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Citation: Wang, H.; Li, H.; Li, Z.;
Feng, L.; Peng, L. Evaluation of
Prebiotic Activity of Stellariae Radix
Polysaccharides and Its Effects on
Gut Microbiota. Nutrients 2023,15,
4843. https://doi.org/10.3390/
nu15224843
Academic Editor: Stefano
Guandalini
Received: 28 September 2023
Revised: 30 October 2023
Accepted: 14 November 2023
Published: 20 November 2023
Copyright: © 2023 by the authors.
Licensee MDPI, Basel, Switzerland.
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Attribution (CC BY) license (https://
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4.0/).
nutrients
Article
Evaluation of Prebiotic Activity of Stellariae Radix
Polysaccharides and Its Effects on Gut Microbiota
Hong Wang 1,2, Haishan Li 1, Zhenkai Li 1, Lu Feng 1and Li Peng 1,3,*
1School of Life Sciences, Ningxia University, Yinchuan 750021, China; wh12100912@163.com (H.W.);
lhs985233@163.com (H.L.)
2College of Resource and Environment and Life Science, Ningxia Normal University, Guyuan 756000, China
3Ningxia Natural Medicine Engineering Technology Research Center, Yinchuan 750021, China
*Correspondence: pengli1124@163.com
Abstract:
This study aims to evaluate the prebiotic potential of polysaccharides derived from Stellar-
iae Radix (SRPs) and explore their influence on the gut microbiota composition in mice. Lactobacillus
acidophilus and Bifidobacterium longum were cultivated in an MRS medium, while their growth kinet-
ics, clumping behavior, sugar utilization, pH variation, growth density, and probiotic index were
meticulously monitored. Additionally, the impact of crude Stellariae Radix polysaccharides (CSRP)
on the richness and diversity of gut microbiota in mice was assessed via 16S rDNA sequencing. The
results demonstrated the remarkable ability of CSRPs to stimulate the proliferation of Lactobacillus
acidophilus and Bifidobacterium longum. Moreover, the oral administration of CSRPs to mice led to
a noticeable increase in beneficial bacterial populations and a concurrent decrease in detrimental
bacterial populations within the intestinal flora. These findings provided an initial validation of
CSRPs as a promising agent in maintaining the equilibrium of gut microbiota in mice, thereby offer-
ing a substantial theoretical foundation for developing Stellariae Radix as a prebiotic ingredient in
various applications, including food, healthcare products, and animal feed. Furthermore, this study
presented novel insights for the exploration and utilization of Stellariae Radix resources.
Keywords: Stellariae Radix; polysaccharides; prebiotic activity; gut microbiota
1. Introduction
Lactobacillus acidophilus and Bifidobacterium longum are representative probiotics found
in the intestines, and they offer numerous benefits for human health [
1
]. Probiotics pro-
duce beneficial substances for the intestines, such as short-chain fatty acids (SCFAs), lactic
acid, and other acidic metabolites, through prebiotics. These substances play a crucial
role in maintaining intestinal health [
2
]. Prebiotics are food components that selectively
ferment and alter the composition and activity of beneficial bacteria in the intestines. They
mainly consist of non-digestible oligosaccharides and oligose [
3
]. Many studies have
demonstrated that regulating the gut microbiota can reduce the risk of chronic metabolic
diseases, including type 2 diabetes, obesity, and non-alcoholic fatty liver disease [
4
]. The
gut microbiota prevents the infection of external pathogens through direct mechanisms
(such as competition for nutrients and niches) and indirect mechanisms (such as enhancing
the host’s defense) [
5
]. Therefore, maintaining and regulating the balance of the intesti-
nal microecology is of great significance in disease prevention and promoting overall
host health.
Polysaccharides are complex carbohydrates with various biological activities [
6
]. Plant
polysaccharides are typically not directly or fully absorbed by the intestine. Instead, they
are broken down into SCFAs and other beneficial metabolites through interactions with
intestinal microbes and are then absorbed by the intestine [
7
]. In recent years, numerous
studies have suggested that non-digestible polysaccharides may serve as potential prebi-
otics for the gut microbiota, leading to changes in microbial structure and composition [
8
].
Nutrients 2023,15, 4843. https://doi.org/10.3390/nu15224843 https://www.mdpi.com/journal/nutrients
Nutrients 2023,15, 4843 2 of 12
For instance, Lycium barbarum polysaccharides have been found to promote the growth
of Lactobacillus acidophilus and increase the proportion of intestinal probiotics, such as
Akkermansia,Lactobacillus acidophilus, and Prevotellaceae, in mice [
9
]. Similarly, Dendrobium
huoshanense polysaccharides have been shown to stimulate the production of cytokines,
induce the proliferation and differentiation of immune cells in the small intestine, increase
the relative abundance of Lactobacillus,Prevotella, and Porphyromonas in the mouse colon,
and reduce the abundance of Helicobacter and Chlorotriazole [
10
]. Therefore, further research
is needed to understand the role of polysaccharides in the gut microbiota and their impact
on overall body health.
The traditional Chinese medicine Stellariae Radix is derived from the dried root of Stel-
laria dichotoma L. var. lanceolata Bge. [
11
]. It is characterized by its cold and sweet properties
and is known for its efficacy in treating infantile malnutrition and bone tuberculosis [
12
].
With the advancement of modern medical research, the anti-inflammatory, anti-allergic,
and anti-cancer effects of Stellariae Radix have been continuously discovered, enhancing
its medicinal value. The effective components and pharmacological effects of Stellariae
Radix, as a traditional Chinese medicine, have been extensively reported [
13
–
15
]. However,
there has been no research conducted on the impact of Stellariae Radix polysaccharides
(SRPs) on the intestinal microflora.
In this study, Lactobacillus acidophilus and Bifidobacterium longum were cultured in an
MRS medium to monitor the dynamic changes in the growth curve, clump count, pH,
growth density, and probiotic index of these two probiotics. Additionally, the effects of
crude Stellariae Radix polysaccharides (CSRPs) on the abundance and diversity of gut
microbiota in mice were analyzed using 16S rDNA. These findings provide a theoretical
basis for the diversified development of food, health products, feed, and other related fields.
Furthermore, they offer new insights for the development and utilization of Stellariae
Radix resources.
2. Materials and Methods
2.1. Materials
Stellariae Radix was purchased from Ningxia, China. Lactobacillus acidophilus
(CGMCC1.3013) and Bifidobacterium longum (CGMCC1.1878) were purchased from the
China General Microbiological Culture Collection Center. The bacterial strains were inocu-
lated in a broth (MRS) medium and activated under anaerobic conditions (37 ◦C, 12 h).
2.2. Preparation Process of CSRPs
Stellariae Radix medicinal materials were crushed through a 40-mesh sieve and de-
greased by supercritical CO
2
extraction (a pressure of 35 MPa, an extraction temperature
of 47
◦
C, a CO
2
flow rate of 50 L/h, and extraction for 3 h). Then, ultrasonic-assisted
extraction was used after concentration and fractional alcohol precipitation, and CSRPs
were obtained by the Sevag deproteinization method [16].
2.3. Evaluation of Prebiotic Activity In Vitro
The growth and acid production capacity of Lactobacillus acidophilus and Bifidobacterium
longum were determined to evaluate the prebiotic activity of CSRPs [
17
]. MRS carbohydrate-
free broth (Qingdao Top Biotech Co., Ltd., Qingdao, China) was used as the basal medium.
