Biochemical and Biophysical Research Communications xxx (xxxx) xxx

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Biochemical and Biophysical Research Communications

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Vitexin alleviates non-alcoholic fatty liver disease by activating AMPK

in high fat diet fed mice
Shrirang Inamdar a, Ankita Joshi a, Sajad Malik a, Ramanamurthy Boppana b,
Saroj Ghaskadbi a, *
a
Department of Zoology, Savitribai Phule Pune University, Pune, 411007, India
b
National Centre for Cell Science, Pune, 411007, India

a r t i c l e i n f o a b s t r a c t

Article history: Non-alcoholic fatty liver disease (NAFLD) is a most common liver disorder characterized by accumulation
Received 21 August 2019 of fat in the liver and currently there is no approved treatment for it. Obesity and diabetes being leading
Accepted 25 August 2019 cause of NAFLD, compounds having anti-obesity activity and potential to reduce insulin resistance are
Available online xxx
considered suitable candidate for NAFLD treatment. In this study, we checked effect of vitexin, a naturally
occurring flavonoid, on high fat diet (HFD) induced NAFLD in C57BL/6J mice. In presence of vitexin,
Keywords:
significant reduction in body and liver weight, triglyceride and cholesterol content in serum and liver
Non alcoholic fatty liver disease (NAFLD)
was observed. Serum Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) levels were
Vitexin
AMP-activated protein kinase (AMPK)
reduced significantly by vitexin which were elevated in HFD group whereas serum lipase activity
Leptin receptor remained unchanged. Vitexin suppressed de novo lipogenesis by downregulating expression of Peroxi-
some proliferator-activated receptor g (PPARg), CCAAT/enhancer-binding protein-a (C/EBP-a), sterol
regulatory element-binding protein-1c (SREBP-1c), Fatty acid synthase (FAS) and Acetyl-CoA Carboxylase
(ACC). Additionally, it also enhanced fatty acid oxidation and lipolysis by upregulating Peroxisome
proliferator-activated receptor a (PPAR-a), carnitine palmitoyltransferase-1a (CPT-1a) and Adipose tri-
glyceride lipase (ATGL). Inhibition of lipogenesis and activation of lipolysis and fatty acid oxidation by
vitexin was found to be mediated by activation of AMP-activated protein kinase (AMPK). Vitexin also
improved insulin signalling by activating insulin receptor substrate-1 (IRS-1) and its downstream target
AKT. AMPK activation of vitexin was possibly through binding of vitexin to leptin receptor (LepR) which
was confirmed by molecular docking studies and by observed enhanced expression of LepR. Thus, we
propose that vitexin alleviates NAFLD by activating AMPK possibly by binding to LepR.
© 2019 Elsevier Inc. All rights reserved.

1. Introduction Since, there is no approved medical treatment for NAFLD,

NAFLD is characterized by the accumulation of fat (more than
5%) in the liver without a history of alcoholism or known liver
pathology. NAFLD has emerged as an important health problem
worldwide with prevalence of around 25% [1] whereas, in India the
prevalence is around 32% [2]. NAFLD includes steatosis and stea-
tohepatitis which can progress further to fibrosis, cirrhosis and
even to hepatocarcinoma [3]. Obesity and diabetes are a major risk
factors leading to NAFLD and their increased prevalence due to
modern dietary habits and sedentary lifestyle has resulted in
increased NAFLD prevalence.
* Corresponding author.
E-mail addresses: ssg@unipune.ac.in, sghaskadbi@gmail.com (S. Ghaskadbi).
compounds which control obesity would prove effective in man-
agement of NAFLD [4]. Vitexin, (apigenin-8-C-D-glucopyranoside),
is a natural flavonoid found in certain herbs such as hawthorn herb,
mung bean and fenugreek. It has wide range of pharmacological
uses, including anti-obesity, anti-inflammatory, antioxidant and
anti-tumor [5]. Vitexin and extract containing vitexin have been
shown to be effective in reduction of diet induced obesity [6e9]
and thus would be a promising candidate for control of NAFLD.
High fat diet (HFD) induces NAFLD as consequence of imbalance
between energy intake and its expenditure. Leptin and LepR
regulate the body weight by balancing food intake and energy
expenditure by activating AMPK [10]. AMPK is a major energy
sensor of the cell and regulates hepatic and adipose lipid meta-
bolism by modulating lipogenesis, lipolysis, gluconeogenesis and
adipogenesis [11]. AMPK is known to inhibit de novo lipogenesis by

https://doi.org/10.1016/j.bbrc.2019.08.139
0006-291X/© 2019 Elsevier Inc. All rights reserved.

