Exploring the Mechanism of Yougui Pill against Aging Based on Network Pharmacology and Molecular Docking Study

Abstract

Objective

This study aimed to explore the mechanism of obtaining yang from yin in Yougui pill against aging based on network pharmacology and molecular docking technology.

Methods

The active components and targets of Yougui Pill were obtained by searching the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database and the Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine (BATMEN-TCM) database, and kidney deficiency syndrome-related targets were obtained in the Symptom Mapping (SymMap) Database, a traditional Chinese medicine (TCM) syndrome correlation database. The protein–protein interaction (PPI) network was constructed by using the STRING11.5 database. Then, we used CytoScape3.9.0 software to construct the network of TCM–active components–potential targets, and the core TCM components and core targets of Yougui Pill for the treatment of kidney deficiency were obtained. The function analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis were performed in the Database for Annotation, Visualization, and Integrated Discovery (DAVID). Finally, preliminary verification was performed with the help of molecular docking technology.

Results

A total of 147 active components of 9 drugs of Yougui Pill (Fuzi [Aconiti Lateralis Radix Praeparata], Shudihaung [Rehmanniae Radix Praeparata], Gouqi [Lycii Fructus], Shanyao [Rhizoma Dioscoreae], Shanzhuyu [Corni Fructus], Tusizi [Cuscutae Semen], Danggui [Angelicae Sinensis Radix], Duzhong [Eucommiae Cortex], Lujiaojiao [Cervi Cornus Colla]) were obtained, corresponding to 233 targets. A total of 2,235 targets related to kidney deficiency syndrome and 43 potential therapeutic targets were obtained after the intersection. The core TCM components mainly included quercetin, kaempferol, diosgenin, β-carotene, etc. The core targets involved Trp53 (Tp53), Akt1, Pparg, Nr3c1, App, Casp8, Mapk1, Cav1, and Ctnnb1. A total of 27 biological processes, 10 cellular components, and 11 molecular functions were obtained by gene function enrichment analysis, mainly related to the regulation of gene expression, cell apoptosis and proliferation, and the response to estrogen. A total of 51 KEGG signaling pathways, mainly involving a variety of cancer pathways, apoptosis pathways, longevity regulation pathways, etc.

Conclusion

Yougui Pill can play a role in preventing and treating kidney deficiency syndrome through multiple targets and pathways.

Keywords

Yougui Pill - aging - kidney deficiency syndrome - obtaining Yang from Yin - network pharmacology - molecular docking

Introduction

Aging is an inevitable stage in human life, primarily manifested through gradual morphological changes in the human body and degenerative alterations in cellular, tissue, and organ functions with advancing age. A national survey investigating aging perceptions and the application of traditional Chinese medicine (TCM) antiaging strategies revealed that 95.94% of Chinese residents aged over 18 self-reported exhibiting signs of aging. Notably, the majority of participants identified approximately 30 years old as the optimal age to initiate antiaging interventions. Cross-analysis further indicated that chronic diseases such as hypertension and diabetes mellitus are demonstrating a trend of earlier onset in younger populations.[1]

TCM possesses a profound historical legacy in aging research, encompassing comprehensive theoretical frameworks and clinical. According to TCM philosophy, yin and yang represent interdependent yet opposing forces within a unified system. In human physiology, qi embodies yang characteristics while blood corresponds to yin principles. This dualistic concept permeates all natural phenomena, where seasonal variations and yin–yang interactions constitute the fundamental dynamics governing universal development and human growth. The Yellow Emperor's Inner Classic (Huang Di Nei Jing) emphasizes that adherence to natural yin–yang rhythms through disciplined lifestyle practices enables physiological harmony, thereby promoting health and longevity.

The dynamic interplay of yin and yang finds manifestation through the five elements (metal, wood, water, fire, and earth), with each element demonstrating distinct yin–yang attributes. According to this framework, the kidney system corresponds to the water element and serves as the physiological foundation for yin–yang equilibrium in zang-fu organs (internal organ systems). Specifically, the renal substances that nourish visceral functions constitute kidney yin, which facilitates human reproduction, growth, and development by providing essential biochemical substrates. Conversely, the functional impetus driving organic physiological activities is termed kidney yang, representing the motivating impetus of vital functions. This theoretical construct receives validation in classical texts. Yellow Emperor's Inner Classic (Huang Di Nei Jing) analogizes human yang qi to celestial solar energy, asserting that disruption of yang qi circulation leads to diminished vitality and attenuated life functions. Notably, the eminent physician, Sun Simiao, documented progressive yang deficiency commencing around the age of 50, clinically correlating with aging manifestations. These historical observations substantiate the TCM tenet that “yang deficiency” constitutes the pivotal mechanism in senescence.