In order to cultivate Bifidobacterium longum, 5 g/L cysteine was added to the MRS medium
as the reducing agent. The existing literature shows that the two probiotics have an optimal
growth concentration of 2.5% (w/v) inulin; thus, 2.5% (w/v) Glc (negative control without
inulin) and 2.5% (w/v) inulin are selected as the control groups [
9
]. Then, 2.5% (w/v)
inulin (MRS-L) and 2.5% (w/v) Glc with different concentrations of CSRPs (2.5%, 5%,
10%, and 15% (w/v)) were added to the MRS broth medium as carbohydrate sources, and
MRS-L was used as the control group. The broth medium was inoculated with 5% (v/v)
(
1−2×109CFU/mL
) activated Lactobacillus acidophilus or Bifidobacterium longum. And
the culture medium was incubated under anaerobic conditions for 48 h (37
◦
C, 120 rpm),
Nutrients 2023,15, 4843 3 of 12
and random sampling was performed every 3 h to determine the OD value of the culture
medium. In addition, the logistic model in Origin Pro 2019b software [
18
] was used, and
the iterative algorithm used was the Levernberg–Marquardt optimization algorithm. And
the algorithm (expression (1)) was as follows, and the fitting growth curves of two bacteria
were obtained.
y=A2+ (A1−A2)/(1 + (X/X0)ˆp) (1)
In the formula, A
1
, A
2
, X
0
, and p are parameters; A
1
is the deviation degree between
the real curve and the model; A
2
is the growth maximum predicted by the model; X
0
is
the inflection point time, representing the maximum growth rate at this time; and p is the
growth rate coefficient, and the curve has a maximum slope at the crossing point (X0, A2),
indicating the maximum growth rate.
The growth number of two probiotic strains was determined via the plate counting
method, expressed as log CFU/mL. The control groups are 2.5% (w/v) Glc and 2.5% (w/v)
inulin. The broths were inoculated with 5% (v/v) (10
×
10
9
CFU/mL) of stationary-phase
Lactobacillus acidophilus and Bifidobacterium longum. The cultures were then incubated at
37 ◦C
for 24 h with agitation (120 rpm) under anaerobic conditions. After incubation, serial
dilutions (10
−1
–10
−8
) were performed using sterile normal saline and plated on an MRS
agar at 37
◦
C for 48 h under anaerobic conditions. The results were recorded as CFU/mL
of culture. The acid production ability and growth of the two probiotic strains with pH and
OD values were determined, and the probiotic values (PIs) were calculated.
2.4. Evaluation of Prebiotic Activity In Vivo
Clean-grade ICR male mice (8 weeks old; weight: 20.0
±
2.0 g) were purchased
from the Experimental Animal Center of Ningxia Medical University (Ningxia, China)
(Certificate No. SCXK Ningxia 2020-0001). The animal housing and feeding conditions
were as follows: temperature of 20–26
◦
C, relative humidity of 40–70%, alternating light and
dark for 12 h, free drinking water, and feeding with standard blocontrol
maintenance feed.
The mice were randomly divided into four groups (6 in each group), respectively, the
control group (CK), the low-dose group (L), the middle-dose group (M), and the high-dose
group (H), and were allowed to acclimate for 1 week before the experiment. The control
group (intragastric administration of normal saline), L group (CSRPs: 1 g/kg), M group
(CSRPs: 2 g/kg), and H group (CSRPs: 4 g/kg), underwent an intragastric administration
of 0.1 mL/10 g continuously for 14 days. After 24 h from the last administration, the blood
was taken from the eyeballs of the mice, and fresh feces were collected.
2.5. Composition and Diversity of Gut Microbiota
The 16S rDNA amplicon sequencing technology was used to study microbial com-
position and structure in the mice’s intestine. Total microbial genomic DNA was ex-
tracted from the samples using the E.Z.N.A.
®
soil DNA kit (Omega Bio-tek, Norcross,
GA, USA) according to the manufacturer’s instructions. The quality and concentration
of DNA were determined with 1.0% agarose gel electropHoresis and a NanoDrop
®
ND-
2000 spectropHotometer (Thermo Scientific Inc., Waltham, MA, USA). The hypervariable
region, V3-V4, of the bacterial 16S rRNA gene was amplified with the primer pairs 338F
(
50-ACTCCTACGGGAGGCAGCAG-30
) and 806R (5
0
-GGACTACHVGGGTWTCTAAT-3
0
)
by an ABI GeneAmp
®
9700 PCR thermocycler (ABI, Los Angeles, CA, USA). All samples
were amplified in triplicate after mixing PCR products of the same sample, and the PCR
product was extracted from a 2% agarose gel and purified using the AxyPrep DNA Gel
Extraction Kit (Axygen Biosciences, Union City, CA, USA) according to the manufacturer’s
instructions and quantified using a Quantus
™
Fluorometer (Promega, Madison, WI, USA).
An Illumina Pair-End library was constructed, and the sub-library was expanded to perform
pairing. Online measurements on the Illumina MiSeq platform were carried out.
Using UPARSE 7.1, the sequences were clustered into operational taxonomic units
(OTUs) with 97% consistency. Bacterial annotation of OTU sequences was carried out, and
a bacterial annotation analysis was carried out using the Mothur method and the SSUrRNA
Nutrients 2023,15, 4843 4 of 12
database of SILVA132 to obtain taxonomic information and the composition of the bacterial
colonies of the samples to be tested. Alpha diversity indices for evaluating gut microbial
community richness were calculated. A beta diversity analysis was used to investigate the
structural variations in the microbial communities using the UniFrac distance.
2.6. Statistical Analysis
Data are presented as the mean
±
SD, and the statistical analysis was performed
using Origin 2019 software (OriginLab Corporation, Northampton, MA, USA). A one-way
analysis of variance (ANOVA) plus the post hoc Duncan’s test (SPSS software,
version 22.0
)
were used to evaluate the statistical significance. The Wilcox rank sum test and Tukey
test were used to analyze the differences in bacterial diversity between the groups. A
bioinformatic analysis of the gut microbiota was carried out using the Majorbio Cloud
platform (https://cloud.majorbio.com, accessed on 30 September 2021).
3. Results
3.1. Composition of Polysaccharide from Stellariae Radix
The SRP consists of Gal, Glc, Xyl, Fru, Man, and Rha, with molar percentages of
61.86%, 32.51%, 4.77%, 0.39%, 0.28%, and 0.19%, respectively. It has a molecular weight of
31,309 Da, and the sugar ring structure of pyran sugar contains
α
-configuration glycosidic
bonds and β-configuration glycosidic bonds [19].
3.2. Evaluation of Prebiotic Activity In Vitro
3.2.1. Fitting of Growth Curve
The growth curves of the two probiotic strains are plotted in Figure 1, and the model
fitting had determination coefficient (R
2
) values greater than 0.9, indicating a good fitting
effect. Figure 1a shows the growth curve of Lactobacillus acidophilus with different carbon
(2.5% (w/v) inulin (MRS-L), 2.5% (w/v) Glc, and different concentrations of CSRP (2.5%, 5%,
10%, and 15% (w/v))) sources. A lag phase occurred from 0 to 3 h, followed by a log phase
from 3 to 12 h. After 12 h, the growth rate slowed down, and from 15 to 48 h, a stationary
phase was observed. Figure 1b depicts the growth curve of Bifidobacterium longum with
different carbon (2.5% (w/v) inulin (MRS-L), 2.5% (w/v) Glc, and different concentrations
of CSRP (2.5%, 5%, 10%, and 15% (w/v))) sources. In general, an extended lag phase was
observed from 3 to 9 h, followed by a log period from 9 to 18 h. After 18 h, the growth rate
decreased, and from 21 to 48 h, a stationary phase was observed.
