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Observational Study
1
Medicine®
Network pharmacology and molecular docking
reveal potential mechanisms of ginseng in the
treatment of diabetes mellitus-induced erectile
dysfunction and asthenospermia
Liming Liu, MMa,b, Yuanfeng Zhang, MDc,d, Jiashu Yang, MMb, Wenfang Chen, BDe, Kaijian Lan, MDd,
Yibo Shi, MDc, Xiaogang Zhang, MMf, Xiping Xing, MMg,*
Abstract
Diabetes mellitus (DM) is a chronic metabolic disease that predisposes to chronic damage and dysfunction of various organs,
including leading to erectile dysfunction (ED) and asthenospermia. Literature suggests that ginseng plays an important role in
the treatment and management of DM. Ginseng may have a therapeutic effect on the complications of DM-induced ED and
asthenospermia. The study aimed to explore the mechanisms of ginseng in the treatment of DM-induced ED and asthenospermia
following the Traditional Chinese Medicine (TCM) theory of “treating different diseases with the same treatment.” This study used
network pharmacology and molecular docking to examine the potential targets and pharmacological mechanism of Ginseng for
the treatment of DM-induced ED and asthenospermia. The chemical ingredients and targets of ginseng were acquired using the
Traditional Chinese Medicine Systems Pharmacology database and analysis platform. The targets of DM, ED, and asthenospermia
were extracted with the GeneCards and Online Mendelian Inheritance in Man databases. A protein–protein interaction network
analysis was constructed. The Metascape platform was applied for analyzing the gene ontology and Kyoto Encyclopedia of
Genes and Genomes pathways. AutoDock Vina was used to perform molecular docking. Network pharmacology revealed that
the main active components of the target of action were kaempferol, beta-sitosterol, ginsenoside rh2, stigmasterol, and fumarine.
Core targets of the protein–protein interaction network included TNF, IL-1β, AKT1, PTGS2, BCL2, and JUN. Kyoto Encyclopedia
of Genes and Genomes enrichment analysis showed that they were mainly involved in AGE-RAGE signaling pathway in diabetic
complications, TNF signaling pathway, Lipid and atherosclerosis. The interactions of core active components and targets were
analyzed by molecular docking. Ginseng may play a comprehensive therapeutic role in the treatment of DM-induced ED and
asthenospermia through “multicomponent, multi-target, and multi-pathway” biological mechanisms such as inflammation and
oxidative stress.
Abbreviations : AGEs = advanced glycation end products, ART = assisted reproductive technologies, BCbetweenness
centralityBCLb-cell lymphomaBP = biological processes, CC = cell components, COX-2 = cyclooxygenase-2, DC = degree
centrality, DL = drug-like properties, DM = diabetes mellitus, DMED = diabetes mellitus-induced erectile dysfunction, ED =
erectile dysfunction, GO = gene ontology, KEGG = Kyoto Encyclopedia of Genes and Genomes, LPS = lipopolysaccharide, MF =
molecular functions, OB = oral bioavailability, OMIMOnline Mendelian Inheritance in ManPPI = protein–protein interaction, TCM =
traditional Chinese medicine, TCMSP = Traditional Chinese Medicine Systems Pharmacology, TLR4 = Toll-like receptor 4.
Keywords: asthenospermia, diabetes mellitus, erectile dysfunction, network pharmacology, treating different diseases with the
same treatment
XZ and XX contributed to this article equally.
This study was supported by Gansu Provincial Key Laboratory of Chinese
Medicine for Prevention and Treatment of Chronic Diseases Open Fund (Grant
No. GSMBKY2015-13); Natural Science Foundation of Gansu Province (Grant
No. 23JRRA1198); Guidance Plan for Science and Technology Development
Projects in Lanzhou City (Grant No. 2023-ZD-102); Intra-Hospital Project of the
Affiliated Hospital of Gansu University of Chinese Medicine (Grant No. gzfy-
2019-07); The Medical Science and Technology Program of Shantou (Grant No.
2021-3-44).
The authors have no conflicts of interest to disclose.
The datasets generated during and/or analyzed during the current study are
available from the corresponding author on reasonable request.
a Department of Andrology, Xi’an Hospital of Traditional Chinese Medicine,Xi’an,
P. R. China, b School of Integrated Chinese and Western Medicine, Gansu
University of Chinese Medicine, Lanzhou, P. R. China, c Department of Urology,
Key Laboratory of Urological Disease of Gansu Province, Clinical Center of Gansu
Province for Nephron-Urology, Lanzhou University Second Hospital, Lanzhou,
P. R. China, d Department of Urology, Shantou Central Hospital, Shantou, P. R.
China, e The Second Clinical Medical College, Lanzhou University, Lanzhou, P. R.