The Ming Dynasty physician, Zhang Jinyue (1563–1640), formulated the yin–yang integration doctrine, positing that yin and yang share ontological unity through mutual dependence and transformative interchange. This theoretical framework established the clinical principle of prioritizing yin–yang pattern differentiation to restore dynamic equilibrium. Significantly, Zhang Jinyue developed the seminal therapeutic strategy of securing yang through yin nourishment, epitomized by his creation of Yougui Pill—a quintessential yang-tonifying formula derived from yin-enriching components. Yougui Pill, found in Volume 51 of Jingyue Quanshu, consists of 10 traditional Chinese herbs, including Rougui (Cinnamomi Cortex), Fuzi (Aconiti Lateralis Radix Praeparata), Lujiaojiao (Cervi Cornus Colla) as the primary herbs to warm and tonify kidney Yang, Shudihuang (Rehmanniae Radix Praeparata), Gouqi (Lycii Fructus), Shanyao (Rhizoma Dioscoreae), Shanzhuyu (Corni Fructus) serve as secondary herbs to nourish yin, tonify blood and kidneys, and benefit the liver and spleen, fulfilling the role of obtaining yang from yin. Additionally, Tusizi (Cuscutae Semen), Danggui (Angelicae Sinensis Radix), and Duzhong (Eucommiae Cortex) are used to strengthen tendons and bones, nourish the blood, and replenish essence. The entire formula has the effect of warming and tonifying kidney yang, replenishing essence and blood, thus Yougui Pill can play a role in slowing down aging by replenishing yang energy in the kidney.

Network pharmacology, grounded in computer technology, network biology, and other related concepts, constructs an interactive network of drug–active ingredient–gene target–disease to systematically and comprehensively observe the intervention and impact of drugs on the disease network. This approach facilitates drug development and the treatment of complex diseases.[2] TCM is characterized by its holistic concept and syndrome differentiation and treatment, where the treatment of diseases involves the synergistic effects of multiple components, pathways, and targets.[3] Therefore, network pharmacology offers a novel approach for the in-depth study of TCM systems and has been widely applied in research on TCM and disease treatment. The aim of this study is to explore the mechanism of action of Yougui Pill in antiaging based on network pharmacology and molecular docking techniques. The flowchart of network pharmacology research ideas is shown in [Fig. 1].

Fig. 1 Flowchart of Yougui Pill in treating kidney deficiency syndrome. DL, drug-likeness; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; OB, oral bioavailability.

Materials

Active Ingredients and Targets of Yougui Pill Screening

The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP, https://tcmsp-e.com/tcmsp.php) and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine (BATMEN-TCM, https://bionet.ncpsb.org.cn/batman-tcm/) databases were used to screen the active ingredients and targets of action of Yougui Pill, and TCMSP not only constructed a drug–target–disease network but also involves the pharmacokinetic properties of natural compounds such as oral bioavailability (OB), drug-likeness (DL), intestinal epithelial permeability, etc. OB ≥30% and DL ≥0.18 were used as the screening conditions to obtain the active ingredients and protein targets of botanicals in Yougui Pill because Rougui (Cinnamomi Cortex) did not meet the above conditions so it was not included in the analysis. In the BATMEN-TCM database, we searched the animal drug Lujiaojiao (Cervi Cornus Colla) to obtain its active ingredient and target. In the Uniprot database (https://www.uniprot.org/), “Status” was set to “Reviewed (Swiss-Prot)” and “Organisms” was set to “Mouse,” and Gene Symbol conversion and correction were performed for the above targets.

Screening of Kidney Deficiency Syndrome Targets

In the “Syndrome” search box of the Symptom Mapping Database, we searched for kidney-related syndromes only. Among them, “Kidney-Yang deficiency syndrome” “Kidney-Yang deficiency syndrome” “Kidney-Yang deficiency and decline syndrome” and “Kidney-Yang deficiency syndrome” contained the same TCM symptoms. Therefore, “Kidney deficiency syndrome” and “Kidney-yang deficiency syndrome” were selected for subsequent analysis. The symptoms corresponding to the above two syndromes were combined, duplicates were removed, and the genes corresponding to each symptom were organized and de-emphasized again in Excel 2021 to obtain the relevant targets for the kidney deficiency syndrome.

