13 5′-CCAGAACTTCCCAACCAT-3′ 5′-ACCCTCCATAATGTCATACC-3′
ADAMTS-4 5′-CCTGGCAAGGACTATGATGCTGA-3′ 5′-GGGCGAGTGTTTGGTCTGG-3′
AMAMTS-5 5′-GCAGAACATCGACCAACTCTACTC-3′ 5′-CCAGCAATGCCCACCGAAC-3′
Collagen-II 5′-CTCAAGTCGCTGAACAACCA-3′ 5′-GTCTCCGCTCTTCCACTCTG-3′
Aggrecan 5′-AAGTGCTATGCTGGCTGGTT-3′ 5′-GGTCTGGTTGGGGTAGAGGT-3′
GAPDH 5′-TCTCCTCTGACTTCAACAGCGAC-3′ 5′-CCCTGTTGCTGTAGCCAAATTC-3′

Fig. 1. Effect of IF on human OA chondrocyte viability. The cells were treated

with IF (0, 1, 10, 50, 100 μM) for 24 h. The cell viability was determined by
CCK-8 assay. The values are mean ± SD of three independent experiments.
*p < 0.05 compared with control group.

Chemical Co. (St. Louis, MO, USA). Primary antibodies against iNOs,
COX-2, and collagen-II were acquired from Abcam (Cambridge, MA,
USA). Primary antibodies against p65, p-p65, IkBα, and p-IkBα were
purchased from Cell Signaling Technology (Beverly, MA, USA). Cell-
Counting Kit-8 (CCK-8) was obtained from Dojindo (Kumamo, Japan).
TRIzol reagent was purchased from Invitrogen (Carlsbad, CA, USA).
QuantiTect Reverse Transcription Kit was purchased from Qiagen
(Valencia, CA, USA). Fetal bovine serum (FBS), bovine serum albumin
(BSA), and Dulbecco's modified Eagle's medium (DMEM)/F12 medium
were purchased from Gibco (Life Technologies Corp., Carlsbad, CA,
USA). SYBR Green Master Mix was purchased from Bio-Rad
Laboratories (CA, USA). ELISA kits of PGE2, MMP-1, MMP-3, MMP-13,
ADAMTS-4, and ADAMTS-5 were purchased from R&D Systems
(Minneapolis, MN, USA). Griess reagent was purchased from Beyotime
Institute of Biotechnology (Shanghai, China).

Fig. 2. Effect of IF on IL-1β-induced NO and PGE2 production in human OA

2.2. Primary human chondrocyte culture
Tissue collection was according to the terms of the Medical Ethical
Committee of the Second Affiliated Hospital, Wenzhou Medical
University and following the guidelines of the Declaration of Helsinki.
Human cartilage samples were obtained from four OA patients (aged
56–66 years, two men and two women) who were diagnosed as os-
teoarthritis [21]. All patients underwent total knee replacement surgery
at the Second Affiliated Hospital of Wenzhou Medical University, and
provided ethical consent. Cartilage tissues were isolated from bone and
connective tissues, cut into small pieces, washed three times with PBS,
digested with 0.25% trypsin–EDTA solution, and then incubated with
2 mg/ml (0.1%) collagenase II for 4 h at 37 °C. After centrifuged at
1000 rpm for 5 min, the digested cartilage tissues were suspended and
seeded into tissue culture flasks. The chondrocytes were cultured in
DMEM/F12 with 10% FBS and 1% antibiotic mixture in the incubator
with 5% CO2 at 37 °C. The medium was changed every two days, and
the cells were passaged by using 0.25% Trypsin-EDTA solution. Only
chondrocytes. Chondrocytes were pretreated with various concentrations of IF
(1, 10, 50 μM) and then treated with or without IL-1β (10 ng/ml) for 24 h. NO
levels were determined by Griess reaction (a), and the levels of PGE2 were
assessed by ELISA (b). The values are mean ± SD of three independent ex-
periments. #p < 0.05 compared with control group. *p < 0.05, **p < 0.01
compared with IL-1β group.
passage 1 to 2 was used in our study to avoid the phenotype loss.
2.3. Cell viability
Cell viability was tested using CCK-8 kit according to the manu-
facturer's instructions. Briefly, human OA chondrocytes were seeded in
96-well plates (6000 cells/well) and treated with or without IF (1, 10,
50, 100 μM) for 24 h. After that, 10 uL CCK-8 was added to each well
and incubated at 37 °C for 4 h. The absorbance of wells was then
measure using a microplate reader (Leica Microsystems, Germany).