Please cite this article as: S. Inamdar et al., Vitexin alleviates non-alcoholic fatty liver disease by activating AMPK in high fat diet fed mice,
Biochemical and Biophysical Research Communications, https://doi.org/10.1016/j.bbrc.2019.08.139
2 S. Inamdar et al. / Biochemical and Biophysical Research Communications xxx (xxxx) xxx
down regulating key lipogenic genes such PPARg, C/EBP-a and
SREBP-1c in HFD fed animals [11,12]. In addition, AMPK is also
known to promote fatty-acid oxidation by suppressing ACC and FAS
activity [13]. It also up-regulates expression of CPT-1a, a key
regulator of fatty-acid oxidation [12]. Vitexin is known to activate
AMPK signaling during alleviating obesity [8] and endothelial
injury [14]. In addition to alterations in energy homeostasis, HFD is
also known to induce insulin resistance which in turn leads to
NAFLD [15]. Hence, compounds improving both energy homeo-
stasis and insulin signaling would be advantageous for the
treatment.
This study was undertaken to check effect of vitexin on diet
induced NAFLD in mice and deduce the mechanism behind this. We
confirmed that vitexin inhibits NAFLD by activating AMPK which in
turn inhibits lipogenesis and activates lipolysis and fatty acid
oxidation. In addition, vitexin also improved insulin signaling in
liver of HFD fed animals. We propose that the vitexin activates
AMPK by probably binding and activating LepR.
2. Materials and methods
2.1. Animal studies
C57BL/6J mice used in the study were maintained at Experi-
mental Animal Facility (EAF) at National Centre for Cell Science
(NCCS), Pune, India. Animals were housed at 12 h light/dark cycle
with ad libitum access to water and food. All animal experiments
were carried out as per the requirement and guidelines of the
Committee for the Purpose of Control and Supervision of Experi-
ments on Animals (CPCSEA), Government of India, and after
obtaining permission of the Institutional Animal Ethics Committee
(IAEC/2018/B-332).
Total 30 animals (6e8 weeks old) were used in the study and
were divided equally into 5 groups namely Control (ND), High fat
diet (HFD), HFD supplemented with 1 mg/kg vitexin (HFDþ 1 mg
Vit), with 10 mg/kg vitexin (HFDþ 10 mg Vit) and with 20 mg/kg
vitexin (HFDþ 20 mg Vit). ND group animals were fed with normal
diet (5% kcal fat) purchased from Nutrivet Life Sciences Pune, India
whereas, HFD comprising 60% kcal fat (D12492, Research diet Inc.,
USA) was given to all other groups for eight weeks. Vitexin was
given daily by oral gavage. Body weight of all mice was monitored
weekly. At the end of 8 weeks, blood was collected from orbital
plexus and mice were sacrificed by cervical dislocation. Liver and
adipose tissue were harvested and snap frozen in liquid N2. Part of
the tissue was fixed in 4% paraformaldehyde and used for histo-
logical analysis. Remaining tissue was stored at 80 OC and pro-
cessed for protein and RNA isolation.
2.2. Biochemical analysis
Total cholesterol, triglycerides, ALT, AST and lipase activity were
measured using commercially available kits (Accurex Biomedical
PVT LTD, India) following manufacturer's protocol.
2.3. Histochemical analysis
Liver tissue was fixed in 4% paraformaldehyde and 10 mm sec-
tions were collected on poly L-lysine coated slides using cryotome
(Leika CM 1250, Germany) and stained with oil red O. Images were
captured using Nikon eclipse TiS (Nikon, Japan) microscope.
2.4. RNA isolation and quantitative real time PCR (qRT-PCR)
RNA from liver tissue was isolated using Trizol reagent, quan-
titated using Nanophotometer (Eppendorf, Germany) and purity of
Please cite this article as: S. Inamdar et al., Vitexin alleviates non-alcoholic fatty liver disease by activating AMPK in high fat diet fed mice,
Biochemical and Biophysical Research Communications, https://doi.org/10.1016/j.bbrc.2019.08.139
RNA was assessed by calculating A260/280. cDNA was prepared using
Mulv-Reverse transciptase (New England Biolabs, USA) and qRT-
PCR was performed on Step One Plus (Applied Biosystems, USA)
using SYBR green and gene specific primers (Supplementary
Table 1) for PPARg, PPARa, C/EBP-a, SREBP-1c, CPT-1a, PGC-1a,
and ATGL. Fold-change in expression was calculated by using 2-DDCT
method where b-Actin was used as a house-keeping gene.
2.5. Western blot analysis
Liver tissue samples were homogenized in the lysis buffer
containing 20 mM Tris, pH 7.5, 1 mM EDTA (pH 8.0), 150 mM NaCl,
1% Triton X-100, 10% glycerol, 1 mM phenylmethylsulfonyl fluoride;
1 mM sodium orthovanadate, 0.5% sodium deoxycholate, 10 mM
sodium fluoride, 1 mM DTT, 12.5 mM b-glycerophosphate and 1X
Protease inhibitor cocktail and equal amount of protein was loaded
on 12% SDS-PAGE. Proteins were electro transferred to PVDF
membrane (Millipore, USA) at 80 mA constant current. Blots were
then saturated with BSA, reacted with different primary antibodies
(PPARg, C/EBP-a, FAS, ACC, p-AMPK, t-AMPK, p-AKT, t-AKT, p-IRS1,
t-IRS1 and b-Actin procured from Elabscience, USA) and then
incubated with secondary anti-rabbit antibody (Sigma, USA) tagged
to horseradish peroxidase. Signals were detected by chem-
iluminescence using luminol (Advansta, USA) as a substrate and
documented using chemidoc system (Biorad, USA).
2.6. Molecular modeling and docking studies
Sequence of mouse LepR obtained from Uniprot database (www.
uniprot.org) was blasted against protein data bank (PDB) using
BLASTp. Human LepR crystal structure was used as a template to
generate homology model of mouse LepR using SWISS-MODEL
server (https://swissmodel.expasy.org/). Model was selected
based on QMEAN score and used further for docking studies. For
this, structure of vitexin and structure of mouse LepR constructed
using Swiss model was submitted to AutoDock Vina software and
results obtained were visualized and analyzed using Chimera
software [16].
2.7. Statistical analyses
Each experiment was performed at least three times and the
data are shown as mean with standard error (SE). Statistical sig-
nificance was calculated by SPSS 19.0 software with one-way
ANOVA. P-value<0.05 was considered statistically significant.
#,
*p < 0.05, ##,**p < 0.001, ###,***p < 0.0001.
3. Results
3.1. Vitexin reduces body weight gain and tissue weight in HFD-
induced obese mice
Significant increase in body and liver weight was observed in
HFD group animals after 8 weeks compared to those of ND group
animals (Fig. 1A). Vitexin treatment significantly reduced the body
weight gain induced by HFD in a dose dependent manner (Fig. 1A)
concomitant with decreased liver weight (Supplementary Table 2).
Significantly elevated levels of serum total cholesterol and tri-
glycerides were found in HFD group animals compared to ND. This
increase was significantly reduced by vitexin in a dose dependent
manner (Fig. 1B and C). Both AST and ALT levels found to be
increased significantly in HFD fed mice were reduced after vitexin
treatment at 1 and 10 mg/kg body weight and remained unchanged
at higher concentration (Fig. 1D and E). Intestinal lipase activity was
not found to be altered in mice treated with vitexin, indicating that
 S. Inamdar et al. / Biochemical and Biophysical Research Communications xxx (xxxx) xxx 3