Nutrients 2023, 15, x FOR PEER REVIEW 5 of 13
the stable phase. These results indicate that the CSRP groups, at each concentration, eec-
tively enhance the metabolic rate of the two benecial strains and promote their prolifer-
ation compared to the control group (2.5% inulin and 2.5% Glc).
(a)
(b)
Figure 1. (a) Growth ing curves of Lactobacillus acidophilus; (b) growth ing curves of Bidobac-
terium longum. (The concentration 10% CSRPs is the optimal concentration for Lactobacillus acidophi-
lus and Bidobacterium longum growth).
Table 1. Nonlinear ing parameters of growth kinetics equation.
.
Parameters
A1
A2
X0
p
R2
Lactobacillus aci-
dophilus
2.5% inulin
0.024 ± 0.06
0.727 ± 0.02
4.892 ± 0.55
3.951 ± 1.23
0.967
2.5% Glc
0.024 ± 0.06
1.140 ± 0.03
5.668 ± 0.46
2.857 ± 0.54
0.985
2.5% CSRP
0.021 ± 0.03
0.954 ± 0.01
4.307 ± 0.16
4.035 ± 0.37
0.996
5% CSRP
0.032 ± 0.04
1.391 ± 0.02
5.244 ± 0.20
4.138 ± 0.54
0.996
10% CSRP
0.008 ± 0.02
1.872 ± 0.02
7.345 ± 0.12
3.792 ± 0.20
0.999
15% CSRP
0.009 ± 0.02
1.801 ± 0.02
8.495 ± 0.17
3.930 ± 0.27
0.998
Bifidobacterium
longum
2.5% inulin
0.044 ± 0.04
1.923 ± 0.13
25.043 ± 1.34
4.058 ± 0.66
0.991
2.5% Glc
0.027 ± 0.04
1.487 ± 0.04
20.010 ± 0.42
4.181 ± 0.36
0.997
2.5% CSRP
0.005 ± 0.04
2.192 ± 0.05
14.899 ± 0.31
6.441 ± 0.74
0.995
5% CSRP
0.020 ± 0.03
2.217 ± 0.02
11.330 ± 0.14
7.986 ± 0.67
0.998
10% CSRP
0.002 ± 0.05
2.360 ± 0.05
11.820 ± 0.26
6.017 ± 0.69
0.996
15% CSRP
0.016 ± 0.03
1.711 ± 0.05
17.966 ± 0.35
7.601 ± 0.96
0.990
Note: parameters’ meanings were shown in experimental methods 2.3.
3.2.2. Eects of CSRPs on Growth of Lactobacillus Acidophilus and Bidobacterium
Longum
The eects of CSRPs on the growth of Lactobacillus acidophilus and Bidobacterium
longum were evaluated by measuring the total number of viable bacteria (log10CFU/mL).
According to Table 2, the maximum growth of Lactobacillus acidophilus and Bidobacterium
longum was observed at a concentration of 10% CSRPs, while the growth showed a down-
ward trend at the highest concentration.
Figure 1.
(
a
) Growth fitting curves of Lactobacillus acidophilus; (
b
) growth fitting curves of Bifidobac-
terium longum. (The concentration 10% CSRPs is the optimal concentration for Lactobacillus acidophilus
and Bifidobacterium longum growth).
Nutrients 2023,15, 4843 5 of 12
By referring to Table 1, it can be seen that both probiotic strains exhibited rapid growth
in the medium with different carbon (2.5% (w/v) inulin (MRS-L), 2.5% (w/v) Glc, and
different concentrations of CSRP (2.5%, 5%, 10%, and 15% (w/v))) sources. In the medium
with different carbon sources for Lactobacillus acidophilus, the addition of the 2.5% and
5% CSRP
groups resulted in a faster entry into the logarithmic growth phase compared to
the control group (2.5% inulin and 2.5% Glc) (CSRP group X
0
< control group X
0
). However,
the addition of the 10% and 15% CSRP groups entered the log phase later than the control
group, and the former also entered the stationary phase earlier. Similarly, Bifidobacterium
longum in different carbon-source mediums with different concentrations of CSRP groups
entered the log phase faster than the control group (2.5% inulin and 2.5% Glc) (CSRP group
X
0
< control group X
0
). Furthermore, the maximum growth number of both probiotic
strains in each CSRP concentration group was greater than the control group (the CSRP
group’s p-value was greater than that of the control group) after entering the stable phase.
These results indicate that the CSRP groups, at each concentration, effectively enhance the
metabolic rate of the two beneficial strains and promote their proliferation compared to the
control group (2.5% inulin and 2.5% Glc).
Table 1. Nonlinear fitting parameters of growth kinetics equation.
. Parameters A1A2X0pR2
Lactobacillus acidophilus
2.5% inulin 0.024 ±0.06 0.727 ±0.02 4.892 ±0.55 3.951 ±1.23 0.967
2.5% Glc 0.024 ±0.06 1.140 ±0.03 5.668 ±0.46 2.857 ±0.54 0.985
2.5% CSRP 0.021 ±0.03 0.954 ±0.01 4.307 ±0.16 4.035 ±0.37 0.996
5% CSRP 0.032 ±0.04 1.391 ±0.02 5.244 ±0.20 4.138 ±0.54 0.996
10% CSRP 0.008 ±0.02 1.872 ±0.02 7.345 ±0.12 3.792 ±0.20 0.999
15% CSRP 0.009 ±0.02 1.801 ±0.02 8.495 ±0.17 3.930 ±0.27 0.998
Bifidobacterium longum
2.5% inulin 0.044 ±0.04 1.923 ±0.13 25.043 ±1.34 4.058 ±0.66 0.991
2.5% Glc 0.027 ±0.04 1.487 ±0.04 20.010 ±0.42 4.181 ±0.36 0.997
2.5% CSRP 0.005 ±0.04 2.192 ±0.05 14.899 ±0.31 6.441 ±0.74 0.995
5% CSRP 0.020 ±0.03 2.217 ±0.02 11.330 ±0.14 7.986 ±0.67 0.998
10% CSRP 0.002 ±0.05 2.360 ±0.05 11.820 ±0.26 6.017 ±0.69 0.996
15% CSRP 0.016 ±0.03 1.711 ±0.05 17.966 ±0.35 7.601 ±0.96 0.990
Note: parameters’ meanings were shown in experimental methods 2.3.
3.2.2. Effects of CSRPs on Growth of Lactobacillus Acidophilus and
Bifidobacterium Longum
The effects of CSRPs on the growth of Lactobacillus acidophilus and Bifidobacterium
longum were evaluated by measuring the total number of viable bacteria (log10 CFU/mL).
According to Table 2, the maximum growth of Lactobacillus acidophilus and Bifidobacterium
longum was observed at a concentration of 10% CSRPs, while the growth showed a down-
ward trend at the highest concentration.
Table 2. Effects of CSRPs on the growth of Lactobacillus acidophilus and Bifidobacterium longum.
