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Journal of Future Foods 2-1 (2022) 61–68

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Effect of high quality dietary fiber of Hericium erinaceus on

Effect of high
lowering bloodquality
lipid indietary fiber of Hericium
hyperlipidemia mice erinaceus on
lowering blood
a,b lipid in
a,c hyperlipidemia
a,b micea,b,*
Tingting Liu , Nan Wang , Xinle Xu , Dawei Wang
Tingting
a Liua,b, Nan Wanga,c, Xinle Xua,b, Dawei Wanga,b,*
School of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China
b
a Engineering
School Research
of Food ScienceCenter of Grain Deep-processing
and Engineering, andUniversity,
Jilin Agricultural High-effeciency Utilization
Changchun of Jilin
130118, ChinaProvince, Changchun 130118, China
c
b Key Laboratory
Engineering of Technological
Research Innovations
Center of Grain for Grain Deep-processing
Deep-processing andUtilization
and High-effeciency High-effeciency Utilization
of Jilin Province,ofChangchun
By-products of JilinChina
130118, Province, Changchun 130118, China
c
Key Laboratory of Technological Innovations for Grain Deep-processing and High-effeciency Utilization of By-products of Jilin Province, Changchun 130118, China

ARTICLE INFO ABSTRACT

ARTICLE INFO ABSTRACT
Article history: The purpose of this work is to describe the structure and hypoglycaemic activity of high-quality Hericium
Received 28 August 2021
Article history: erinaceus
The purpose dietary
of thisfiber
work(HQ-HDF), andthe
is to describe compared
structurewith
and the H. erinaceusactivity
hypoglycaemic dietaryoffiber (HDF). Scanning
high-quality Hericium
Received
Received in28revised
Augustform
202112 September 2021 electron
erinaceusmicroscopy and(HQ-HDF),
dietary fiber particle sizeand
distribution
compared showed
with thethatH.
theerinaceus
ultrasonic-microwave
dietary fiber assisted
(HDF). enzymatic
Scanning
Accepted 10revised
Januaryform
202212 September 2021 method
Received in
Available Online 25 March electron signifi cantly and
microscopy changed the size
particle surface structure showed
distribution of HQ-HDF andultrasonic-microwave
that the reduced the particle size. The enzymatic
assisted results of
Accepted 10 January 2022 2022 high-performance
method significantly gelchanged
chromatography
the surfaceshowed that
structure of the ultrasonic-microwave
HQ-HDF and reduced the assisted enzymatic
particle size. method
The results of
Available Online 25 March 2022 made the macromolecular
high-performance components degrade
gel chromatography showedintothatsmall molecular components.assisted
the ultrasonic-microwave HQ-HDF has signifi
enzymatic cantly
method
Keywords: improved hyperlipidemic activity in vitro. The into
results showed that HQ-HDF has HQ-HDF
blood lipid-lowering. The
Hericium erinaceus made the macromolecular components degrade small molecular components. has significantly
Keywords: levels
improvedof total cholesterol,activity
hyperlipidemic triglycerides,
in vitro.low-density
The results lipoprotein
showed thatand high-density
HQ-HDF lipoprotein
has blood in serumThe
lipid-lowering. of
Dietary
Hericiumfiber
erinaceus high-fat
Structure characterization levels of mice in medium triglycerides,
total cholesterol, and high dose group were
low-density significantly
lipoprotein increased. High
and high-density dose ofin HQ-HDF
lipoprotein serum of
Dietary fiber signifi cantly
high-fat micedecreased
in mediumthe activities
and highofdoseglutamic-pyruvic
group were transaminase
significantlyand glutamicHigh
increased. oxalacetic
dose transaminase,
of HQ-HDF
High-fat mice
Structure characterization increased thedecreased
activitiesthe
of superoxide
Hypolipidemic significantly activities of dismutase and glutathione
glutamic-pyruvic peroxidase.
transaminase High dose
and glutamic of HQ-HDF
oxalacetic could
transaminase,
High-fat mice decreased the content
increased the activitiesofof
malonaldehyde. In addition,
superoxide dismutase andhistopathological observation
glutathione peroxidase. Highofdose
miceofliver showedcould
HQ-HDF that
Hypolipidemic HQ-HDF
decreased could improve
the content of the lipid metabolism
malonaldehyde. disorderhistopathological
In addition, of liver cells. observation of mice liver showed that
HQ-HDF could© 2022 Beijingthe
improve Academy of Food Sciences.
lipid metabolism disorder ofPublishing services by Elsevier B.V. on behalf of KeAi
liver cells.
©©2022
2022Beijing
Beijing Academyof
Communications
Academy of Food
Co.,
FoodLtd.Sciences.
This is an
Sciences. Publishing
open access
Publishing services by
articleby
services Elsevier
under the CC
Elsevier B.V. on behalf
behalf of
BY-NC-ND
B.V. on of KeAi
license
KeAi
Communications Co., Ltd. This is an (http://creativecommons.org/licenses/by-nc-nd/4.0/).
open access article under the CC BY-NC-ND license
Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
(http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction polysaccharides and purified polysaccharides of H. erinaceus could