Construction of Traditional Chinese Medicine–Active Ingredient–Potential Target Network

The intersection of active ingredient-related targets and syndrome-related targets of Yougui Pill was taken as the potential targets of Yougui Pill for treating renal deficiency and visualized by Venny 2.1.0, and the network diagram of “TCM–active ingredients–potential targets” was constructed by using CytoScape 3.9.0 software.

Protein–Protein Interaction Protein (PPI) Network Construction, and Core Target Screening

The obtained potential target genes were imported into the STRING11.5 database (https://cn.string-db.org/), and the species was set as “Mus musculus,” the minimum interaction threshold was set as “highest confidence” ≥0.4 and hid the free nodes, exported the data, visualized the PPI network with CytoScape 3.7.0 software, and used CytoNCA plug-in to perform network topology analysis, and screened the core targets with betweenness centrality, closeness centrality, and degree centrality ≥median of all nodes.

Bioinformatics Enrichment Analysis of Core Target Genes

Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment were performed on the screened core target genes using the DAVID database (https://david.ncifcrf.gov/summary.jsp). The GO enrichment analysis included biological process (BP), cellular component (CC), and molecular function (MF), and the species “Mus musculus” was selected to obtain the BP and metabolic pathway data of “Kidney deficiency syndrome” treated by Yougui Pill, outcome data for which false discovery rate (FDR) ≤0.05 and P < 0.05 were met and were visualized using the online data analysis and visualization web site of Microbiology Letters, and the results are represented as bubble plots.

Molecular Docking

The core targets related to aging were selected for molecular docking with their corresponding active ingredients of Yougui Pill. First, the 2D structure of the active ingredient was searched in the TCMSP database and saved in “MOL2” format, the core target protein information was searched in the Uniprot database, and the 3D structure of the appropriate protein was selected and downloaded as “PDB” format file. The water molecules and original ligands were removed from the protein using PyMOL software. Then, we performed docking in AutoDockTools-1.5.7 and calculated the binding energies between the active ingredient and the core target proteins, and the results were saved in the format of “pdbqt.” The results were saved in “pdbqt” format and finally visualized in PyMOL.

Results

Screening Results of Active Ingredients and Targets of Yougui Pill

A total of 147 active compounds were obtained from Yougui Pill, including 2 Danggui (Angelicae Sinensis Radix), 28 Duzhong (Eucommiae Cortex), 21 Fuzi (Aconiti Lateralis Radix Praeparata), 45 Gouqi (Lycii Fructus), 16 Shanyao (Rhizoma Dioscoreae), 20 Shanzhuyu (Corni Fructus), 2 Shudihuang (Rehmanniae Radix Praeparata), 11 Tusizi (Cuscutae Semen), and 2 Lujiaojiao (Cervi Cornus Colla). The results are shown in [Table 1] (listing the top five compounds for each herb based on OB value). A total of 233 active ingredient targets were obtained from Yougui Pill (after weight removal).

Table 1 Active ingredients of Yougui Pill in traditional Chinese medicine

Source	Mol ID	Molecule name	OB / %	DL
Duzhong (Eucommiae Cortex)	MOL006709	AIDS214634	92.43	0.55
	MOL009009	(+)-medioresinol	87.19	0.62
	MOL009042	Helenalin	77.01	0.19
	MOL009031	Cinchonan-9-al,6′-methoxy-, (9R)-	68.22	0.40
	MOL004367	Olivil	62.23	0.41
Fuzi (Aconiti Lateralis Radix Praeparata)	MOL002421	Ignavine	84.08	0.25
	MOL002419	(R)-Norcoclaurine	82.54	0.21
	MOL002398	Karanjin	69.56	0.34
	MOL002388	Delphin_qt	57.76	0.28
	MOL002395	Deoxyandrographolide	56.30	0.310
Shanyao (Rhizoma Dioscoreae)	MOL000546	Diosgenin	80.88	0.81
	MOL005429	Hancinol	64.01	0.37
	MOL000310	Denudatin B	61.47	0.38
	MOL001736	(−)-taxifolin	60.51	0.27
	MOL005430	Hancinone C	59.05	0.39
Shanzhuyu (Corni Fructus)	MOL005531	Telocinobufagin	69.99	0.79
	MOL005552	Gemin D	68.83	0.56
	MOL005360	Malkangunin	57.71	0.63
	MOL005486	3,4-Dehydrolycopen-16-al	46.64	0.49
	MOL001495	Ethyl linolenate	46.10	0.20
Gouqi (Lycii Fructus)	MOL009664	Physalin A	91.71	0.27
	MOL008400	Glycitein	50.48	0.24
	MOL006209	Cyanin	47.42	0.76
	MOL009651	Cryptoxanthin monoepoxide	46.95	0.56
Tusizi (Cuscutae Semen)	MOL005944	Matrine	63.77	0.25
	MOL001558	Sesamin	56.55	0.83
	MOL006649	Sophranol	55.42	0.28
	MOL000354	Isorhamnetin	49.60	0.31
Gouqi (Lycii Fructus)/Tusizi (Cuscutae Semen)	MOL000098	Quercetin	46.43	0.28
Danggui (Angelicae Sinensis Radix)/Shudihuang (Rehmanniae Radix Praeparata)	MOL000449	Stigmasterol	43.83	0.76
Danggui (Angelicae Sinensis Radix)	MOL000358	β-sitosterol	36.91	0.75
Shudihuang (Rehmanniae Radix Praeparata)	MOL000359	Sitosterol	36.91	0.75
Lujiaojiao (Cervi Cornus Colla)	–	Calcium phosphate	–
		Calcium carbonate