Bacterial Strain
Control Group (%) Concentration of CSRP (%)
2.5% Inulin 2.5% Glc 2.5% 5% 10% 15%
Lactobacillus acidophilus 1.36 ±0.10 b 0.95 ±0.05 0.8 ±0.17 a 1.76 ±0.02 c 2.39 ±0.03 e 2.06 ±0.02 d
Bifidobacterium longum 1.49 ±0.20 a 0.84 ±0.03 2.08 ±0.03 b 2.36 ±0. 03 d 2.44 ±0.02 e 2.27 ±0.03 c
Note: values are expressed as mean values of log10
±
SD (CFU) per milliliter of MRS. And because a, b, c, d, and
e are in the same row, the average values of different letters were significantly different when p< 0.05, n= 3.
Nutrients 2023,15, 4843 6 of 12
3.2.3. Effects of CSRPs on Acid Production of Two Probiotics
Polysaccharides serve as a carbon source that can be decomposed and utilized by
probiotics. During this process, the probiotics produce short-chain fatty acids and other
substances, leading to a decrease in pH. To evaluate the growth of probiotics, the changes in
pH were monitored. In Figure 2, the pH changes of Lactobacillus acidophilus (Figure 2a) and
Bifidobacterium longum (Figure 2b) are shown when CSRPs were used as a carbon source,
replacing glucose in the MRS medium.
Nutrients 2023, 15, x FOR PEER REVIEW 6 of 13
Table 2. Eects of CSRPs on the growth of Lactobacillus acidophilus and Bidobacterium longum.
Bacterial Strain
Control Group (%)
Concentration of CSRP (%)
2.5% Inulin
2.5% Glc
2.5%
5%
10%
15%
Lactobacillus acidophilus
1.36 ± 0.10 b
0.95 ± 0.05
0.8 ± 0.17 a
1.76 ± 0.02 c
2.39 ± 0.03 e
2.06 ± 0.02 d
Bifidobacterium longum
1.49 ± 0.20 a
0.84 ± 0.03
2.08 ± 0.03 b
2.36 ± 0. 03 d
2.44 ± 0.02 e
2.27 ± 0.03 c
Note: values are expressed as mean values of log10 ± SD (CFU) per milliliter of MRS. And because
a, b, c, d, and e are in the same row, the average values of dierent leers were signicantly dierent
when p < 0.05, n = 3.
3.2.3. Eects of CSRPs on Acid Production of Two Probiotics
Polysaccharides serve as a carbon source that can be decomposed and utilized by
probiotics. During this process, the probiotics produce short-chain fay acids and other
substances, leading to a decrease in pH. To evaluate the growth of probiotics, the changes
in pH were monitored. In Figure 2, the pH changes of Lactobacillus acidophilus (Figure 2a)
and Bidobacterium longum (Figure 2b) are shown when CSRPs were used as a carbon
source, replacing glucose in the MRS medium.
For Lactobacillus acidophilus, the pH of the 10% CSRP group was signicantly lower
than that of the control group (2.5% inulin and 2.5% Glc). Similarly, for Bidobacterium
longum, the pH of the dierent concentrations of CSRP groups was signicantly lower
than that of the control group (2.5% inulin and 2.5% Glc). These results indicate that CSRPs
can promote the growth of probiotics by reducing the pH of the medium and consuming
the carbon source.
(a)
(b)
Figure 2. (a) Eects of dierent concentrations of CSRPs on acid production of Lactobacillus acidoph-
ilus; (b) eects of dierent concentrations of CSRPs on acid production of Bidobacterium longum. (*
p < 0.05, ** p < 0.01*** p < 0.001). (10% CSRPs can promote acid production by probiotics.).
3.2.4. Eects of Dierent Concentrations of CSRPs on Growth Density of two Probiotic
Strains
The growth of probiotics can also be assessed by measuring the optical density (OD)
values of the culture medium, as they reect the growth rate of the strains. In Figure 3a,b,
the OD values of the culture medium for both Lactobacillus acidophilus and Bidobacterium
longum are recorded. It was observed that with the increase in CSRP concentrations, the
OD values gradually increased, reaching their maximum value in the 10% CSRP group.
This indicates that higher concentrations of CSRPs promote the growth of probiotics, re-
sulting in higher OD values in the culture medium.
Figure 2.
(
a
) Effects of different concentrations of CSRPs on acid production of Lactobacillus acidophilus;
(
b
) effects of different concentrations of CSRPs on acid production of Bifidobacterium longum. (
*p< 0.05,
** p< 0.01, *** p< 0.001). (10% CSRPs can promote acid production by probiotics).
For Lactobacillus acidophilus, the pH of the 10% CSRP group was significantly lower
than that of the control group (2.5% inulin and 2.5% Glc). Similarly, for Bifidobacterium
longum, the pH of the different concentrations of CSRP groups was significantly lower than
that of the control group (2.5% inulin and 2.5% Glc). These results indicate that CSRPs can
promote the growth of probiotics by reducing the pH of the medium and consuming the
carbon source.
3.2.4. Effects of Different Concentrations of CSRPs on Growth Density of two
Probiotic Strains
The growth of probiotics can also be assessed by measuring the optical density (OD)
values of the culture medium, as they reflect the growth rate of the strains. In Figure 3a,b,
the OD values of the culture medium for both Lactobacillus acidophilus and Bifidobacterium
longum are recorded. It was observed that with the increase in CSRP concentrations, the OD
values gradually increased, reaching their maximum value in the 10% CSRP group. This
indicates that higher concentrations of CSRPs promote the growth of probiotics, resulting
in higher OD values in the culture medium.
3.2.5. Probiotic Index Calculation of Different Concentrations of CSRPs on Two Probiotic
Strains
As shown in the Table 3, the results of the two probiotic strains showed that the
addition of 10% CSRPs to the medium had the best effect on promoting the probiotic
strains’ growth.
Nutrients 2023,15, 4843 7 of 12
Nutrients 2023, 15, x FOR PEER REVIEW 7 of 13
(a)
(b)
Figure 3. (a) Eects of dierent concentrations of CSRPs on growth density of Lactobacillus acidophi-
lus; (b) eects of dierent concentrations of CSRPs on growth density of Bidobacterium longum. ((*
p < 0.05, ** p < 0.01, *** p < 0.001). (10% CSRP can promote the growth of probiotics).
3.2.5. Probiotic Index Calculation of Dierent Concentrations of CSRPs on Two Probiotic
Strains
As shown in the Table 3, the results of the two probiotic strains showed that the ad-
dition of 10% CSRPs to the medium had the best eect on promoting the probiotic strains’
growth.
Table 3. Eects of CSRPs on PI values of probiotics.
2.5% CSRP
5% CSRP
10% CSRP
15% CSRP
Lactobacillus acidophilus
0.85 ± 0.02 c
1.28 ± 0.01 b
1.73 ± 0.01 a
1.72 ± 0.01 a
Bifidobacterium longum
2.16 ± 0.03 b
2.16 ± 0.01 b
2.34 ± 0.02 a
1.62 ± 0.07 c
Note: values are Probiotic value ± SD of CSRP, because a, b, c, d, and e are in the same row, the
average values of dierent leers were signicantly dierent when p < 0.05, n = 3.
3.3. Evaluation of Prebiotic Activity In Vivo
Eects of CSRP Intake on the Composition of Cecal Gut Microbiotas
A high-throughput analysis of the 16S rDNA V3-V4 region of the mice feces revealed
a total of 1,844,690 sequences and 477 operational taxonomic units (OUTs). The data were
stored in NCBI sequence reading les with the login number SRP463499. To assess the
richness and diversity of the microbial community, the Chao1 index and Ace index were
used to analyze richness, while the Shannon index and Simpson index were used to de-
scribe diversity and evenness. Coverage was used to represent the coverage of the micro-
bial community [20]. The results, shown in Table 4, indicated that as the concentration of
CSRPs increased, the index values initially increased and then decreased. Among the
groups, the M group exhibited the highest richness, while the H group had the highest
diversity.