1. Introduction relieve constipation
polysaccharides and and
purifimaintain of H. erinaceus
intestinal function.
ed polysaccharides Cui et al. [9]
could
Hericium erinaceus is a precious fungus in both food and discovered a new glycoprotein
relieve constipation and maintain HEG-5 in H.function.
intestinal erinaceus,Cuiwhich
et al. had
[9]
Hericium
medicine [1]. Wild H. erinaceus
erinaceus is distributed
is a precious fungusin in
East Asia,
both Europe
food and antitumor
discoveredand hemagglutination
a new activities.
glycoprotein HEG-5 H. erinaceus,
in Among them, the content
which had
and China,[1].
medicine Wildit H.
where erinaceus
mainly grows isondistributed
dead or dying wood.
in East Numerous
Asia, Europe of HDF can
antitumor andreach 34%. β-Glucanactivities.
hemagglutination and chitin are thethem,
Among most the
researched
content
studies
and China,havewhere
shown that it grows
it mainly containson 44.24%
dead or carbohydrates, 25.45%
dying wood. Numerous of HDF
and usedcan in H. 34%.
DFreach erinaceus.
β-Glucan
Xie etandal.chitin
[10] extracted β-glucan
are the most from
researched
protein, 6.44%
studies have fat and
shown thatvarious minerals,
it contains 44.24%etc.carbohydrates,
[2]. Modern medicine
25.45% the
andfruited
used DF of H.
in H.
body erinaceus,
erinaceus. Xieand
et structural analysis β-glucan
al. [10] extracted showed thatfrom it
has proved
protein, that H.
6.44% faterinaceus
and variouscan minerals,
enhance immunity [3], antineoplastic
etc. [2]. Modern medicine hadfruited
the 1→3 and body1→6of H. erinaceus,with
connection and astructural
molecular weightshowed
analysis of 13.3that
kDa. it
activity
has proved[4],that H. erinaceus
oxidation resistance [5], decrease
can enhance blood[3],
immunity glucose [6] and
antineoplastic Feng
had 1→3 et al.
and[11]
1→6extracted
connection high
withmolecular
a molecular weightβ-glucan
weight and
of 13.3 kDa.
blood
activitylipid
[4], [7],
oxidation resistance H.
etc. Therefore, [5],erinaceus
decrease has
bloodbeen favoured
glucose by
[6] and founded
Feng et that hydrogen
al. [11] bonds high
extracted drivedmolecular
the interaction
weightbetween β-glucan
β-glucan and
more
blood and
lipidmore
[7], consumers. Wang
etc. Therefore, H.eterinaceus
al. [8] founded thatfavoured
has been both crude
by and starch
founded thatgranules,
hydrogenthus inhibiting
bonds drived thetheinteraction of wheatβ-glucan
digestion between starch.
more and more consumers. Wang et al. [8] founded that both crude Liao et al. granules,
and starch [12,13] prepared hydrogels
thus inhibiting with Hericium
the digestion erinaceus
of wheat starch.
*
Correspondence author at: School of Food Science and Engineering, Jilin Agricultural
polysaccharides,
Liao et al. [12,13] and the results
prepared showedwith
hydrogels thatHericium
hydrogel erinaceus
had good
* University,
Correspondence author at: School Changchun
of Food Science130118 China.
and Engineering, Jilin Agricultural
elasticity and lightand
polysaccharides, transmittance.
the resultsHowever,
showed that therehydrogel
are few systematic
had good
E-mail address: wangdawei@jlau.edu.cn (D.W Wang)
University, Changchun 130118 China. studies onand
elasticity HDF. light transmittance. However, there are few systematic
Peer review
E-mail under
address: responsibility of KeAi (D.W
wangdawei@jlau.edu.cn Communications
Wang) Co., Ltd. Hyperlipidemia
studies on HDF. relates to a disease in which the blood lipid
Peer review under responsibility of KeAi Communications Co., Ltd. levelHyperlipidemia
is too high andrelatesthe lipid
to metabolism
a disease iniswhichdisordered. The main
the blood lipid
level is too high and the lipid metabolism is disordered. The main
Publishing services by Elsevier
Publishing services by Elsevier
http://doi.org/10.1016/j.jfutfo.2022.03.018
2772-5669/© 2022 Beijing Academy of Food Sciences. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC
http://doi.org/10.1016/j.jfutfo.2022.03.018
BY-NC-ND
2772-5669/©license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
2022 Beijing Academy of Food Sciences. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC
10.1016/j.jfutfo.2022.03.018
BY-NC-ND
2772-5669license
© 2022(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Beijing Academy of Food Sciences. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the
CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
62 Tingting Liu et al. / Journal of Future Foods 2-1 (2022) 61–68
62 T.T. Liu et al. / Journal of Future Foods 2 (2022) 61-68
manifestations are the abnormal increase of the TC or TG or LDL-C
content in the blood or the abnormal decrease of HDL-C content [14].
Recent epidemiological studies have found that DF has a function
to improve obesity and hyperlipidemia [15]. DF consists of
carbohydrate polymers that cannot be used in the small intestine, but