China, f Department of Cardiology, Affiliated Hospital of Gansu Medical College,
Pingliang, P. R. China, g Department of Urology and Andrology, Affiliated Hospital
of Gansu University of Chinese Medicine, Lanzhou, P. R. China.
* Correspondence: Xiping Xing, Affiliated Hospital of Gansu University of Chinese
Medicine, Chengguan District, Lanzhou, Gansu Province, 730020, China (email:
xxp214@126.com).
Copyright © 2024 the Author(s). Published by Wolters Kluwer Health, Inc.
This is an open-access article distributed under the terms of the Creative
Commons Attribution-Non Commercial License 4.0 (CCBY-NC), where it is
permissible to download, share, remix, transform, and buildup the work provided
it is properly cited. The work cannot be used commercially without permission
from the journal.
How to cite this article: Liu L, Zhang Y, Yang J, Chen W, Lan K, Shi Y, Zhang
X, Xing X. Network pharmacology and molecular docking reveal potential
mechanisms of ginseng in the treatment of diabetes mellitus-induced erectile
dysfunction and asthenospermia. Medicine 2024;103:34(e39384).
Received: 18 January 2024 / Received in final form: 30 July 2024 / Accepted: 31
July 2024
http://dx.doi.org/10.1097/MD.0000000000039384
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Liu et al. • Medicine (2024) 103:34 Medicine
1. Introduction
Diabetes mellitus (DM) is a metabolic disease characterized
by chronic hyperglycemia, insufcient insulin secretion and/or
dysfunction, which results in severe disturbances in carbohy-
drate, fat, and protein metabolism, as well as chronic damage,
dysfunction and failure of various organs.[1] According to the
International Diabetes Federation, the number of people with
DM will increase to 693 million by 2045, which represents
about 10% of the global population.[2] In addition to induc-
ing complications such as cardiovascular disease, nephropathy
and retinopathy, DM can lead to erectile dysfunction (ED),
and asthenospermia, which can be a sign of impaired health
in men.[3–5] Diabetes-induced ED is a common complication in
men with DM, which is mainly characterized by the inability
to achieve or maintain a sustained and adequate erection for
satisfactory sexual intercourse. DM-induced ED may be caused
by a variety of factors such as oxidative stress, inammation,
advanced glycation end products (AGEs), endothelial dysfunc-
tion, and vascular nerve damage.[6] Clinical treatments mainly
include phosphodiesterase type 5 inhibitors, penile cavernous
injections and surgery, however, they all have certain limitations
and adverse effects, and their use is restricted.[7] In addition,
male DM patients are also susceptible to the complication of
asthenospermia, which is mainly characterized by the propor-
tion of forward-moving spermatozoa in the semen being <32%
or the ratio of forward-moving to non-forward-moving sper-
matozoa being <40%.[8] It was found that the pathogenesis of
DM-induced asthenospermia may be closely related to oxidative
stress and inammation caused by DM.[9] Empirical pharma-
cological treatments, assisted reproductive technologies (ART)
and surgical treatments are mainly used clinically to improve
fertility rates at this stage.[10,11] However, the efcacy of existing
drugs is not signicant, and the success rate of ART is low and
expensive, and therefore their clinical promotion is relatively
limited.[12]
Traditional Chinese medicine (TCM) has many advantages in
preventing the complications of DM. It has been receiving more
and more attention because it has less toxicity and side effects,
improves symptoms signicantly, and can make up for the inad-
equacy of Western medicines.[13,14] Ginseng (Panax ginseng C.A.