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Fig. 3. Effect of IF on IL-1β-induced iNOS and COX-2 expression in human OA chondrocytes. Human OA chondrocytes were pretreated for 2 h with various
concentrations of IF (1, 10, 50 μM) and then stimulated or not stimulated with IL-1β (10 ng/ml) for 24 h. The mRNA expression levels of COX-2 (a) and iNOS (b) were
assessed by qRT-PCR. The protein expression levels of iNOS and COX-2 were determined by Western blot analysis (c–e). The values are mean ± SD of three
independent experiments. #p < 0.05 compared with control group. *p < 0.05, **p < 0.01 compared with IL-1β group.

Fig. 4. Effect of IF on IL-1β-induced MMP-1, MMP-3 and MMP-13 expression in human OA chondrocytes. Human OA chondrocytes were pretreated for 2 h with
various concentrations of IF (1, 10, 50 μM) and then stimulated or not stimulated with IL-1β (10 ng/ml) for 24 h. The mRNA expression levels of MMP-1 (a), MMP-3
(b), and MMP-13 (c) were assayed by qRT-PCR. The protein levels of MMP-1, MMP-3, and MMP-13 in culture medium were determined by ELISA (d-f). The values are
mean ± SD of three independent experiments. #p < 0.05 compared with control group. *p < 0.05, **p < 0.01 compared with IL-1β group.

2.4. Griess reaction and ELISAs
The content of NO in culture medium was measured by Griess re-
agent as previously described [22]. The levels of PGE2, MMP-1, MMP-3,
MMP-13, ADAMTS-4, and ADAMTS-5 in culture medium were eval-
uated using commercial ELISA kit according to the manufacturer's in-
structions.
2.5. qRT-PCR
Total RNA was extracted from cells using TRIzol reagent. First-
strand cDNA was synthesized using 1 μg of total RNA and the
QuantiTect Reverse Transcription kit. Quantitative real-time PCR
(qPCR) was performed using CFX96 Real-Time PCR System (Bio-Rad
Laboratories, CA, USA) under the following conditions: 10 min 95 °C,
followed by 40 cycles of 15 s 95 °C and 1 min 60 °C. The reaction was
performed in a total volume of 10 μl, containing 4.5 μl diluted cDNA,
0.25 μl forward primer, 0.25 μl reverse primer and 5 μl SYBR Green
Master Mix. The level of target mRNA was normalized to the level of
GAPDH and compared with control. Data were analyzed using the
2−ΔΔCT method [23]. The primers were designed using GenScript Pcr
Primer Design online program and the sequences were listed in Table 1.