can be partially fermented by intestinal microbes after entering the
large intestine [16]. So DF is known as the seventh nutrient in the
body. According to solubility, DF can be divided into water-soluble
dietary fiber (SDF) and water-insoluble dietary fiber (IDF). SDF
includes pectin and some hemicellulose, while IDF consists of
cellulose, hemicellulose and lignin [17]. SDF is an important aspect
of DF because it has better physical and chemical properties and
physiological functions [18,19]. Many scholars have believed that the
content of SDF in high quality dietary fiber should be greater than
10% [20]. The mechanism of dietary fiber lowering blood lipids may
be as follows: first, DF cannot be digested in the gastrointestinal tract,
which leads to a decrease in satiety and energy intake; secondly, DF
in the gastrointestinal tract may interfere with lipid absorption due to
the binding ability of DF; thirdly, DF binds cholesterol and bile acids,
and indirectly reduces blood lipid levels due to the emulsification of
bile acids.
Ultrasound-microwave-assisted enzymatic method was used
to prepare HQ-HDF and analyzed the structural properties and
hypolipidemic activity in this study. It is a nourishing food with
lipid-lowering function. This paper provides theoretical basis for the
development of lipid-lowering functional health food.
2. Materials and methods
2.1 Materials and reagents
Residue of H. erinaceus (HER) made in the lab. Cellulase was
purchased from Sigma-Aldrich (St. Louis, MO, USA). BALB-c
mice (age 6−8 weeks) were purchased from Beijing Weitong Lihua
Laboratory Animal Science and Technology Co., Ltd. (Beijing,
China). The commercial kit of TC, TG, LDL-C, HDL-C, SOD, MDA
and GSH-Px was obtained from Nanjing Biotechnology Co. Ltd.
(Nanjing, China).
2.2 Preparation of HDF
We refered to GB 5009.88-2014 Determination of Dietary Fiber
in Food for experiment. The HER (5.0 g) were dissolved in deionized
water (200 mL), and then added to alkaline protease for enzymatic
hydrolysis. The enzymolysis solution was inactivated in boiling water
and precipitated by anhydrous ethanol. The precipitate was dried. The
obtained precipitate was labeled as HDF.
2.3 Preparation of HQ-HDF
First, HER (5.0 g) were dissolved in deionized water (200 mL).
Then, we adjusted the pH of the solution to 5.5. 3% Celluloses
were added to the solution, and ultrasonic processing (1.50 W/mL)
was continued for 75 min at 55 °C. The solution was boiled to stop
the enzyme, and flocculating alcohol precipitation overnight. The
precipitate was freeze-dried. The obtained precipitate was labeled as
HQ-HDF.
2.4 Structure of HQ-HDF
2.4.1 Scanning electron microscopy
The HDF and HQ-HDF were fixed with conductive adhesive for
N2 purging. And then they were observed and analyzed by scanning
electron microscopy (Phenom Pro) with ×200, ×1 000, ×5 000
magnification after gold plating. The accelerating voltage was 5 kV.
The probe mode was backscatter.
2.4.2 Particle size distribution
The particle size distribution of the HDF and HQ-HDF was
given in laser particle size analyzer (Malvern Mastersizer 3000E).
Test conditions: 0.1% DF suspension was in addition to the flowing
distilled water one by one, and when the shading rate reached 9%, the
measurement was clicked. The refractive indices of DF and water was
1.473 and 1.330, respectively.
2.4.3 SDF relative molecular mass determination
SDF was extracted from HDF and HQ-HDF. The relative
molecular weight of SDF was determined by high performance
gel permeation chromatography (1515). The SDF (2 mg/mL)
was dissolved in ultrapure water. The solution was filtered by a
0.45 μm microporous membrane. The standard curves were plotted with
Dextran T standard (molecular weight: 10, 40, 70, 500 and 2 000 kDa).
2.5 In vitro hypoglycemic capacities of HQ-HDF
2.5.1 Determination of inhibition ability of pancreatic lipase
Methods refered to Benitez et al. [22]. Among them: A is the
control group; a is the control blank group; B is the experimental
group of samples; b is the blank sample group.
B−b
Inhibition rate (%) = (1 − ) × 100 (1)
A−a
2.5.2 Determination of cholesterol binding capacity
Methods refered to Wang et al. [23]. The cholesterol content was
detected by OPA method. Standard cholesterol curve: y = 0.012x +
0.049 9, R2 = 0.998 4. Among them: A0 is the cholesterol content in
the egg yolk; A1 is the cholesterol content in the supernatant.
A0 − A1
Cholesterol binding rate (%) = × 100 (2)
A0
2.5.3 Determination of bile salt binding capacity
Methods refered to Luo et al. [24]. Standard curves were
drawn with cholate content and absorbance. Sodium taurocholate:
y = 0.404x + 0.055, R2 = 0.991; Sodium glycholate: y = 0.624x +
0.045, R2 = 0.995. Among them: c1 is the amount added per μmol;
c2 is residual per μmol.
 Tingting Liu et al. / Journal of Future Foods 2-1 (2022) 61–68 63