Meyer) is a valuable Chinese herb with a wide range of appli-
cations and a long history. Ginseng is listed as a top quality in
the Shennong Herbal Classic, which is 1 of the 4 classic works
of TCM and the earliest work on Chinese medicine.[15] Modern
pharmacological studies have proved that ginseng is rich in a
variety of proteins, polysaccharides and saponins, which have
antioxidant and anti-inammatory effects.[16] Clinical stud-
ies have found that ginseng shows a better protection against
DM-induced ED. Ginsenosides were demonstrated to improve
erectile function in diabetes mellitus-induced erectile dysfunc-
tion (DMED) rats by alleviating oxidative stress injury.[17] On
the other hand, ginseng is also effective in the treatment of
asthenospermia. It showed that low concentration of ginseno-
side signicantly increased sperm survival rate and fertilization
rate of infertile patients.[18]
“Same treatment for different diseases” indicates the princi-
ple of the application of the same treatment strategy to patients
with different diseases but with the same syndrome.[19] It had
its origin in the spirit of “different treatments for the same dis-
ease” in the “Internal Classics,”[20] and has proven to be effec-
tive in treating a wide range of diseases in subsequent clinical
practice.[21–23] Network pharmacology is a research method that
starts from the holistic and systematic interactions between drug
targets and diseases, which coincides with the “multicomponent–
multi-target” research idea of TCM, and can reveal the complex
mechanism of TCM more comprehensively.[24] Currently, most
of network pharmacology research focuses on single diseases,
with fewer studies on multiple diseases. Due to the systemic and
holistic perspective of network pharmacology, it may serve as
a potential method to explore the underlying mechanisms of
“the same treatment for different diseases.” Molecular dock-
ing is a mature research method based on computer-simulated
interactions between molecules, aimed at predicting the binding
conformation of small molecule ligands with appropriate target
gene binding sites. The application of network pharmacology
and molecular docking techniques can help to provide a better
understanding of the mechanism of action of drugs.
An abundance of literature suggests that ginseng plays an
important role in the treatment and management of DM.[25]
DM-induced ED and asthenospermia may have the same patho-
genesis. Ginseng may increase sperm survival and improve ED,
but the effects of ginseng on DM-induced ED and asthenosper-
mia and their potential mechanisms have not been fully eluci-
dated.[26] In this study, we investigated the mechanism of action
of ginseng in the treatment of DM-induced ED and asthenosper-
mia from the perspective of “same treatment for different dis-
eases” through network pharmacology and molecular docking,
to provide a scientic basis for the clinical application of ginseng.
2. Material and methods
2.1. Screening of drug active components and targets
Oral bioavailability (OB) is one of the most important phar-
macokinetic parameters in absorption, distribution, metabo-
lism, and excretion of drugs. It represents the extent and rate
at which the active ingredients or functional groups of orally
administered drugs are absorbed into the systemic circulation.
OB value serves as a crucial indicator for evaluating whether
a drug can exert its therapeutic effects. A lower OB value sug-
gests a potentially poorer therapeutic effect of the drug. Drug-
like properties (DL) refer to the similarity of a compound to
known drugs. Generally, a lower DL value indicates a lower
likelihood of the compound being an active pharmaceutical
ingredient. Considering that ginseng is administered orally, this
study established thresholds for screening active ingredients of
ginseng based on OB ≥ 30% and DL ≥ 0.18 as suggested by the
Traditional Chinese Medicine Systems Pharmacology (TCMSP)
database (https://old.tcmsp-e.com/tcmsp.php). The active ingre-
dients were predicted using the TCMSP database, and their
target proteins were normalized and converted to gene names
using the UniProt database (https://www.uniprot.org).
2.2. Disease target collection
“Diabetes mellitus,” “Erectile dysfunction,” and “asthenosper-
mia” were utilized as keywords, respectively. The disease targets
of DM, ED, and asthenospermia were collected from GeneCards
(https://www.genecards.org) and the Online Mendelian
Inheritance in Man (http://www.omim.org) databases, and ana-
lyzed using Venny 2.1 software (https://bioinfogp.cnb.csic.es/
tools/venny) for summarization.
2.3. Screening of drug-disease intersection targets
The ginseng targets obtained in “1.1” and “1.2” were matched
with the intersecting targets of DM, ED, and asthenospermia
using the online program Venny 2.1 software. The obtained
targets were considered as potential targets of ginseng for the
treatment of the disease and plotted as Venn diagrams.
2.4. Drug-active components-intersecting target network
construction and analysis
Ginseng active components, targets and disease intersection
targets of DM, ED, and asthenospermia were established as
Network les, and the above elements were also named as
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Liu et al. • Medicine (2024) 103:34 www.md-journal.com
Type les, and the drug-active components-intersection target
network was constructed by using Cytoscape 3.9.1 software,
which was utilized to CytoNCA tool was used for topology
analysis and core components were screened according to the
node degree centrality (DC).
2.5. Construction and analysis of protein–protein
interaction (PPI) network
Upload the drug-disease intersection targets into STRING11.0
(https://string-db.org/), set the species as “Homo sapiens,”
the minimum interaction threshold as “medium condence”
(≥0.400), and the rest of the parameters were default values
to construct the protein–protein interaction (PPI) network. The
PPI network was imported into Cytoscape 3.9.1 software in
TSV format, and the CytoNCA tool of the software was used
to calculate the DC, Betweenness Centrality, and Closeness
Centrality, and the DC value was used to screen out the core
targets.