Table 4. α-diversity index of gut microbiota in each group.
Sample
Richness
Diversity
Coverage
Chao1
Ace
Shannon
Simpson
CK
288.26 ± 26.66
286.53 ± 27.36
3.40 ± 0.39
0.10 ± 0.04
0.999
L
312.61 ± 19.09
306.36 ± 21.03
3.38 ± 0.36
0.08 ± 0.02
0.999
Figure 3.
(
a
) Effects of different concentrations of CSRPs on growth density of Lactobacillus acidophilus;
(
b
) effects of different concentrations of CSRPs on growth density of Bifidobacterium longum.
** p< 0.01
,
*** p< 0.001). (10% CSRP can promote the growth of probiotics).
Table 3. Effects of CSRPs on PI values of probiotics.
2.5% CSRP 5% CSRP 10% CSRP 15% CSRP
Lactobacillus acidophilus 0.85 ±0.02 c 1.28 ±0.01 b 1.73 ±0.01 a 1.72 ±0.01 a
Bifidobacterium longum 2.16 ±0.03 b 2.16 ±0.01 b 2.34 ±0.02 a 1.62 ±0.07 c
Note: values are Probiotic value
±
SD of CSRP, because a, b, and c are in the same row, the average values of
different letters were significantly different when p< 0.05, n= 3.
3.3. Evaluation of Prebiotic Activity In Vivo
Effects of CSRP Intake on the Composition of Cecal Gut Microbiotas
A high-throughput analysis of the 16S rDNA V3-V4 region of the mice feces revealed
a total of 1,844,690 sequences and 477 operational taxonomic units (OUTs). The data were
stored in NCBI sequence reading files with the login number SRP463499. To assess the
richness and diversity of the microbial community, the Chao1 index and Ace index were
used to analyze richness, while the Shannon index and Simpson index were used to describe
diversity and evenness. Coverage was used to represent the coverage of the microbial
community [
20
]. The results, shown in Table 4, indicated that as the concentration of CSRPs
increased, the index values initially increased and then decreased. Among the groups, the
M group exhibited the highest richness, while the H group had the highest diversity.
Table 4. α-diversity index of gut microbiota in each group.
Sample Richness Diversity Coverage
Chao1 Ace Shannon Simpson
CK 288.26 ±26.66 286.53 ±27.36 3.40 ±0.39 0.10 ±0.04 0.999
L 312.61 ±19.09 306.36 ±21.03 3.38 ±0.36 0.08 ±0.02 0.999
M 316.24 ±38.84 308.36 ±39.18 3.38 ±0.81 0.06 ±0.36 0.999
H 284.81 ±17.29 284.57 ±17.29 2.84 ±0.28 0.18 ±0.05 0.998
Data are presented as mean ±SD (n= 6) and compared with control group.
Nutrients 2023,15, 4843 8 of 12
Additionally, a principal component analysis (PCA) (Figure 4A) and the unweighted
UNIFRAC distance were employed to calculate a Principal Coordinate Analysis (PCoA)
(Figure 4B) to evaluate the community composition. The results demonstrated a significant
separation between the groups, indicating a significant difference in gut microbiota with
or without the intake of CSRPs. The analysis of the gut microbiota composition at the
phylum level revealed that the dominant microbial phyla were Bacteroidota, Firmicutes,
Actinobacteriota, Desulfobacterota, and Campilobacterota (Figure 4C). At the genus level,
the dominant flora included Muribaculaceae,Alloprevotella,Dubosiella,Bacteroides,Lactobacil-
lus,Lachnospiraceae,Faecalibaculum,Bifidobacterium,Eubacterium, and others. With increasing
concentrations of CSRPs, the relative abundance of beneficial bacteria such as Dubosiella,
Lactobacillus,Faecalibaculum, and Bifidobacterium increased, while the growth of Bacteroides
and Enterorhabdus was inhibited (Figure 4D).
A cluster analysis revealed high composition similarity at the genus level within each
group, with some genera showing high similarity and others displaying significant differ-
ences. As the concentrations of CSRPs increased, the differences between the groups became
more pronounced (Figure 4E). A significant difference test analysis of gut microbiota at
the genus level was conducted between the groups, and the results indicated significant
differences in the bacterial composition of gut microbiota at the genus level in mice re-
ceiving different intragastric doses. This included Dubosiella,Bacteroides,Faecalibaculum,
Bifidobacterium,Trichospirillum, Eubacterium,Segmented filamentous bacteria,Faecalibacterium,
and Enterorhabdus (one-way ANOVA) (Figure 4F).
Nutrients 2023, 15, x FOR PEER REVIEW 8 of 12
M 316.24 ± 38.84 308.36 ± 39.18 3.38 ± 0.81 0.06 ± 0.36 0.999
H 284.81 ± 17.29 284.57 ± 17.29 2.84 ± 0.28 0.18 ± 0.05 0.998
Data are presented as mean ± SD (n = 6) and compared with control group.
Additionally, a principal component analysis (PCA) (Figure 4A) and the unweighted
UNIFRAC distance were employed to calculate a Principal Coordinate Analysis (PCoA)
(Figure 4B) to evaluate the community composition. The results demonstrated a signifi-
cant separation between the groups, indicating a significant difference in gut microbiota
with or without the intake of CSRPs. The analysis of the gut microbiota composition at
the phylum level revealed that the dominant microbial phyla were Bacteroidota, Firmicu-
tes, Actinobacteriota, Desulfobacterota, and Campilobacterota (Figure 4C). At the genus
level, the dominant flora included Muribaculaceae, Alloprevotella, Dubosiella, Bacteroides,
Lactobacillus, Lachnospiraceae, Faecalibaculum, Bifidobacterium, Eubacterium, and others. With
increasing concentrations of CSRPs, the relative abundance of beneficial bacteria such as
Dubosiella, Lactobacillus, Faecalibaculum, and Bifidobacterium increased, while the growth of
Bacteroides and Enterorhabdus was inhibited (Figure 4D).
Figure 4. Cont.
Nutrients 2023,15, 4843 9 of 12
Nutrients 2023, 15, x FOR PEER REVIEW 9 of 12
Figure 4. Effects of CSRPs on gut microbiota in mice. Control group was intragastrically adminis-
tered with normal saline, L group was intragastrically administered with low-dose CSRP, M group
was intragastrically administered with medium-dose CSRP, and H group was intragastrically ad-
ministered with high-dose CSRP. (A) Principal component analysis (PCA); (B) calculating principal
component analysis based on unweighted UNIFRAC distance (PCoA); (C) bacteria relative abun-
dance at phylum level (%); (D) bacteria relative abundance at genus level (%); (E) bacterial clustering
heat map; (F) significance test of difference between groups, * p < 0.05, ** p < 0.01.