T.T. Liu et al. / Journal of Future Foods 2 (2022) 61-68 63

c1 − c2 blood was taken from the eyeballs. Serum TG, TC, LDL-C, HDL-C
Bile salt binding rate (%) = × 100 (3)
c1 and other lipid indexes were detected by kit method.
After the blood was collected, the mice were sacrificed, and their
liver tissues were taken after dissection, washed with normal saline,
2.6 The effect of HQ-HDF in lowering blood lipids in vivo and the water was sucked dry, weighed, and frozen and stored.

2.6.1 Animal feeding

2.6.5 Determination of serum ALT and AST activity in mice
Mice were adaptively feeding at room temperature with a relative
The enzyme activities of ALT and AST in mouse serum were
humidity of 50%−60%. The day and night cycle was 12 h/12 h, during
determined according to the method described in the kit.
which the mice could eat normally.

2.6.6 Determination of SOD, GSH-Px enzyme activity and

2.6.2 Establishment of high-fat model mice
MDA content in serum of mice
One week later, blood was taken from the coccyx to detect the
The mice serum was taken to determine the enzyme activity of
levels of TC and TG in the serum. The mice were randomly divided
SOD, GSH-Px and the content of MDA according to the method
into 6 groups of 7. The mice in the blank group (NG) were fed a described in the kit.
basic feed, and the rest were fed high-fat feed for modeling. The body
weight of the mice was followed regularly, and the modeling time was
2.7 Statistical analysis of data
4 weeks. After 4 weeks, blood was drawn from the coccyx to test the
TC and TG content in the serum to confirm whether a high-fat mouse All experimental data were measured in 3 groups in parallel,
model was established. expressed as mean ± standard deviation, analyzed by SPSS 17.0
software, and used Origin 7.5 to make plots.
2.6.3 Grouping and administration of animals
3. Results and discussion
After the high-fat model was established, the successfully
modeled mice was divided into 5 groups, namely model control 3.1 Basic composition determination
group (MG), positive control group (PG), low (LDG), medium
Table 2 was the basic composition table of HDF and HQ-HDF.
(MDG), and high dietary fiber dose (HDG) group. Each group of
There was no significant difference in the content of TDF in HDF
mice was given intragastric administration once a day. According to and HQ-HDF, but the content of SDF in HQ-HDF was significantly
the Chinese Dietary Nutrition Guide, the average daily intake of 60 increased (P < 0.05). It could be seen that the ultrasound-microwave
kg adults was 30 g, and the gavage volume was 0.2 mL/20 g. NG and assisted enzymatic method changed part of IDF to SDF.
MG mice were given the same volume of ultra-pure water every day.
During this period, the mice could eat and drink freely, and the mice 3.2  Characterization of dietary fiber structure
were weighed and recorded every week.
3.2.1 Scanning electron microscope analysis
Table 1
Grouping situation of mice. Studies have demonstrated that the adsorption activity of DF
Grouping Feed Dosing mainly depends on its internal structure and surface properties [25].
NG basic feed the same volume of ultrapure water Scanning electron microscopes of HDF and HQ-HDF were shown in
MG high-fat feed the same volume of ultrapure water Fig. 1, both types of DF had wrinkles on the surface. HDF mostly had
PG high fat feed 1.0 g/kg simvastatin irregular massive structure, with obvious folds on the surface, but the
LDG high fat feed 0.5 g/kg HQ-HDF structural connection were dense. HQ-HDF had a relatively smaller
MDG high fat feed 1.0 g/kg HQ-HDF
particle size and a looser structure, and its surface structure had obvious
HDG high fat feed 1.5 g/kg HQ-HDF
changes. HQ-HDF exposed the fibrous bundles inside, with more
large cavities, thereby they exposed more polar groups and other water
2.6.4 Determination of serum lipids in mice binding sites to help fibers absorb and retaine water [26]. The results
showed that the ultrasonic-microwave-assisted enzymatic treatment
The mice were fasted the day before the end of the experiment might caused the breakage of glycoside bonds and the degradation of
and had free access to water. They were weighed 24 h later and then fiber macromolecules and maked the structure more loose [27].