2.6. GO function and KEGG pathway enrichment analysis
The intersection targets of ginseng targets and disease targets of
DM, ED, and asthenospermia were imported into the Metascape
database (https://metascape.org), and the species was selected as
“sapiens,” and customized analysis was performed to obtain the
data of gene ontology (GO) functional biological process (BP),
molecular function (MF), cellular components (CC) and Kyoto
Encyclopedia of Genes and Genomes (KEGG) pathway data,
which were then analyzed by the microbiology platform (http://
www.bioinformatics.com.cn), and network diagrams were drawn.
2.7. Construction of drug-components-disease-target-
pathway network
To elaborate the mechanism of action of ginseng in the treatment
of DM-induced ED and asthenospermia more clearly and specif-
ically, a Network le was created for each active component in
ginseng and its corresponding target, pathway and corresponding
gene and disease, and the above elements were named as Type
les, which could be imported to obtain the drug-disease-target-
pathway network diagram by using Cytoscape 3.9.1 software.
2.8. Molecular docking
Molecular docking was carried out between the top 5 active
components in the “Drug-Active Components-Intersecting
Targets” network and the core targets obtained from the topol-
ogy analysis of the PPI network. The pdb le of the 3D structure
of the core protein was downloaded from the PDB database; the
mol2 le of the 2D structure of the potential active components
was obtained from the PubChem database. AutoDockTools
and OpenBabel software were used to complete the compound
3D structure transformation, compound and target protein for-
mat conversion, ligand extraction and format conversion, and
proteins were subjected to pre-processing operations such as
dehydrogenation and hydrogenation, and semi-exible docking
was carried out via PyMOL to obtain the evidence of the main
active components of ginseng in the treatment of DM-induced
ED and asthenospermia support.
The schematic illustration of this study is shown in Figure 1.
3. Results
3.1. Screening of ginseng active components and targets
A total of 22 active components of ginseng were obtained after
searching and screening by TCMSP, and the active components
without corresponding targets were deleted. Finally, 17 active
components were obtained and 255 targets corresponding to
ginseng active components were obtained, as shown in Table 1.
After standardized annotation in the UniProt database and dele-
tion of duplicate targets, 88 targets were nally obtained.
3.2. Disease target collection
Nineteen thousand six hundred ninety-one targets associated
with DM, 1804 targets associated with ED, and 5094 targets
associated with asthenospermia were obtained through the
Online Mendelian Inheritance in Man database, respectively.
The above disease targets were summarized using Venny 2.1
software to obtain 20,033 DM targets, 2350 ED targets, and
5476 asthenospermia targets, respectively (Fig. 2A-C), and the
summarized disease targets were intersected by taking the inter-
section of the summarized disease targets using Venny 2.1 soft-
ware to obtain a total of 993 disease targets, which were taken
as the nal source of the candidate disease targets as shown in
Figure 2D.
3.3. Drug-disease intersection targets
The 88 ginseng targets obtained in “1.1” and “1.2” were
imported into the Venny 2.1 software along with 993 intersect-
ing targets for DM, ED, and asthenospermia to obtain drug-
disease intersecting targets and plot the Venny diagram. A total
of 44 drug-disease intersecting targets were obtained (Fig. 2E).
3.4. Drug-active components-intersection target network
The constructed Network le and attribute Type le were
uploaded to the Cytoscape 3.9.1 platform, and the Drug-Active
Components-Intersecting Targets Network graph was drawn
according to its attributes such as nodes and edges (Fig. 3).
The network contains 62 nodes and 114 edges, and the larger
the node degree value, the more important it is in the network.
According to the results of the network topology analysis, the
top 5 active components with degree values were kaempferol,
beta-sitosterol, ginsenoside rh2, stigmasterol, and fumarine
(Table 2), which may be the core components of ginseng for the
treatment of DM-induced ED and asthenospermia.
3.5. PPI networks
The 44 intersecting targets were put through the STRING data-
base to construct a PPI network (Fig. 4A). The network has 44
nodes and 784 edges. The targets in the PPI network were topo-
logically analyzed, and the top 6 core targets were selected based
on the size of DC value (Table 3 and Fig. 4B), which TNF, IL-1β,
AKT1, PTGS2, BCL2, and JUN may be the core targets of gin-
seng for the treatment of DM-induced ED and asthenospermia.