4. Discussion
Physicochemical changes in prebiotics can have a significant impact on the composi-
tion, digestive function, and immune response of the gastrointestinal microbiota. Prebiot-
ics are selectively utilized by beneficial microorganisms in the host and have various
health benefits. They can enhance resistance to pathogens, regulate immune function, im-
prove mineral absorption, promote intestinal function, affect metabolism, and contribute
to satiety [21]. Probiotics, on the other hand, play a role in immune regulation, the pro-
duction of organic acids and antimicrobial compounds, interactions with resident micro-
biota, interactions with the host, improvement in intestinal barrier integrity, and enzyme
formation [2]. Stellariae Radix is a traditional Chinese medicine, but its polysaccharides
have been rarely studied. Our research has demonstrated that Stellariae Radix contains
abundant water-soluble polysaccharides and possesses free radical scavenging effects
[16]. In this study, we found that CSRPs can promote the growth of Lactobacillus acidophilus
and Bifidobacterium longum in an MRS medium. This was evident from the significant in-
crease in the sugar consumption and clump count of the two probiotics, as well as the
reduction in pH in the sugar-free MRS medium. The PI values also indicated that the ad-
dition of 10% crude polysaccharides in the medium had the most pronounced effect on
promoting the growth of probiotics. Therefore, our in vitro evaluation of prebiotic activity
showed that CSRPs exhibited stronger prebiotic activity than inulin in promoting the
growth of these two probiotics. This provides a theoretical basis for the potential devel-
opment of Stellariae Radix as a prebiotic drug in the fields of food, healthcare products,
and animal feed.
The gut microbiota plays a crucial role in human health and is associated with vari-
ous diseases. The treatment and prevention of many intestinal-related diseases using pro-
biotics and prebiotics involve modulating the microbiota and/or its functions, such as in-
creasing the proportion of probiotics in the intestine and regulating the intestinal micro-
environment [22]. In the intestine, Firmicutes and Bacteroidetes are the dominant bacterial
phyla, and their proportion changes have been studied in relation to diseases such as obe-
sity. Obesity is characterized by an increase in the proportion of Firmicutes and Bac-
teroidetes in the gut microbiota [23]. In our study, the results of the alpha diversity anal-
ysis showed that the low-dose and medium-dose groups increased the diversity and rich-
Figure 4.
Effects of CSRPs on gut microbiota in mice. Control group was intragastrically administered
with normal saline, L group was intragastrically administered with low-dose CSRP, M group was in-
tragastrically administered with medium-dose CSRP, and H group was intragastrically administered
with high-dose CSRP. (
A
) Principal component analysis (PCA); (
B
) calculating principal component
analysis based on unweighted UNIFRAC distance (PCoA); (
C
) bacteria relative abundance at phy-
lum level (%); (
D
) bacteria relative abundance at genus level (%); (
E
) bacterial clustering heat map;
(F) significance test of difference between groups, * p< 0.05, ** p< 0.01.
4. Discussion
Physicochemical changes in prebiotics can have a significant impact on the composi-
tion, digestive function, and immune response of the gastrointestinal microbiota. Prebiotics
are selectively utilized by beneficial microorganisms in the host and have various health
benefits. They can enhance resistance to pathogens, regulate immune function, improve
mineral absorption, promote intestinal function, affect metabolism, and contribute to
satiety [21].
Probiotics, on the other hand, play a role in immune regulation, the produc-
tion of organic acids and antimicrobial compounds, interactions with resident microbiota,
interactions with the host, improvement in intestinal barrier integrity, and enzyme for-
mation [
2
]. Stellariae Radix is a traditional Chinese medicine, but its polysaccharides
have been rarely studied. Our research has demonstrated that Stellariae Radix contains
abundant water-soluble polysaccharides and possesses free radical scavenging effects [
16
].
In this study, we found that CSRPs can promote the growth of Lactobacillus acidophilus and
Bifidobacterium longum in an MRS medium. This was evident from the significant increase
in the sugar consumption and clump count of the two probiotics, as well as the reduction
in pH in the sugar-free MRS medium. The PI values also indicated that the addition of 10%
crude polysaccharides in the medium had the most pronounced effect on promoting the
growth of probiotics. Therefore, our
in vitro
evaluation of prebiotic activity showed that
CSRPs exhibited stronger prebiotic activity than inulin in promoting the growth of these
two probiotics
. This provides a theoretical basis for the potential development of Stellariae
Radix as a prebiotic drug in the fields of food, healthcare products, and animal feed.
The gut microbiota plays a crucial role in human health and is associated with var-
ious diseases. The treatment and prevention of many intestinal-related diseases using
probiotics and prebiotics involve modulating the microbiota and/or its functions, such
as increasing the proportion of probiotics in the intestine and regulating the intestinal
microenvironment [
22
]. In the intestine, Firmicutes and Bacteroidetes are the dominant
bacterial phyla, and their proportion changes have been studied in relation to diseases
such as obesity. Obesity is characterized by an increase in the proportion of Firmicutes
and Bacteroidetes in the gut microbiota [
23
]. In our study, the results of the alpha diversity
analysis showed that the low-dose and medium-dose groups increased the diversity and
richness of the gut microbiota compared to the control group, while the high-dose group
reduced them. At the phylum level, the low-dose and medium-dose groups reduced the
Nutrients 2023,15, 4843 10 of 12
abundance of Firmicutes and increased the abundance of Bacteroidetes, thereby reducing
the Firmicutes/Bacteroidetes ratio. This suggests that low and medium doses of CSRPs in
mice may have a preventive and therapeutic effect on obesity by modulating the intestinal
environment. Firmicutes and Bacteroidetes represent both beneficial and potentially harm-
ful bacteria, and their relative abundance lies between the optimal levels [
24
]. Therefore,
the reduction in the diversity and richness of the gut microbiota observed in the high-
dose group may be due to a decrease in the abundance or even the elimination of certain
harmful bacteria.
In the analysis of bacterial composition at the genus level, the hypothesis was con-
firmed. With an increase in the concentration of crude polysaccharides, the relative abun-
dance of Dubosiella,Faecalibaculum, Bifidobacterium, and Lactobacillus significantly increased,
while harmful bacteria such as Enterorhabdus decreased significantly. It has been reported
that Dubosiella,Bifidobacterium, and Lactobacillus are positively correlated with preventing
and improving obesity and related diseases [
25
]. Zhu et al. found that Dubosiella can
improve abnormal indexes in obese mice induced by a high-fat diet, reduce LDL-C, TG,
and body weight, improve the digestion and absorption ability of glucose in mice, and
reduce the expression levels of lipid metabolism genes such as CD36, FASN, and PPAR
γ
in the liver, thereby alleviating the disorder of lipid metabolism caused by obesity [
24
].
Additionally, Dubosiella can increase the abundance of beneficial microorganisms (Bifidobac-
terium and Lactobacillus) in obese mice, improve the disorder of intestinal flora structure
caused by a high-fat diet, improve intestinal metabolism and immunity, and reduce the
occurrence of bacterial infections and other diseases. Numerous studies have also found
that the content of Dubosiella increases during the treatment of obesity [
26
–
32
]. It has been
proven to improve the metabolism of the body, normalize blood lipid metabolism, and
increase the content of short-chain fatty acids (SCFAs) such as butyrate. Butyrate has
the ability to promote the conversion of white adipose tissue (WAT) to brown adipose
tissue (BAT). WAT stores energy, while BAT uses energy for heat production, resulting in
increased energy consumption by the host and the degradation of fat by regulating the
host’s TCA cycle. In this study, with the increase in CSRP concentration, the abundance of
Lactobacillus,Bifidobacterium, and Dubosiella in the intestinal tract of the mice significantly
increased. Therefore, this further supports the speculation that CSRPs can regulate the gut
microbiota and potentially treat obesity.