Table 2
Analysis of basic components of DF.
Sample Moisture (%) Fat (%) Protein (%) TDF (%) IDF (%) SDF (%)
HDF 4.05 ± 0.11a 1.12 ± 0.06b 4.57 ± 0.09a 88.93 ± 0.73a 84.67 ± 0.45a 4.16 ± 0.10b
HQ-HDF 4.08 ± 0.15a 1.13 ± 0.09b 4.53 ± 0.04a 89.28 ± 0.51a 76.23 ± 0.51c 12.89 ± 0.12a
Note: S Each data was representative of the mean of three replicates. Different letters in the same column represent significant differences (P ＜ 0.05) (the same below).
64 Tingting Liu et al. / Journal of Future Foods 2-1 (2022) 61–68

64 T.T. Liu et al. / Journal of Future Foods 2 (2022) 61-68

a1 a2 a3

× 200 300 µm × 1 000 80 µm × 5 000 10 µm

b1 b2 b3

× 200 300 µm × 1 000 80 µm × 5 000 10 µm

Fig. 1 Scanning electron microscope images of DF. (a) HDF, (b) HQ-HDF.

3.2.2 Particle size analysis Table 3

Particle size distribution of DF.
Particle size is an important aspect of DF. It indirectly Sample D50 (μm) D4,3 (μm) D3,2 (μm) Specific surface area (m2/kg)
affects the physiological functions of the digestive tract, such as HDF 314 331 106 53.94
transportation time, fermentation and fecal excretion [28]. The HQ-HDF 177 223 58.6 97.44
particle size of DF depends on the preparation method of dietary Note: D50 represents the particle size when the cumulative distribution reaches 50%; D4,3:
fiber. Fig. 2 showed in the particle size distribution of HDF and volume average particle size; D3,2: surface area average particle size.

HQ-HDF. Compared with HDF, the peak value of HQ-HDF

particle size distribution moved to the direction of particle size
reduction. The indicated that the particle size decreased, which
might due to the ultrasonic cavitation shearing and the high
energy penetration of microwaves. Physical effects such as
penetration and enzymatic hydrolysis have destroyed part of the
DF with a large particle size and reduced the particle size [29]. In
Table 3, compared with HDF, D 4,3 and D 3,2 of HQ-HDF were
reduced, and the specific surface area was significantly increased.
The indicated that the particle shape of HQ-HDF became irregular
and the texture became Loose, which was the same as the result
of scanning electron microscopy. Studies have shown that
samples with smaller particle size have larger specific surface
area and more exposed groups, which is conducive to improve the
adsorption capacity of samples [30].
10
Differential distribution (%)
3.2.3 SDF relative molecular mass
There was a close relationship between the biological activity
of DF and its molecular weight [31], so the molecular weight of
SDF was determined. The relative molecular mass distributions of
HDF and HQ-HDF were concentrated around 12 172−52 180 and
11 543−49 712 Da, respectively, and their molecular masses were
mainly concentrated around 50 000 Da. However, the proportion
of this component in HQ-HDF declined, and its relative molecular
weight of about 11 543 Da component increased. This might be
caused by the breaking of cellulose and hemicellulose molecular
chains under the action of ultrasound-microwave and enzymes,
which degraded the large molecular in DF into small molecular
components. This leaded to molecular polymerization and a
reduction in relative molecular weight.

HDF
8
HQ-HDF Table 4
6 Relative molecular weight distribution of HDF and HQ-HDF.
4 Sample Peak number Peak time (min) Mw (Da) Relative peak area (%)
1 75.94 52 180 87.36
2
HDF 2 84.25 12 172 6.39
0 3 92.22 22 069 6.24
0.1 1 10 100 1 000
Particle size (µm) 1 76.78 49 712 77.24
HQ-HDF
2 83.87 11 543 22.76
Fig. 2 Particle size distribution of DF.
 Tingting Liu et al. / Journal of Future Foods 2-1 (2022) 61–68 65