Fig. 1. Vitexin attenuates NAFLD by reducing body weight gain and normalizing different serum parameters. Effect of vitexin on body weight (A), total cholesterol (B), total
triglyceride (C) ALT (D) AST (E) and lipase activity (F) after 8 weeks. (G) Liver sections of mice from different groups stained with H & E. (H) Estimation of triglyceride in liver of mice
from different groups. # represents significance level with respect to ND group. * represents significance level with respect to HFD group. #,*p < 0.05, ##,**p < 0.001,
###,
***p < 0.0001.

the effect of vitexin is not mediated by preventing dietary lipid
uptake (Fig. 1F).
3.2. Vitexin inhibits development of NAFLD
Enhanced accumulation of lipids in liver, a characteristic feature
of NAFLD, was confirmed by oil red O staining in liver sections of
HFD fed animals and this was significantly rescued in liver of mice
treated with vitexin in a dose dependent manner (Fig. 1G). Simi-
larly, significant increase in triglyceride accumulation in liver of
HFD group mice was inhibited by treatment with vitexin (Fig. 1H).
suppressed in liver of HFD-induced obese mice while it was
significantly increased in presence of vitexin in a dose dependent
manner thereby promoting lipolysis (Fig. 2B). PGC-1a is a master
regulator of mitochondrial biogenesis and oxidative metabolism
and is known to activate expression of CPT-1a involved in transport
of fatty acids into mitochondria. Expression of both PGC-1a and
CPT-1a was significantly decreased in liver of HFD fed mice and
vitexin increased their expression in a dose dependent manner
(Fig. 2B).
3.4. Vitexin alleviates inhibition of insulin signaling