Furthermore, certain intestinal microorganisms (Bacteroides,Prevotella, and Lactobacil-
lus) play a crucial role in hydrolyzing polysaccharides into soluble cytokines [
33
]. The
microbial fermentation of dietary carbohydrates, including prebiotics, promotes the pro-
duction of SCFAs [
34
]. SCFAs are well-known signaling molecules. Firstly, SCFAs create
an acidic environment in the colon, inhibiting the growth of pathogens. Secondly, SC-
FAs can regulate the growth and differentiation of intestinal cells and provide them with
energy [35,36]
. Studies have shown that Bacteroides can stimulate the production of propi-
onic acid, which limits acetate synthesis and reduces SCFA synthesis, resulting in a negative
correlation between SCFA content and the number of Bacteroides in the intestine [
37
]. In
this study, compared to the control group, the treatment groups significantly reduced the
relative abundance of Bacteroides in the intestine (p< 0.05) and inhibited its growth with
increasing dosage. This indicates that crude polysaccharides can effectively inhibit the
growth of Bacteroides, providing a theoretical basis for further studying the effects of SRPs
on the SCFA content in the intestine.
5. Conclusions
In summary, this study provides a solid foundation for understanding the prebiotic
effect of CSRPs. It demonstrates that CSRPs have the ability to enhance the growth of
beneficial bacteria in mice, surpassing the prebiotic activity of inulin
in vitro
. Additionally,
CSRPs have the potential to regulate the abundance and diversity of gut microbiota in mice.
These findings suggest that a CSRP could be utilized as a prebiotic to modulate the gut
microbiota, paving the way for the further exploration and utilization of Stellariae Radix
Nutrients 2023,15, 4843 11 of 12
as a valuable medicinal resource. This study provides a crucial theoretical basis for future
research on CSRPs.
6. Patents
Peng, L., Wang, H., Song, L., Feng, L., Niu, P.L., Li, Z.K., Li, Y.Q., Li, H.S. & Wu, W. et al.
A crude polysaccharide of Stellariae Radix with prebiotic activity, and its preparation
method [P]. China: CN115677874A, 3 February 2023.
Author Contributions:
H.W.: performed the study and analyzed the results. H.L.: data interpretation
and manuscript revision. Z.L.: conceptualization and data interpretation. L.F.: performed the study
and analyzed the results. L.P.: designed the study, contributed to the concept generation, and
wrote and revised the manuscript. All authors have read and agreed to the published version of
the manuscript.
Funding:
This study was supported by the Key Research and Development Program of Ningxia
(No. 2021BEG02042).
Institutional Review Board Statement:
The investigation complies with the Guide for Care and Use
of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23,
revised 2011) and was approved by the Experimental Animal Ethics Committee of Ningxia Medical
University (Production license number: SCXK (Ning) 2020-0001. Usage license number: SYXK(Ning)
2020-0001).
Data Availability Statement:
The original contributions presented in the study are included in the
article, and further inquiries can be directed to the corresponding authors.
Conflicts of Interest: The authors declare no conflict of interest.
References
1.
Altermann, E.; Russell, W.M.; Azcarate-Peril, M.A.; Barrangou, R.; Buck, B.L.; McAuliffe, O.; Souther, N.; Dobson, N.; Duong, T.;
Callanan, M. Complete genome sequence of the probiotic lactic acidbacterium Lactobacillus acidophilus NCFM. Proc. Natl. Acad.
Sci. USA 2005,102, 3906–3912. [CrossRef] [PubMed]
2.
Xu, Y.; Wu, Y.-J.; Sun, P.-L.; Zhang, F.-M.; Linhardt, R.-J.; Zhang, A.-Q. Chemically modified polysaccharides: Synthesis,
characterization, structure activity relationships of action. Int. J. Biol. Macromol. 2019,132, 970–977. [CrossRef] [PubMed]
3.
Wang, Z.; Lu, C.; Wu, C.; Xu, M.; Kou, X.; Kong, D.; Jing, G. Polysaccharide of Boschniakia rossica induces apoptosis on laryngeal
carcinoma Hep2 cells. Gene 2014,536, 203–206. [CrossRef]
4.
Yang, Y.; Ji, J.; Di, L.; Li, J.; Hu, L.; Qiao, H.; Wang, L.; Feng, Y. Resource, chemical structure and activity of natural polysaccharides
against alcoholic liver damages. Carbohydr. Polym. 2020,241, 116355. [CrossRef] [PubMed]
5.
Xu, S.-Y.; Aweya, J.J.; Li, N.; We, H.; Sun, C.; Li, J.; Jiang, Y. Microbial catabolism of Porphyra haitanensis polysaccharides by human
gut microbiota. Food Chem. 2019,289, 177–186. [CrossRef] [PubMed]
6.
Chen, X.; Fang, Y.; Nishinari, K.; We, H.; Sun, C.; Li, J.; Jiang, Y. Physicochemical characteristics of polysaccharide conjugates
prepared from fresh tea leaves and their improving impaired glucose tolerance. Carbohydr. Polym. 2014,112, 77–84. [CrossRef]
7.
Chen, R.; Liu, B.; Wang, X.; Chen, K.; Zhang, K.; Zhang, L.; Fei, C.; Wang, C.; Liu, Y.; Xue, F. Effects of polysaccharide from
Pueraria lobata on gut microbiota in mice. Int. J. Biol. Macromol. 2020,158, 740–749. [CrossRef]
8.
Porter, N.T.; Martens, E.C. The Critical Roles of Polysaccharides in Gut Microbial Ecology and Physiology. Annu. Rev. Microbiol.
2017,71, 349–369. [CrossRef]
9.
Zhu, W.; Zhou, S.; Liu, J.; McLean, R.; Chu, W. Prebiotic, immuno-stimulating and gut microbiota-modulating effects of Lycium
barbarum polysaccharide. Biomed. Pharmacother. 2020,121, 109591. [CrossRef]
10.
Xie, S.-Z.; Liu, B.; Ye, H.-Y.; Li, Q.-M.; Pan, L.-H.; Zha, X.-Q.; Liu, J.; Duan, J.; Luo, J.-P. Dendrobium huoshanense polysaccharide
regionally regulates intestinal mucosal barrier function and intestinal microbiota in mice. Carbohydr. Polym.
2019
,206, 149–162.
[CrossRef]
11. Fang, W.P.; Zhang, Z.R. Flora Reipublicae Popularis Sinicae; Science Press: Beijing, China, 2004.
12. China Pharmacopoeia; National Pharmacopoeia Committee: Beijing, China, 2020; pp. 330–331.
13.
Li, Z.K.; Wang, H.; Song, L.; Feng, L.; Li, Y.Q.; Yang, Y.; Peng, L. Origin Characteristics and Correlation Analysis of Stellarae Radix
Based on Inorganic Elements and Effective Components. Chin. J. Mod. Appl. Pharm. 2023,40, 894–902.
14.
Song, L.; Wang, H.; Li, Z.K.; Liu, D.H.; Peng, L. Optimization of Supercritical CO
2
Extraction Process and Component Analysis of
Extract from Stellariae Radix. Chin. J. Mod. Appl. Pharm. 2022,39, 2498–2504.
15.
Li, Z.K.; Ma, L.; Bai, M.S.; Huang, Y.X.; Song, L.; Wang, H.; Feng, L.; Peng, L. Infrared fingerprint of Ningxia Stellariae Radix
based on chemometrics and discrimination analysis of counterfeit products. Mod. Tradit. Chin. Med. Mater. Medica-World Sci.
Technol. 2023,25, 175–183.