T.T. Liu et al. / Journal of Future Foods 2 (2022) 61-68 65

3.3 The physical and chemical functional properties of DF
3.3.1  Pancreatic lipase and α-amylase inhibitory ability
Pancreatic lipase can hydrolyze triglycerides, which can promote
the digestion and absorption of triglycerides in the diet. So inhibiting
its activity is a key way to control hyperlipidemia and obesity. It
could be seen from Table 5 that the ability of HQ-HDF to remove
pancrelipase was significantly higher than that of HDF. HQ-HDF
exposed more binding sites, so pancreatic lipase tended to bind to
DF. In addition, HQ-HDF had a porous structure and a large specific
surface area, which could effectively bind pancrelipase or oil. So
HQ-HDF could inhibit the activity of pancreatic lipase [29].
Table 5
The inhibition ratio of DF on pancreatic lipase activity.
Sample Pancreatic lipase inhibition ability (%)
HDF 52.38 ± 1.67b
HQ-HDF 81.66 ± 2.88a
DF could reduce the cholesterol concentration in the body [33].
Table 7 showed that HDF had a certain binding ability to cholate,
and the ability of HQ-HDF to bind sodium glycocholate and sodium
taurocholate was significantly higher than that of HDF. Compared
with HQ-HDF and HDF, the increase of SDF content could increase
its viscosity, thus reducing the diffusion rate of bile acid and making
it difficult to be absorbed by the body. And the internal structure of
HQ-HDF was more loose and porous, increased the specific surface
area and the binding site of bile salt. So it was easy for bile salt to bind
to it, and then excreted from the body. The bile acid salt combined
with DF inhibited its surface activity in the small intestine, thereby
reduced the formation of lipid micelles. It decreased the ability of
lipid digestion and absorption, which might lead to a decrease in
circulating triglycerides.
3.4 Animal experiment

3.4.1 Establishment of high-fat model mice

3.3.2 Cholesterol adsorption capacity
Fig. 3 showed the changes in serum TC and TG levels in the NG
It is generally accepted that the binding ability of DF with oil and group and model mice before and after modeling. It could be seen
cholesterol is an important factor in evaluating its ability to absorb from the Fig. 3A that the serum levels of TC and TG in the NG group
lipophilic components [32]. The functional groups on the surface mice did not change significantly before and after modeling, and they
of DF may chelate and absorb organic molecules, thereby reducing
were fed high-fat. The blood lipid content in the serum of the mice
liver tissue and plasma cholesterol level [29]. Table 6 showed the
modeled on the feed was significantly increased. It could be seen
cholesterol adsorption capacity of the samples. Cholesterol adsorption
from Fig. 3B that after 4 weeks of molding, the contents of TC and
capacity mainly simulated the stomach environment (pH 2.0) and the
intestinal environment (pH 7.0). It could be seen from the table that TG in serum of mice fed high-fat diet were significantly higher than
cholesterol adsorption capacity of HQ-HDF was significantly higher than those in NG group. The results showed that the high-fat mice were
that of HDF. This might be due to the ultrasound-microwave-assisted successfully modeled.
enzymatic method that making the structure of DF more porous and
4.0 A
porous. DF exposed a large number of groups, the adsorption of 3.5 Before a
After
cholesterol more effective. In addition, the pH value also had a certain 3.0
TC (mmol/L)

effect on cholesterol adsorption capacity of HDF and HQ-HDF. The 2.5 a b

a
2.0
adsorption capacity at pH 7.0 was considerably higher than at pH 2.0. 1.5
It could be deduced that the adsorption capacity of DF for cholesterol 1.0
0.5
was mainly in the intestine.
0.0
Normal Hyperlipidemia
Table 6 Group
The capacity to bind cholesterol of DF in vitro.
Cholesterol binding capacity (mg/g) 8 B
Sample 7 Before b
pH 2.0 pH 7.0 After
6
TG (mmol/L)

HDF 12.75 ± 0.75a 28.89 ± 0.49b 5

HQ-HDF 13.69 ± 0.52a 36.84 ± 0.59a 4
3
2 a a a
3.3.3 Bile salt binding capacity 1
0
Normal Hyperlipidemia
Cholate is generated by the catabolism of cholesterol. The ability Group
of DF to bind to cholic acid speeded up the excretion of cholic acid and Fig. 3 Successful induction of hyperlipidemia in mice. Different letters in the
promoted the decomposition of cholesterol into cholic acid. Thereby same group represent significant differences (P < 0.05).