3.3. Vitexin alleviates NAFLD state of liver by down regulating
lipogenesis and by up regulating lipolysis and fatty acid oxidation
PPARg and C/EBP-a, are important transcription factors pro-
moting lipogenesis and their expression was indeed found to be
significantly increased both at RNA (Fig. 2A) and protein level
(Fig. 2C and D) in liver of HFD group and vitexin suppressed their
expression in a dose dependent manner. Expression of SREBP-1c
(Fig. 2A) which is activated by PPARg and its downstream targets
FAS and ACC, rate limiting enzymes of de novo lipogenesis, was
found to be significantly increased in liver of HFD fed animals and
vitexin reduced it in a dose dependent manner (Fig. 2C and D).
Expression of ATGL, gene involved in lipolysis and transcription
factor PPARa was checked by qRT-PCR and was found to be
NAFLD is associated with impaired insulin signaling [17] due to
enhanced inhibitory phosphorylation at Ser-307 residue of IRS-1.
We indeed, observed significantly increased levels of p-IRS-1(Ser-
307) in liver of HFD fed animals and vitexin reduced this inhibition
in a dose dependent manner. (Fig. 3A and B). We also checked
activation of AKT (phosphorylation levels of Ser-473), a down-
stream target of IRS-1 and found it significantly reduced in HFD fed
mice. Vitexin reversed this effect in a dose dependent manner
(Fig. 3A and B).
3.5. Vitexin activates AMPK possibly through LepR
AMPK is a master regulator of energy homeostasis and is known
to modulate both lipogenesis by inhibiting SREBP-1c and lipolysis

Please cite this article as: S. Inamdar et al., Vitexin alleviates non-alcoholic fatty liver disease by activating AMPK in high fat diet fed mice,
Biochemical and Biophysical Research Communications, https://doi.org/10.1016/j.bbrc.2019.08.139
4 S. Inamdar et al. / Biochemical and Biophysical Research Communications xxx (xxxx) xxx

Fig. 2. Vitexin alleviates NAFLD by inhibiting lipogenesis and by promoting lipolysis and fatty acid oxidation in liver. Relative mRNA expression of genes in lipogenesis (A), and
lipolysis and FFA oxidation (B) in liver of mice from different groups. (C) Expression of protein involved in lipogenesis in liver of mice from different groups and (D) densitometric
analysis of western blot in fig. (C). # represents significance level with respect to ND group. * represents significance level with respect to HFD group. #,*p < 0.05, ##,**p < 0.001,
###,
***p < 0.0001.