Nutrients 2023,15, 4843 12 of 12
16.
Wang, H.; Peng, L.; Song, L.; Feng, L.; Li, Z.K.; Li, Y.Q.; Gao, T. Optimization of Ultrasonic-assisted Extraction Process and
Analysis of Antioxidant Activity of Polysaccharide from Stellariae Radix. Food Ind. Sci. Technol. 2023, 1–12. [CrossRef]
17.
Zhang, T.Y.; Zhang, X.Y.; Wang, D.P.; Xu, X.X. The Effect of Polysaccharide of Poriacocos on Key Metabolites of Bifidobacterium
BB-12. Sci. Technol. Food Ind. 2022,43, 24–33.
18.
Zwietering, M.; Jongenburger, I.; Rombouts, F.; Riet, K. Modeling of the bacterial growth curve. Appl. Environ. Microbiol.
1990
,56,
1875–1881. [CrossRef]
19.
Su, J.; Su, L.; Li, D.; Shuai, O.; Zhang, Y.; Liang, H.; Jiao, C.; Xu, Z.; Lai, Y.; Xie, Y. Antitumor Activity of Extract from the
Sporoderm-Breaking Spore of Ganoderma lucidum: Restoration on Exhausted Cytotoxic T Cell with Gut Microbiota Remodeling.
Front. Immunol. 2018,9, 1765. [CrossRef]
20.
Peng, L.; Wang, H.; Song, L.; Feng, L.; Niu, P.L.; Li, Z.K.; Li, Y.Q.; Li, H.S.; Wu, W. A Crude Polysaccharide of Stellariae Radix with
Prebiotic Activity, and Its Preparation. Method. Patent CN115677874A, 3 February 2023.
21.
Liu, B.; Liu, T.T.; Zhang, X.K.; Wang, T.; Zhang, C.X.; Li, M.; Zheng, X.Y.; Wang, D.P.; Zhang, D.Z.; Meng, X.J. Research progress of
prebiotics and their applications. Food Med. 2021,23, 485–492.
22.
Huang, L.; Ke, H.; Chen, G.H.; Zheng, Y. Preparation and quality analysis of Polygonatum polysaccharide prebiotic cake. Food
Ind. 2020,41, 90–94.
23.
Zhang, S.; Dang, Y. Roles of gut microbiota and metabolites in overweight and obesity of children. Front. Endocrinol.
2022
,
13, 994930. [CrossRef]
24.
Seo, K.H.; Kim, D.H.; Jeong, D. Chardonnay grape seed flour supplemented diets alter intestinal microbiota in diet-induced obese
mice. J. Food Biochem. 2017,41, e12396. [CrossRef]
25.
Zhu, S.L.; Chen, Y.Q.; Jiang, X. Application of Dunaliella in Preventing and Improving Obesity and Related. Diseases. Patent
CN111686133A, 22 September 2020.
26.
Zhou, X.; Ma, L.; Dong, L.; Li, D.; Chen, F.; Hu, X. Bamboo shoot dietary fiber alleviates gut microbiota dysbiosis and modulates
liver fatty acid metabolism in mice with high-fat diet-induced obesity. Front. Nutr. 2023,10, 1161698. [CrossRef] [PubMed]
27.
Rodríguez-Daza, M.C.; Roquim, M.; Dudonné, S.; Pilon, G.; Levy, E.; Marette, A.; Roy, D.; Desjardins, Y. Berry Polyphenols and
Fibers Modulate Distinct Microbial Metabolic Functions and Gut Microbiota Enterotype-Like Clustering in Obese Mice. Front.
Microbiol. 2020,11, 2032. [CrossRef]
28.
Gao, J.; Ma, L.; Yin, J.; Liu, G.; Ma, J.; Xia, S.; Gong, S.; Han, Q.; Li, T.; Chen, Y. Camellia (Camellia oleifera bel.) seed oil reprograms
gut microbiota and alleviates lipid accumulation in high fat-fed mice through the mTOR pathway. Food Funct.
2022
,13, 4977–4992.
[CrossRef]
29.
Yang, K.; Jian, S.; Wen, C.; Guo, D.; Liao, P.; Wen, J.; Kuang, T.; Han, S.; Liu, Q.; Deng, B. Gallnut Tannic Acid Exerts Anti-stress
Effects on Stress-Induced Inflammatory Response, Dysbiotic Gut Microbiota, and Alterations of Serum Metabolic Profile in Beagle
Dogs. Front. Nutr. 2022,9, 847966. [CrossRef]
30.
Qiu, X.; Macchietto, M.G.; Liu, X.; Lu, Y.; Ma, Y.; Guo, H.; Saqui-Salces, M.; Bernlohr, D.A.; Chen, C.; Shen, S.; et al. Identification
of gut microbiota and microbial metabolites regulated by an antimicrobial peptide lipocalin 2 in high fat diet-induced obesity. Int.
J. Obes. 2021,45, 143–154. [CrossRef]
31.
Zhu, Z.; Huang, R.; Huang, A.; Wang, J.; Liu, W.; Wu, S.; Chen, M.; Chen, M.; Xie, Y.; Jiao, C.; et al. Polysaccharide from Agrocybe
cylindracea prevents diet-induced obesity through inhibiting inflammation mediated by gut microbiota and associated metabolites.
Int. J. Biol. Macromol. 2022,209 Pt A, 1430–1438. [CrossRef]
32.
Deng, M.; Zhang, S.; Dong, L.; Huang, F.; Jia, X.; Su, D.; Chi, J.; Muhammad, Z.; Ma, Q.; Zhao, D.; et al. Shatianyu (Citrus
grandis L. Osbecontrol) Flavonoids and Dietary Fiber in Combination Are More Effective Than Individually in Alleviating High-
Fat-Diet-Induced Hyperlipidemia in Mice by Altering Gut Microbiota. J. Agric. Food Chem.
2022
,70, 14654–14664. [CrossRef]
[PubMed]
33.
Poeker, S.A.; Geirnaert, A.; Berchtold, L.; Greppi, A.; Krych, L.; Steinert, R.E.; de Wouters, T.; Lacroix, C. Understanding the
prebiotic potential of different dietary fibers using an in vitro continuous adult fermentation model (PolyFermS). Sci. Rep. 2018,
8, 4318. [CrossRef]
34.
Yadav, M.K.; Kumari, I.; Singh, B.; Sharma, K.K.; Tiwari, S.K. Probiotics, prebiotics and synbiotics: Safe options for next-generation
therapeutics. Appl. Microbiol. Biotechnol. 2022,106, 505–521. [CrossRef]
35.
Cummings, J.H.; Pomare, E.W.; Branch, W.J.; Naylor, C.P.; Macfarlane, G.T. Short chain fatty acids in human large intestine, portal,
hepatic and venous blood. Gut 1987,28, 1221. [CrossRef] [PubMed]
36.
Huang, Q.; Cai, G.; Liu, T.; Liu, Z. Relationships Among Gut Microbiota, Ischemic Stroke and Its Risk Factors: Based on Research
Evidence. Int. J. Gen. Med. 2022,15, 2003–2023. [CrossRef] [PubMed]
37.
Gu, W.; Wang, Y.; Zeng, L.; Dong, J.; Bi, Q.; Yang, X.; Che, Y.; He, S.; Yu, J. Polysaccharides from Polygonatum kingianum improve
Glccose and lipid metabolism in rats fed a high fat diet. Biomed. Pharmacother. 2020,125, 109910. [CrossRef] [PubMed]
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