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2.6. Western blot analysis
The total protein was extracted from chondrocytes using RIPA lysis
buffer supplemented with 1 mM PMSF (Phenylmethanesulfonyl
fluoride) on the ice for 10 min followed by 15 min centrifugation at
12,000 rpm at 4 °C, and then protein concentration was measured using
the BCA protein assay kit (Beyotime). 40 μg of total protein were re-
solved on 12% SDS-PAGE and transferred to PVDF membranes.
Membranes were blocked with 5% nonfat milk for 2 h at room tem-
perature and immediately washed three times for 5 min in TBS with
Tween-20 (TBST). Next, the membranes were incubated with primary
antibody against COX-2, iNOs, p65, p-p65, IκB-α, p-IκB-α or β-actin
(dilution 1:1000) overnight at 4 °C, and washed three times with TBST
for 5 min. The membranes were incubated subsequently with respective
secondary antibodies for 2 h at room temperature. After washing with
TBST, the blots were visualized by electrochemiluminescence plus re-
agent (Invitrogen). Finally, the intensity of the blots was quantified
with Image Lab 3.0 software (Bio-Rad).
2.7. Immunofluorescence
The chondrocytes were seeded in glass plated in a six-well plate and
incubated for 24 h. Glass coverslips with chondrocyte monolayers were
rinsed three times in PBS before fixation in 4% paraformaldehyde and
permeation in 0.1% Triton X-100 diluted in PBS for 15 min. Next, the
cells were blocked with 5% goat serum albumin for 1 h at 37 °C, rinsed
with PBS and incubated with primary antibody for collagen-II or p65
(1:200) in a humid chamber overnight at 4 °C. On the next day, cells
were incubated with fluorescein-conjugated goat anti-rabbit IgG anti-
body (1:500) for 1 h at 37 °C. Afterwards, cells were washed three times
with PBS and then labeled with DAPI (Invitrogen) for 1 min. Finally,
slides were observed under a fluorescence microscope (Olympus Inc.,
Tokyo, Japan).
2.8. Statistical analysis
All experiments were at least performed triplicately. Data are
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International Immunopharmacology 64 (2018) 238–245
Fig. 5. Effect of IF on IL-1β-induced
ADAMTS-4 and ADAMTS-5 expression in
human OA chondrocytes. Human OA
chondrocytes were pretreated for 2 h with
various concentrations of IF (1, 10, 50 μM)
and then stimulated or not stimulated with
IL-1β (10 ng/ml) for 24 h. The mRNA ex-
pression levels of ADAMTS-4 (a), and
ADAMTS-5 (b) were assayed by qRT-PCR.
The protein levels of ADAMTS-4, and
ADAMTS-5 in culture medium were de-
termined by ELISA (c, d). The values are
mean ± SD of three independent experi-
ments. #p < 0.05 compared with control
group. *p < 0.05, **p < 0.01 compared
with IL-1β group.
expressed as mean ± standard deviation (SD). Statistical analyses were
performed using SPSS statistical software program 20.0. Statistical
significance was analyzed by one-way analysis of variance (ANOVA)
followed by Tukey's post-hoc test if necessary. P < 0.05 was con-
sidered statistically significant.
3. Results
3.1. Effect of IF on human OA chondrocyte viability
The cytotoxic effect of IF on chondrocytes was determined at var-
ious concentrations (1, 10, 50, 100 μM) using CCK-8. As shown in
Fig. 1, 100 μM IF significantly reduced cell viability after treatment for
24 h, but cell viability was not significantly affected by IF at con-
centrations up to 50 μM. Thus we chose IF at concentrations of 1, 10
and 50 μM in subsequent experiments.
3.2. Effect of IF on NO and PGE2 production in human OA chondrocytes
Chondrocytes were pretreated with various concentration of IF (1,
10, 50 μM) for 2 h, then treated with IL-1β (10 ng/ml) for 24 h. As
shown in Fig. 2, the levels of NO and PGE2 in cell culture medium
increased significantly in IL-1β treated group compared to control
group, but IF significantly reduced IL-1β-stimulated production of NO
and PGE2 in a concentration-dependent manner. IF alone had no sig-
nificant effects on NO and PGE2 production in the chondrocytes
(Supplemental Fig. 1).
3.3. Effect of IF on iNOS and COX-2 expression in human OA chondrocytes
Next we detected the expression of iNOS and COX-2 in chon-
drocytes. We found that IL-1β stimulation led to the upregulation of
iNOS and COX-2 mRNA expression compared with the control group.
However, IF decreased the expression of iNOS and COX-2 at mRNA
level dose-dependently (Fig. 3a, b). Similarly, protein expression levels
of iNOS and COX-2 were significantly stimulated by IL-1β but decreased
by IF pretreatment (Fig. 3c–e). IF alone had no significant effects on
J. Lin et al. International Immunopharmacology 64 (2018) 238–245

Fig. 6. Effect of IF on IL-1β-induced aggrecan and collagen-II expression in human OA chondrocytes. Human OA chondrocytes were pretreated for 2 h with various
concentrations of IF (1, 10, 50 μM) and then stimulated or not stimulated with IL-1β (10 ng/ml) for 24 h. The expression of collagen-II was visualized by im-
munofluorescence (a). The mRNA expression levels of aggrecan (b) and collagen-II (c) were assayed by qRT-PCR. The values are mean ± SD of three independent
experiments. #p < 0.05 compared with control group. *p < 0.05, **p < 0.01 compared with IL-1β group.