by activating PGC-1a and PPARa. We found decreased p-AMPK in 4. Discussion

liver of HFD fed animals while in case of vitexin treated mice this
decrease was inhibited in a dose dependent manner (Fig. 3A and B).
AMPK is known to be activated by LepR [10]. Therefore, in order
to determine whether vitexin would also activate AMPK by binding
to LepR, molecular docking studies were performed. Since crystal
structure of mouse LepR is not available, a blast was performed
using mouse LepR sequence and PDB database wherein mouse
LepR showed maximum similarity (86.87%) with human LepR (PDB
id 3V6O) (Fig. 3C). Therefore, crystal structure of human LepR was
used as a template for generation of homology model of mouse
LepR using Swiss model server. The model with the best QMEAN
score was selected (QMEAN score ¼ 1.5), validated by Ramchan-
dran plot and was used for further study (Supplementary Fig. 1).
Superimposing human and homology model generated mouse
LepR structure showed good match with RMSD value of 0.096 Ao
(Fig. 3D). Vitexin showed docking at the leptin binding site pocket
of the LepR with vena score of 5.9 indicating strong affinity.
Vitexin formed hydrogen bonds with the LYS-484 and ASN-485
amino acids from the active site region which lie in binding re-
gion of leptin with its receptor (Fig. 3E and F). We also performed
docking studies with LepR inhibitor curcumin and observed that
the inhibitor binds strongly to LepR at a site similar to that of
vitexin with vena score of 6.5. Similar amino acids are involved in
the formation of H bonds as that of vitexin (Fig. 3G). To further
confirm we checked expression of LepR by qRT-PCR in presence of
vitexin and indeed found it to be activated (Fig. 3H). Based on this
and in silico analysis, vitexin seems to activate LepR which further
activates AMPK.
In the current study, we demonstrate the potential of vitexin to
inhibit liver steatosis observed in NAFLD in mice fed with HFD for
eight weeks. Obesity induced NAFLD is associated with increased
body and liver weight, enhanced serum triglyceride and cholesterol
level along with accumulation of fat in liver. This was confirmed by
staining liver tissue sections by lipid specific stain, oil red O and by
estimating the triglyceride and cholesterol levels in liver and
serum. HT048, a combination of Crataegus pinnatifida leaf and Cit-
rus unshiu peel extracts containing vitexin has been shown to
reduce body and liver weight and serum levels of triglyceride and
cholesterol in HFD fed rat [6]. Similarly, ethanol extract of mung
bean containing vitexin, significantly reduced the body weight and
serum lipid levels in KK-Ay mice when challenged with HFD [7].
More recently Peng et al. [8] and Seyedan et al. [9] have reported
that vitexin or extract of Cynometra cauliflora containing vitexin
reduced body and liver weight and serum lipid levels in HFD fed
mice similar to our observations.
Serum lipase hydrolyzes dietary fats into triglycerides and free
fatty acids which are then absorbed in the intestine. Commercially
available anti-obesity drugs such as orlistat and resveratrol are
known to exert their effect by inhibiting pancreatic lipase activity.
We did not find alteration in the serum lipase activity at all con-
centrations of vitexin used indicating that effect of vitexin is not
mediated by inhibiting digestion of fat and its intestinal absorption.
Increased triglyceride accumulation in liver can be either due to
increased FFA uptake and de novo lipogenesis in liver and/or
decreased oxidation of stored fatty acids. FAS and ACC are the

Please cite this article as: S. Inamdar et al., Vitexin alleviates non-alcoholic fatty liver disease by activating AMPK in high fat diet fed mice,
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 S. Inamdar et al. / Biochemical and Biophysical Research Communications xxx (xxxx) xxx 5

Fig. 3. Vitexin alleviates inhibition of insulin signaling and activates AMPK possibly through LepR. (A) Protein expression of p-IRS-1, t-IRS1 p-AKT t-AKT and p-AMPK and t-
AMPK in liver of mice from different groups (B) Densitometric analysis of western blot in (a) (C) Pairwise sequence alignment of human and mouse LepR sequence. (D) Super-
imposed crystal structures of human and mouse LepR. Binding of vitexin (shown in green in E and blue in F) to LepR in hydrophobic surface (E) and ribbon (F) representation.
Hydrogen bonds are represented in red. (G) Binding of antagonist curcumin (shown in blue) to LepR. Hydrogen bonds are represented in red. # represents significance level with
respect to ND group. * represents significance level with respect to HFD group. #,*p < 0.05, ##,**p < 0.001, ###,***p < 0.0001. (For interpretation of the references to colour in this
figure legend, the reader is referred to the Web version of this article.)

important enzymes in de novo lipogenesis which are regulated by
transcription factor, SREBP-1c, which in turn is activated by PPARg
and C/EBP-a [18]. We found that vitexin inhibited expression of all
these genes in a dose dependent manner. Herbal formulation
HT048 containing vitexin has also been shown to inhibit lipogen-
esis in 3T3-L1 pre-adipocytes and HFD fed rats by inhibiting
expression of PPARg, C/EBP-a and FAS [6]. Similarly, mung bean
ethanol extract containing vitexin suppressed the lipogenic genes
PPARg, C/EBP-a and ACC in muscle of KK-Ay diabetic mice [7].
Recently, Peng et al. [8] have shown the potential of vitexin in
reducing the expression of C/EBP-a and ACC in adipose tissue in
HFD fed mice. We then checked the expression of lipolysis and fatty
acid oxidation related genes in liver. PPARa and PGC-1a govern the
fatty acid oxidation by enhancing the expression of ATGL, a rate
limiting enzyme in lipolysis, and CPT-1a, which is essential for
transport of long-chain fatty acids into mitochondria [19]. Results
revealed that vitexin indeed increased expression of these genes in
a dose dependent manner in liver suggesting that vitexin mediates
its effect also by activating lipolysis and subsequent fatty acid
oxidation.
AMPK is a master regulator of energy homeostasis and known to
play role in pathogenesis of hepatic steatosis [20]. AMPK activation
inhibits lipogenesis by suppressing the expression of C/EBP-a,
SREBP-1c, FAS as well as ACC and enhances lipolysis and b-oxida-
tion by activating the expression of PGC-1a, ATGL and CPT-1a [21].
We found that in presence of vitexin AMPK was activated signifi-
cantly in a dose dependent manner suggesting the effect of vitexin
is mediated by activation of AMPK signaling.
In addition to altered lipogenesis and lipolysis, impaired insulin
signaling due to development of hepatic insulin resistance is very
closely linked to NAFLD [17,22]. Different plant extracts with po-
tential to improve insulin sensitivity and activation of downstream
AKT have shown promising therapeutic effects in NAFLD [17,23,24].
In this study, we also observed that vitexin improved insulin
signaling by inhibiting Ser-307 phosphorylation of IRS-1 and acti-
vating AKT; a master regulator of Insulin signaling.
LepR is known to activate AMPK by activating Calcium/
calmodulin-dependent protein kinase kinase 2 (CaMKK2) and
IRS-1 via JAK/STAT3 pathway [10]. We therefore, speculated that
vitexin may govern its action by activating LepR. To confirm this,
initially we performed docking studies. Since, the crystal structure
of mouse LepR is not known, we generated and validated homology
model using human LepR crystal structure as a template [25]. The
LepR (LR), encoded by db gene contains two cytokine receptor