Table 7
The binding capacity of cholate by DF.
Cholate binding rate (%) Cholate binding capacity (mg/g)
Sample
Sodium glycocholate Sodium taurocholate Sodium glycocholate Sodium taurocholate
HDF 36.06 ± 0.85b 37.19 ± 0.71b 15.01 ± 0.23b 22.58 ± 0.36b
HQ-HDF 41.50 ± 1.54a 44.54 ± 0.44a 17.11 ± 0.14a 27.33 ± 0.25a
66 Tingting Liu et al. / Journal of Future Foods 2-1 (2022) 61–68

66 T.T. Liu et al. / Journal of Future Foods 2 (2022) 61-68

3.4.2 Effects of HQ-HDF on serum lipids in mice Table 9

Effect of HQ-HDF on ALT and AST activities in mice.
Elevated levels of TC and TG are the main factors leading to Grouping ALT (U/L) AST (U/L)
hyperlipidemia. Excess LDL-C tends to accumulate and oxidize in NG 13.71 ± 0.32e 26.49 ± 1.72e

blood vessels, and may lead to atherosclerosis. HDL-C could transport MG 45.79 ± 1.03a 81.06 ± 1.97a
c
cholesterol from tissues and blood vessels to the liver, where it is PG 23.39 ± 1.55 55.62 ± 1.74c
b
LDG 32.97 ± 1.16 68.39 ± 1.41b
metabolized and broken down bile acids that facilitate its excretion. It
MDG 25.05 ± 2.01c 57.72 ± 1.03c
was a lipoprotein that can resist atherosclerosis [34]. It could be seen
HDG 20.89 ± 1.49d 51.98 ± 1.88d
from Table 8 that compared with the NG group, the concentrations
of TC, TG, and LDL-C in the serum of MG mice were significantly
increased (P < 0.01), and the content of HDL-C was significantly 3.4.4 Effects of HQ-HDF on serum antioxidant index of mice
reduced by 43.44% (P < 0.01). It showed that the high-fat diet
caused hyperlipidemia in the model group. Compared with the MG A high-fat diet can cause oxidative stress response in tissues,
group, HQ-HDF could significantly reduce blood lipid TC, TG leading to hyperlipidemia and cardiovascular diseases [38]. Many
and LDL-C levels (P < 0.01), and significantly increase HDL-C studies have shown that antioxidant enzymes (such as SOD and
levels (P < 0.05). There was no significant differences between GSH-Px) in blood contribute to improving antioxidant defense
the positive control group of Vastatin (P > 0.05), and both could mechanisms [39]. There is considerable evidence that the activity
effectively improve the blood lipid level. The results showed that of antioxidant enzymes is lower in animals on a high-fat diet. In
HQ-HDF could improve the lipid accumulation of mice serum addition, MDA is a lipid peroxide and is considered to be a marker of
induced by high-fat diet to a certain extent.
oxidative stress and liver injury [38].
Antifcial intelligence (AI) is usually used in medicine to assess the
As shown in Table 10, the activities of SOD and GSH-Px in
degree of atherosclerosis [35]. The larger the AI value, the higher the
serum of mice in MG group were significantly lower than those
degree of atherosclerosis. It could be seen from Table 8 that the AI value of
the MG group was considerably higher than that of the NG group (P < 0.01), in NG group, and the content of MDA was significantly increased
indicating that the high-fat diet could cause severe atherosclerosis in mice. (P < 0.01). The results showed that high-fat diet increased oxidative
Compared with the MG group, the AI values of mice in the LDG, MDG, stress response. Compared with MG group, the MDA content in HDG
and HDG groups were significantly lower (P < 0.01), indicating that HQ- group was significantly decreased (P < 0.01), and the ativities of
HDF could improve and decrease the degree of atherosclerosis. SOD and GSH-Px in medium and high dose groups were increased
more obviously, and their antioxidant activities were increased in a
3.4.3 Effects of HQ-HDF on the activities of serum ALT and concentration-dependent manner. The activities of SOD and GSH-Px
AST in mice and MDA content in HDG group were close to those in simvastatin
PG group (P > 0.05). The results showed that HQ-HDF with high dose
The determination of ALT and AST activity can reflect liver damage had stronger antioxidant activity. HQ-HDF significantly counteracted
in mice [36,37]. The toxic effect of a high-fat diet on the liver is related oxidative stress by reducing the level of lipid peroxidation products
to the permeability of the liver cell membrane. The rupture of the cell
and promoting the activity of both enzymatic and non-enzymatic
membrane leads to the dissolution of intracellular enzymes into the
antioxidants.
blood [35], thereby increasing the enzyme activity in the blood.
Table 9 showed that compared with the NG group mice, the
Table 10
ALT and AST activities in the serum of the MG group mice were Effect of HQ-HDF on SOD and MDA levels in mice.
significantly increased (P < 0.01). It indicated that the high-fat diet Grouping SOD (U/mL) GSH-Px (U/mL) MDA (nmol/mL)
could cause severe oxidative damage in the mice, liver damage and
NG 24.71 ± 2.48a 368.97 ± 10.16a 7.26 ± 0.76d
impaired function. Compared with the MG group, the PG group and
MG 10.24 ± 0.77d 295.70 ± 17.98c 14.32 ± 0.94a
HQ-HDF each dose group could reduce the activity of ALT and AST ab b
PG 21.13 ± 0.81 330.87 ± 9.28 8.84 ± 0.27c
to a certain extent (P < 0.01). Both simvastatin and HQ-HDF could d c
LDG 11.10 ± 1.16 299.40 ± 12.23 10.45 ± 0.77b
maintain the integrity of mouse cell membrane and restrict the release c bc
MDG 17.55 ± 1.76 313.11 ± 4.93 9.85 ± 0.80bc
of transaminase. HQ-HDF in the high-dose group could effectively b b
HDG 21.62 ± 0.74 335.60 ± 16.45 8.55 ± 0.46c
prevent hyperlipidemia and liver damage.