3.6. Results of GO function and KEGG pathway enrichment
analysis
The results of GO functional enrichment analysis of 44 intersect-
ing targets showed that there were 755 BPs, which were mainly
related to response to lipopolysaccharide (LPS), response to
reactive oxygen species, cellular response to lipid, inammatory
response, etc. There are 15 CCs, mainly including membrane raft,
membrane microdomain, caveola, plasma membrane raft, and
nuclear envelope, There were 66 articles in MF, involving pro-
tein homodimerization activity, estrogen response element bind-
ing, nuclear steroid receptor activity, nuclear receptor activity,
ligand-activated transcription factor activity, etc. After selecting
the Top 10 entries respectively, the bar chart was plotted using
the microbiology letter platform (Fig. 5A). KEGG pathway
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Liu et al. • Medicine (2024) 103:34 Medicine
enrichment analysis yielded 139 pathways, mainly AGE-RAGE
signaling pathway in diabetic complications, TNF signaling path-
way, lipid and atherosclerosis, Epstein-Barr virus infection, and so
on. After selecting the Top 20 pathways with a high correlation
with DM-induced ED and asthenospermia in the gure, the bub-
ble diagram was drawn using the microbiotics platform (Fig. 5B).
3.7. Drug-components-disease-target-pathway network
The constructed Network le and attribute Type le were
uploaded to the Cytoscape 3.9.1 platform, and the drug-
components-disease-target-pathway network diagram was
drawn based on its nodes, edges, and other attributes (Fig. 6).
Figure 1. The schematic illustration of the study.
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Table 1
Basic information of ginseng active components.
No. Active components ID OB (%) DL
1 Diop MOL002879 43.59 0.39
2 Stigmasterol MOL000449 43.83 0.76
3 Beta-sitosterol MOL000358 36.91 0.75
4 Inermin MOL003648 65.83 0.54
5 Kaempferol MOL000422 41.88 0.24
6 Aposiopolamine MOL005308 66.65 0.22
7 Deoxyharringtonine MOL005317 39.27 0.81
8 Dianthramine MOL005318 40.45 0.2
9 Arachidonate MOL005320 45.57 0.2
10 Frutinone A MOL005321 65.9 0.34
11 ginsenoside rh2 MOL005344 36.32 0.56
12 Ginsenoside-Rh4-qt MOL005348 31.11 0.78
13 Girinimbine MOL005356 61.22 0.31
14 Panaxadiol MOL005376 33.09 0.79
15 Suchilactone MOL005384 57.52 0.56
16 alexandrin_qt MOL005399 36.91 0.75
17 Fumarine MOL000787 59.26 0.83
DL = drug-like properties, OB = oral bioavailability.
Figure 2. Ginseng as a potential target for the treatment of DM-induced ED and asthenospermia (A) DM targets obtained from the GeneCard and OMIM. (B)
ED targets obtained from the GeneCard and OMIM. (C) Asthenospermia targets obtained from the GeneCard and OMIM. (D) Intersecting targets of DM-induced
ED and Asthenospermia. (E) Venny diagram of intersecting targets of ginseng involvement in DM-induced ED and Asthenospermia. DM = diabetes mellitus, ED
= erectile dysfunction, OMIM = Online Mendelian Inheritance in Man.
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Liu et al. • Medicine (2024) 103:34 Medicine
3.8. Molecular docking validation
The core ginseng components kaempferol, beta-sitosterol, ginse-
noside rh2, stigmasterol, and fumarine were molecularly docked
with the core targets TNF (PDB ID: 2TUN), IL-1β (PDB ID:
1HIB), AKT1 (PDB ID: 1H10), PTGS2 (PDB ID: 5F19), BCL2
(PDB ID: 1G5M), and JUN were subjected to molecular dock-
ing (Table 4), thus validating the results obtained from network
pharmacology. The binding energy in molecular docking was
<0 kcal/mol suggesting spontaneous binding between the core
component (ligand) and the core target (receptor), and when
the binding energy was ≤−5.0 kcal/mol, it suggested good bind-
ing between the docked core component and the core target. As
shown in Table 4, the docking energies were all ≤−5.0 kcal/mol,
indicating that the binding conformation between the core com-
ponents and core target proteins was stable, among which the
core component with the lowest binding energy to AKT1 (PDB
ID: 1H10) and PTGS2 (PDB ID: 5F19) was kaempferol; and
the core components with the lowest binding energies to TNF
(PDB ID: 2TUN), IL-1β (PDB ID: 1HIB), JUN (PDB ID: 1JNM)
is ginsenoside rh2; the lowest binding energy to BCL2 (PDB ID:
1G5M) is fumarine. PyMOL software was used to visualize the
binding conformations of the above 6 core targets with kaemp-
ferol, ginsenoside rh2, and fumarine, respectively (Fig. 7A–F),
where the key active components (ligands) are shown in green,
the target proteins (receptors) are shown in light blue, and the
dotted line in yellow shows the hydrogen bonding effect.