Please cite this article as: S. Inamdar et al., Vitexin alleviates non-alcoholic fatty liver disease by activating AMPK in high fat diet fed mice,
Biochemical and Biophysical Research Communications, https://doi.org/10.1016/j.bbrc.2019.08.139
6 S. Inamdar et al. / Biochemical and Biophysical Research Communications xxx (xxxx) xxx

Fig. 4. Schematic diagram showing mechanism of action of protective effects of vitexin against NAFLD. Vitexin binds to LepR leading to activation of AMPK. Phosphorylated
AMPK activates PGC-1a, PPARa and ATGL leading to increased lipolysis and fatty acid oxidation. And inhibits PPARg, SREBP-1c, ACC and FAS leading to inhibition of de novo
lipogenesis. Vitexin also improves insulin signaling by activating IRS-1 leading to activation of AKT.

homology (CRH) modules. CRH2 domain which is composed of two Acknowledgements

sub-domains with fibronectin type III folds (residues 428e535 and
536e635, respectively in human) was identified as the main high-
affinity binding site for leptin on the LepR [26,27]. Blast studies and
earlier reports confirm that this region is highly conserved in
mouse [28]. Our docking studies revealed that vitexin binds
strongly at CRH2 domain of mouse LepR i.e. the leptin binding re-
gion. In addition, we found that binding site of LepR inhibitor
curcumin overlaps with that of vitexin. We also observed increased
expression of LepR in presence of vitexin by q-RT PCR. Thus,
docking studies and enhanced LepR expression suggest that the
effect of vitexin is probably governed by activation of LepR. How-
ever, this needs to be confirmed by further experimentation. Leptin
bound at CRH2 domain also interacts with Ig like domain of the
second LepR via binding site-III thereby inducing 2:4 leptin:LepR
complex and receptor activation [10]. It would be interesting to see
whether vitexin acts in a similar manner.
In summary, present study provides evidence to support the
potential therapeutic effect of vitexin on reducing liver steatosis in
HFD fed mice. This observed effect is attributed to enhanced fatty
acid oxidation and lipolysis in addition to anti-lipogenesis activity.
Vitexin was also found to enhance insulin signaling in the liver.
These effects are achieved by activating the AMPK signaling
possibly by binding and activating LepR (Fig. 4).
Conflicts of interest
Authors declare that there is no conflict of interest.
Authors acknowledge financial support from Department of
Science and Technology - Promotion of University Research and
Scientific Excellence (DST-PURSE, GOI-A-670) and University Grant
Commission-Career Advancement Scheme (UGCCAS, F-5-2/
2005(SAP-II) program of the Department of Zoology, Savitribai
Phule Pune University, India and SERB project (GOI-A-752) to SG.
Fellowship provided by Dr. D. S. Kothari fellowship from UGC to SI
and DST-PURSE fellowship to SM is duly acknowledged. Authors
thank Dr. Abhijit Kulkarni, Department of Bioinformatics, SPPU and
Dr. Susmit Sambhare,ACTREC, Mumbai for their help in bioinfor-
matics related work.
Appendix A. Supplementary data
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.bbrc.2019.08.139.
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