Table 8
Effect of HQ-HDF on serum lipid levels in mice.
Grouping TC (mmol/L) TG (mmol/L) HDL-C (mmol/L) LDL-C (mmol/L) AI
NG 2.68 ± 0.54e 1.74 ± 0.25b 2.67 ± 0.35a 0.95 ± 0.20d 0.03 ± 0.00e
MG 6.67 ± 0.99a 7.03 ± 0.24a 1.51 ± 0.42c 1.85 ± 0.26a 2.71 ± 0.20a
d c ab c
PG 3.80 ± 0.47 1.32 ± 0.26 2.32 ± 0.23 1.09 ± 0.14 0.69 ± 0.07d
b b c b
LDG 4.99 ± 0.37 1.88 ± 0.57 1.76 ± 0.38 1.37 ± 0.14 1.73 ± 0.09b
cd bc b bc
MDG 4.14 ± 0.13 1.52 ± 0.11 2.21 ± 0.54 1.23 ± 0.25 1.26 ± 0.06c
HDG 4.47 ± 0.27bc 1.53 ± 0.31bc 1.95 ± 0.24bc 1.34 ± 0.19b 1.36 ± 0.31c
 Tingting Liu et al. / Journal of Future Foods 2-1 (2022) 61–68 67
T.T. Liu et al. / Journal of Future Foods 2 (2022) 61-68
A B
D E
Fig. 4 Effect of HQ-HDF on hepatic tissue morphology in mice. (A) NG, (B) MG, (C) PG, (D) LDG, (E) MDG, (F) HDG.
3.4.5 The effect of HQ-HDF on liver histopathology in mice
The pathological observation of mice liver tissue (magnification
× 100), the result was presented in Fig. 4. The livers of mice in the
NG group did not show obvious pathological abnormalities, and were
normal liver tissue structures [38]. Sections of mice in the MG group
showed steatosis, showing a large number of fatty white particles,
swollen liver cells, and larger in size than the NG group. In the PG
group, the liver cell structure was intact, the white fat particles in
the cytoplasm were significantly reduced, and the liver cell staining
became darker. The liver of the mice in the LDG group improved
slightly. Pathological features existed but the white fat particles in
the cytoplasm were reduced. The degree of degeneration of liver cells
in the MDG group mice was reduced, and the fat particles were also
significantly reduced. The liver of the HDG group mice had a small
amount of cell degeneration and fewer white fat particles. HQ-HDF
could improve the fatty degeneration of hepatocytes in hyperlipidemia
mice. With the increase of the dose of HQ-HDF, its effect became more
obvious, and the improvement effect was best in the high dose group.
4. Conclusions
Ultrasound-microwave assisted enzymatic method could
improve the pancreatic lipase inhibition capacity, cholesterol and
bile acid adsorption capacity of HDF. HQ-HDF had a better in vitro
blood lipid-lowering effect than HDF, which might be linked to
its porous structure and crystallinity. The decrease resulted in an
increase in dietary fiber binding sites. The levels of TC (P < 0.01),
TG (P < 0.01), LDL-C (P < 0.01, P < 0.05) and HDL-C in serum
of hyperlipidemia mice were significantly increased in medium and
high dose groups (P < 0.01). The above results showed that HQ-HDF
could improve serum lipid metabolism. The high dose HQ-
HDF group could significantly reduce the activities of ALT and
AST (P < 0.01), significantly increased the activities of SOD and
GSH-Px (P < 0.01), and significantly reduced the content of lipid
67
C
F
peroxide MDA (P < 0.01). The results showed that HQ-HDF
had protective effect on hepatocytes. HQ-HDF had a significant
improvement effect on oxidative stress and liver damage caused by
hyperlipidemia, so that it could achieve a better effect of lowering
blood lipids.
Declaration of competing interest
The authors declare that they have no conflflict of interest.
Acknowledgments
The study was financially supported by the Key Projects of the
National Research and Development Program of China (Grant No.
2018YFD0400204).
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