4. Discussion
As common complications of DM, the development of ED and
asthenospermia seriously affects the quality of life of young and
middle-aged men. In recent years, studies have found that TCM
has shown unique advantages in the treatment of DM-induced
ED and asthenospermia. Previous studies have shown that gin-
seng has an ameliorative effect on these 2 diseases,[27,28] but the
specic mechanism of ginseng’s action on ED and asthenosper-
mia induced by the common pathogenic factor – DM has not
been fully elucidated. Therefore, network pharmacology and
molecular docking methods were applied to investigate the
mechanism of ginseng’s “the same treatment for different dis-
eases” in this study.
In this study, a total of 17 active ingredients of ginseng, 255
targets of action, and 44 targets of drug-disease intersection
Figure 3. Drug-active components-intersection target network. The ginseng is shown as orange triangles, the active ingredients are shown as red diamonds,
and the intersecting targets are shown as purple circles.
Table 2
Topological analysis of ginseng core components.
No. Core components Molecule ID DC
1 Kaempferol MOL000422 28
2 Beta-sitosterol MOL000358 18
3 ginsenoside rh2 MOL005344 10
4 Stigmasterol MOL000449 9
5 Fumarine MOL000787 6
DC = degree value.
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Liu et al. • Medicine (2024) 103:34 www.md-journal.com
were screened by network pharmacology. The active compo-
nents with the highest rankings of the degree value of the tar-
get of action were kaempferol, beta-sitosterol, ginsenoside rh2,
stigmasterol, and fumarine. Studies have conrmed that kaemp-
ferol has a variety of pharmacological effects such as antiox-
idant, anti-inammatory, anticancer, and anti-diabetic. Oboh
et al[29] found that kaempferol signicantly inhibited lipid per-
oxidation and scavenging of superoxide anions in the reduced
coenzyme II (NADPH) or Fe2+-induced lipid microsystems, thus
having an ameliorative effect on ED in the penis. Huang et al[30]
found that kaempferol was able to signicantly inhibit LPS-
induced expression of the MAPK pathway in THP-1 of human
monocytes, and reduce the production of inammatory factors
such as macrophage-derived chemokines, interferon-inducible
protein-10, and interleukin-8, thus effectively inhibiting the occur-
rence of inammation. Beta-sitosterol has pharmacological effects
such as anti-inammatory, antioxidant, anti-atherosclerotic,
and hypoglycemic. It has been found that beta-sitosterol can
effectively scavenge oxygen free radicals, reduce the level of
intracellular peroxidase, and play an antioxidant role.[31,32]
Beta-sitosterol can also inhibit macrophage IL-6 activity, reduce
the secretion of inammatory factors such as IL-1, TNF, and
play an anti-inammatory role.[33] It was found that ginseno-
side rh2 possesses pharmacological effects such as hypoglyce-
mic, hypolipidemic, antioxidant, and anti-atherosclerotic. Hou
et al[34] found that ginsenoside rh2 has the effect of increasing
the activity of antioxidant enzymes such as SOD, reducing
ROS, and decreasing oxidative stress. In addition, ginseno-
side rh2 has anti-inammatory activity, which can inhibit the
release of pro-inammatory cytokines and inhibit the inam-
matory signaling pathway, effectively improving the inamma-
tory response.[35] Jie et al[36] found that stigmasterol can play
an anti-inammatory role by regulating the expression of 2
core targets, MAPK3 and PRKACA, on the estrogen signaling
pathway and modulating cytokine levels. Fumarine was found
to signicantly reduce the levels of LPS-induced inammatory
cells (neutrophils and macrophages) as well as the expression of
inammatory factors through inhibition of the Toll-like receptor
4 (TLR4) pathway.[37] It also inhibited the secretion of inducible
nitric oxide synthase and cyclooxygenase-2 (COX-2), thereby
suppressing the production of inammatory factors.[38] The
above results indicate that a variety of active compounds in gin-
seng interact together to exert multiple synergistic effects such
as anti-inammatory, antioxidant, and hypoglycemic effects.
This may be the pharmacological basis of ginseng in the treat-
ment of ED and asthenospermia.