iNOS and COX-2 expression in the chondrocytes (Supplemental Fig. 2).
3.4. Effect of IF on MMP-1, MMP-3 and MMP-13 expression in human OA
chondrocytes
As shown in Fig. 4a–c, IL-1β significantly upregulated mRNA ex-
pression of MMP-1, MMP-3 and MMP-13 in chondrocytes compared
with control group. IF remarkably decreased mRNA levels of MMP-1,
MMP-3 MMP-13 in chondrocytes stimulated with IL-1β. ELISA assay
showed that the secretion of MMP-1, MMP-3 and MMP-13 in culture
medium stimulated by IL-1β was significantly inhibited by IF
(Fig. 4d–f). IF alone had no significant effects on MMP-1, MMP-3 and
MMP-13 expression in the chondrocytes (Supplemental Fig. 3).
3.5. Effect of IF on ADAMTS-4 and ADAMTS-5 expression in human OA
chondrocytes
As shown in Fig. 5a, b, IL-1β significantly upregulated mRNA ex-
pression of ADAMTS-4 and ADAMTS-5 in chondrocytes compared with
control group. IF remarkably decreased mRNA levels of ADAMTS-4 and
ADAMTS-5 in chondrocytes stimulated with IL-1β. ELISA assay showed
that the secretion of ADAMTS-4 and ADAMTS-5 in culture medium
stimulated by IL-1β was significantly inhibited by IF (Fig. 5c, d). IF
alone had no significant effects on ADAMTS-4 and ADAMTS-5 expres-
sion in the chondrocytes (Supplemental Fig. 3).
3.6. Effect of IF on aggrecan and collagen-II expression in human OA
chondrocytes
Immunofluorescence analysis showed that IL-1β stimulation led to
the downregulation of collagen-II in chondrocytes, but IF pretreatment
inhibited IL-1β-induced downregulation of collagen-II (Fig. 6a). Con-
sistent with these observations, PCR analysis showed that mRNA ex-
pression of aggrecan and collagen-II decreased significantly after IL-1β
stimulation but recovered in chondrocytes pretreated with IF in a dose-
dependent manner (Fig. 6b, c). IF alone had no significant effects on
aggrecan and collagen-II expression in the chondrocytes (Supplemental
Fig. 4).
3.7. Effect of IF on NF-κB activation in human OA chondrocytes
To further explore the mechanism underlying the anti-inflammation
effect of IF, we focused on NF-κB signaling in chondrocytes. As shown
in Fig. 7a–c, IL-1β remarkably increased the phosphorylation of NF-κB
p65 and IκB-α in chondrocytes. In addition, IL-1β led to notable de-
gradation of IκB-α. In contrast, IF blocked IL-1β-stimulated

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phosphorylation of NF-κB p65 and IκB-α and the degradation of IκB-α.
Then we investigated the effect of IF on IL-1β-induced NF-κB p65
nuclear translocation by immunofluorescence staining. In chondrocytes
of control group the majority of p65 was located in the cytoplasm
(Fig. 7d). In chondrocytes stimulated by IL-1β, we observed enhanced
nuclear staining of p65 (Fig. 7e). However, IF inhibited IL-1β-induced
nuclear translocation of p65 in chondrocytes (Fig. 7f). IF alone had no
significant effects on IκB, p-IκB, p65, and p-p65 protein levels in the
chondrocytes (Supplemental Fig. 2).
4. Discussion
In this study we showed that IF decreased IL-1β-stimulated over-
production of PGE2, NO, COX-2, iNOS, MMP-1, MMP-3, MMP-13,
ADAMTS-4 and ADAMTS-5, while recovered IL-1β-stimulated down-
regulation of collagen-II and aggrecan in chondrocytes isolated from
OA patients, and these effects are correlated with the inhibition of IL-
1β-induced NF-κB activation by IF. These findings suggest that IF in-
hibits IL-1β-stimulated inflammatory response in OA progression.
Elevated IL-1β and TNF-α levels were found in synovial fluid of OA
patients [24,25]. In addition, IL-1β induced the expression of iNOS and
COX-2, which contributed to the production of NO and PGE2 [26].
Recent studies reported that NO increased the level of PGE2, which is
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Fig. 7. Effect of IF on NF-κB activation in
human OA chondrocytes. The protein levels
of p-p65, p65, p-IκB, IκB were determined
by Western blot analysis (a–c). The nuclear
translocation of p65 was detected by im-
munofluorescence staining. DAPI stained
the nuclei. In control chondrocytes, p65
was mainly localized in the cytoplasm (d).
Chondrocytes stimulated with IL-1β alone
demonstrated nuclear translocation of p65
(e), but was attenuated in chondrocytes
pretreated with IF (f). All experiments were
repeated three times. The values are
mean ± SD of three independent experi-
ments. #p < 0.05 compared with control
group. *p < 0.05, **p < 0.01 compared
with IL-1β group.
associated with the degeneration of articular cartilage by damaging
ECM and inhibiting the proliferation of chondrocytes [27,28]. Several
studies have indicated that the attenuation of inflammatory mediators
such as NO and PGE2 could weaken the progression of OA [29]. In this
study, we found that IF blocked the production of NO and PGE2, and
downregulated COX-2 and iNOS expression at both mRNA and protein
levels. Our results are consistent with previous study that IF blocked the
production of PGE2 and COX-2 in lipopolysaccharide (LPS) induced
acute lung injury in mice [30].
MMPs are considered as the main enzymes that regulate tissue re-
modeling and the degradation of ECM such as aggrecan and collagens
[31,32]. MMP-13 has been shown to target collagen-II which composes
the main structure of ECM [33]. Furthermore, ADAMTS members play
major role in cartilage degradation in OA, especially ADAMTS-4 and
ADAMTS-5, which are responsible for cleaving aggrecan [4,7]. There-
fore, targeting ADMATS-5 has been a major strategy in the treatment of
OA [34]. Using small interfering RNA (siRNA) to block ADAMTS-5
could restrain aggrecan loss from human OA cartilage explants [35]. In
this study, we found that IF inhibited IL-1β-stimulated MMP-1, MMP-3,
MMP-13, ADAMTS-4, and ADAMTS-5 production at both mRNA and
protein levels in human OA chondrocytes. Thus we speculated that IF
may exert anti-inflammation action by decreasing the production and
activation of MMPs and ADAMTS in the progression of OA.
J. Lin et al.
NF-κB signaling is a key pathway associated with OA pathogenesis
[36]. NF-κB is known to regulate the expression of inflammatory factors
including iNOS, COX-2 and MMPs [37]. Normally, NF-κB is located in
the cytoplasm combined with IκB, which is an inhibitory protein. Upon
stimulation by IL-1β, IκB would be phosphorylated and degraded, fol-
lowed by the dissociation of NF-κB p65 from IκB. NF-κB p65 would then
be translocated into the nucleus to activate inflammatory related genes
[38]. Previous studies showed that NF-κB inhibitor decreased the ex-
pression of MMP-3 and MMP-13 stimulated by IL-1β [39]. Additionally,
NF-κB p65 blocked the levels of COX-2, iNOS and MMP-9 in the de-
velopment of OA [40]. Therefore, targeted downregulation of NF-κB
might be effective for treating OA. In this study, we found that IF re-
markably blocked IL-1β-induced NF-κB activation and IκB-α degrada-
tion in OA chondrocytes. Furthermore, IF restrained nuclear translo-
cation of NF-κB p65 stimulated by IL-1β. Li et al. reported that IF
suppressed p65 translocation into the nucleus and inhibited iNOS
production in high-fat-induced hepatic lipid metabolism disorders in
mice [41]. In addition, IF inhibited TNF-α production and decreased
phospho-p38 in LPS-stimulated peritoneal macrophages [19]. Taken
together, our data suggest that IF inhibits NF-κB activation in the pro-
gression of OA. Nevertheless, further studies are needed to understand
the detailed mechanism by which IF inhibits NF-κB activation during
OA progression.
In conclusion, our study is the first to evaluate potential effects of IF
on human OA chondrocytes. Our results demonstrate that IF inhibits IL-
1β-induced ECM degradation and inflammation via the regulation of
NF-κB signaling, and suggest that IF may be a potential therapeutic
agent for OA.
Acknowledgements
The authors thank the staff of the Laboratory of the Orthopaedic
Research Institute and the Scientific Research Center of the Second
Affiliated Hospital of Wenzhou Medical University. This work was
supported by Zhejiang Province Public Welfare Technology Application
Research Project, China (No. 2015C33209).
Compliance with ethical standards
The study was in accordance with the Declaration of Helsinki and
Tokyo.
Conflict of interest
The authors declare that they have no conflict of interest.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.intimp.2018.09.003.
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