The PPI network analysis showed that the process of gin-
seng treatment for DM-induced ED and asthenospermia mainly
involved targets such as TNF, IL-1β, AKT1, PTGS2, BCL2, and
JUN. These targets are predominantly associated with inamma-
tion, oxidative stress and apoptosis, consistent with the disease
characteristics and pathogenesis of ED and asthenospermia. The
above 6 key targets were characterized and molecular docking
with the active ingredients was carried out. The results showed
that the core components of ginseng bound well to the 6 core
targets, of which TNF (PDB ID: 2TUN) bound most strongly
to ginsenoside rh2. TNF promotes T and B lymphocyte acti-
vation and regulation of inammation-associated acute-phase
responses, and is considered to be a mediator of inamma-
tory and immune responses.[39] IL-1β, known as an important
member of the IL-1 family, is an inammatory cytokine located
upstream in the cytokine cascade. It is widely recognized as a
mediator necessary for the effective initiation of innate immunity
and the development of an adaptive immune response, acting as
an amplier of the immune response.[40] Studies of patients with
diabetes-induced ED have found that diabetes-induced hyper-
glycemia signicantly elevates the release of TNF-α and IL-1β
in the endothelium of the testes and penile corpus cavernosum.
This leads to an increased rate of apoptosis in testicular and
penile corpus cavernosum endothelial cells, which in turn affects
normal penile erectile function and spermatogenesis.[41,42] As one
of the subtypes of AKT, AKT1 is closely related to apoptosis
Figure 4. Screening for the core targets. The intersecting targets are shown as pink circles and the core targets are shown as red circles.
Table 3
Topological analysis of core targets.
No. Core targets BC CC DC
1 TNF 183.73862 0.877551 74
2 IL-1B 140.53221 0.84313726 70
3 AKT1 156.87091 0.8113208 68
4 PTGS2 101.48632 0.8269231 68
5 BCL2 38.607296 0.76785713 62
6 JUN 45.446724 0.76785713 62
BC = betweenness centrality, CC = cell components, DC = degree value.
8
Liu et al. • Medicine (2024) 103:34 Medicine
Figure 5. Enrichment analysis. (A) Top 10 GO function enrichment analysis. The vertical coordinate indicates the number of enriched genes, BP represents
biological processes, CC represents cell components, and MF represents molecular functions. (B) Top 20 KEGG pathway enrichment analysis. The horizontal
coordinates indicate the index of GeneRatio, while the vertical coordinates indicate KEGG enrichment entries. The index of GeneRatio represents the ratio of the
number of pathway-related targets, and it represents the number of annotated targets in certain pathways. The higher the score of GeneRatio, the higher the
level of enrichment. The size of the dots represents the number of targets in their representative pathways. The more the genes involved, the larger the bubble.
The color of the dot presents the different P-values. BP = biological processes, CC = cell components, GO = gene ontology, KEGG = Kyoto Encyclopedia of
Genes and Genomes, MF = molecular functions.
Figure 6. Drug-components-disease-target-pathway network diagram. The drug is shown as blue triangles, components are shown as teal ovals, diseases are
shown as yellow triangles, targets are shown as purple hexagons, and pathways are shown as pink diamonds.
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Liu et al. • Medicine (2024) 103:34 www.md-journal.com
and cell survival, proliferation and metabolism. In DM and its
complications, activated AKT increases oxygen consumption,
leading to the generation of ROS and oxidative stress, which
in turn accelerates a cell death.[43] BCL2 is the major apoptosis-
regulating gene. It has been shown that glucose interacts with
pancreatic β-cells in a prolonged state of high glucose, which
leads to β-cell apoptosis. Their apoptosis is associated with a
balance in the family of b-cell lymphoma (BCL) genes.[44]
The Metascape online tool (https://metascape.org/) was
applied to perform GO function and KEGG pathway enrich-
ment analyses of the intersecting targets. The GO enrichment
analysis revealed that the BP enrichment in ginseng for treating
DM-induced ED and asthenospermia mainly involves response
to LPS, response to reactive oxygen species, cellular response to
lipid, etc. The MF enrichment mainly involved protein homod-
imerization activity, estrogen response element binding and
nuclear steroid receptor activity. Enrichment analysis of KEGG
pathways showed that they are mainly involved in AGE-RAGE
signaling pathway in diabetic complications, TNF signaling
pathway, Lipid and atherosclerosis, etc. The AGE-RAGE sig-
naling pathway in diabetic complications is involved in a wide
range of biological processes in organisms, and AGEs trigger
a range of pathologies mainly through binding to their recep-
tor RAGE. In DM-induced ED and asthenospermia, the for-
mation of AGEs and their interaction with RAGE can lead to
the transcription of inammatory genes and the development
Table 4
Binding affinity of components and receptors.
Target PDB ID Molecule Binding affinity (kcal/mol)
TNF 2TUN Kaempferol −8.5
Beta-sitosterol −6.3
ginsenoside rh2 −9.5
Stigmasterol −6.5
Fumarine −9.0
IL-1B 1HIB Kaempferol −7.2
Beta-sitosterol −6.4
ginsenoside rh2 −7.3
Stigmasterol −7.0
Fumarine −6.7
AKT1 1H10 Kaempferol −6.4
Beta-sitosterol −5.9
ginsenoside rh2 −6.1
Stigmasterol −5.7
Fumarine −6.0
PTGS2 5F19 Kaempferol −9.2
Beta-sitosterol −6.7
ginsenoside rh2 −8.4
Stigmasterol −7.4
Fumarine −9.1
BCL2 1G5M kaempferol −7.8
Beta-sitosterol −5.7
ginsenoside rh2 −7.2
Stigmasterol −5.5
Fumarine −8.6
JUN 1JNM Kaempferol −7.4
Beta-sitosterol −6.8
ginsenoside rh2 −9.3
Stigmasterol −5.9
Fumarine −8.0
Figure 7. Molecular docking results. (A) TNF was docked to ginsenoside rh2. (B) IL-1β was docked to ginsenoside rh2. (C) AKT1 was docked kaempferol. (D)
PTGS2 was docked to kaempferol. (E) BCL2 was docked to fumarine. (F) JUN was docked to ginsenoside rh2.
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Liu et al. • Medicine (2024) 103:34 Medicine
of oxidative stress, which ultimately leads to abnormalities
in penile and testicular structure and function.[45] The TNF
signaling pathway refers to a series of cellular signaling pro-
cesses mediated by TNF. TNF is a bioactive peptide secreted
by activated monocytes or macrophages and is an important
pro-inammatory factor. It activates macrophages, monocytes,
neutrophils, and lymphocytes, which promotes the expression
of inammatory factors such as IL-6 and IL-10, resulting in
exacerbation of the inammatory response.[46] It was found
that plasma concentrations of TNF-α were increased in DM
rats and positively correlated with ED and asthenospermia
conditions.[47,48]
The targets and pathways of ginseng in the treatment of
DM-induced ED and asthenospermia are extremely similar,
which can be regulated through multidimensional networks
of oxidative stress and inammation to achieve the effect of
“treating different diseases with the same treatment.” However,
network pharmacological prediction is a theoretical study and
has certain limitations. First of all, data on bioactive com-
pounds, targets, and molecular interactions from Network
Pharmacology may not comprehensively cover all relevant
interactions or may contain outdated or erroneous informa-
tion, leading to potential bias or inaccuracy in predictions.
Secondly, the complexity of ginseng’s chemical composition
and its interactions with biological systems can pose challenges
in accurately capturing and modeling these interactions in a
network pharmacology framework. Thirdly, Ginseng has been
used in traditional medicine for centuries, and its therapeutic
effects are often attributed to synergistic interactions among
multiple components. Network pharmacology approaches may
not fully capture the holistic and synergistic nature of ginseng’s
pharmacology, particularly if they focus solely on individual
compounds or molecular targets. Fourthly, different species
and cultivars of ginseng may contain varying levels of bioac-
tive compounds and exhibit different pharmacological prop-
erties. Finally, this study may also exhibit some limitations in
terms of experimental validation. In previous studies, animal
models of diabetes-induced ED and diabetes-induced oligozo-
ospermia have been successfully established. However, a model
specically aligned with the focus of this study on diabetes-
induced ED and oligozoospermia has not been developed,
potentially leading to a lack of a degree of accuracy in mecha-
nism validation.
In conclusion, the results of this study provide a new theoreti-
cal basis for the treatment of DM-induced ED and asthenosper-
mia with ginseng on the basis of the TCM theory of “the same
treatment for different diseases.”
Author contributions
Conceptualization: Liming Liu, Xiping Xing.
Data curation: Liming Liu, Yuanfeng Zhang, Jiashu Yang.
Funding acquisition: Xiping Xing.
Software: Liming Liu, Yuanfeng Zhang.
Supervision: Yuanfeng Zhang, Xiping Xing.
Validation: Jiashu Yang, Wenfang Chen, Yibo Shi.
Writing – original draft: Liming Liu, Yuanfeng Zhang, Jiashu
Yang, Xiaogang Zhang.
Writing – review & editing: Liming Liu, Yuanfeng Zhang,
Kaijian Lan, Xiping Xing.
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