Targets Related to Renal Deficiency

A total of 112 symptoms related to the kidney deficiency syndrome were obtained, with a total of 2,235 related targets, and 43 potential therapeutic targets were obtained by taking the intersection with the targets of the active ingredient of Yougui Pill, as shown in [Fig. 2].

Fig. 2 Targets of Yougui Pill for treating kidney deficiency syndrome.

“Traditional Chinese Medicine–Active Ingredient–Potential Target” Network Analysis

As shown in [Fig. 3], in the network diagram of “TCM–active ingredient–target,” the top eight core active ingredients of TCM are shown in [Table 2], which may play a key role in the treatment of renal deficiency in Yougui Pill.

Fig. 3 Traditional Chinese medicine–active ingredient–potential target network diagram.Notes: Triangles are traditional Chinese herbs, hexagons are active ingredients of traditional Chinese herbs, and rhombuses are potential targets for the treatment of kidney deficiency by Yougui Pill.

Table 2 Core Chinese medicine active ingredients

Mol ID	Molecule name	Degree
MOL000098	Quercetin	27
MOL000422	Kaempferol	9
MOL000546	Diosgenin	8
MOL002773	β-carotene	7
MOL000358	β-sitosterol	6
MOL009015	(−)-Tabernemontanine	6
MOL001558	Sesamin	5
MOL000449	Stigmasterol	5

Construction and Analysis of Protein–Protein Interaction Network

As shown in [Fig. 4], there were 43 nodes and 202 edges in the PPI network graph of potential therapeutic targets, of which Ier3ip1, Slc12a6, Slc12a1, and Adra2b were unassociated free nodes, and therefore were not included in the subsequent analysis. Network topology analysis was performed in CytoScape 3.9.0 software using centiscape2.2 plug-in and CytoNCA plug-in for three parameters, and nine core targets were obtained with betweenness centrality ≥36.76923077, closeness centrality ≥0.013971292, and degree centrality ≥10.35897436 as the screening criteria, which were Trp53 (Tp53), Akt1, Pparg, Nr3c1, App, Casp8, Mapk1, Cav1 and Ctnnb1, as shown in [Table 3].

Fig. 4 Potential target PPI network.Notes: The node color and size were adjusted according to the degree value, the larger the graph is, the larger the degree value is. PPI, protein–protein interaction.

Table 3 The degree centrality, betweenness centrality, and closeness centrality of core targets

Target spot	Degree centrality	Betweenness centrality	Closeness centrality
Akt1	28	229.0662005	0.020408163
Trp53	27	234.1421523	0.020000000
Ctnnb1	23	166.5397935	0.018181818
Casp8	22	100.3411311	0.017543860
Pparg	20	70.36731047	0.017241379
Mapk1	18	83.13417138	0.016393443
Cav1	16	71.30497280	0.016393443
Nr3c1	14	60.61864247	0.015151515
App	14	105.0350871	0.015151515

Abbreviations: BC, betweenness centrality; CC, closeness centrality; DC, degree centrality.

Enrichment Analysis

Gene Ontology Analysis Results

Among the results obtained from the analysis of the DAVID database, a total of 48 entries satisfied an FDR of ≤0.05 and P < 0.05, of which 27 were BP, 10 were CC, and 11 were MF. The BP mainly includes regulation of gene expression, positive regulation of apoptotic process, response to estrogen, cardiac development, positive regulation of apoptotic process in neurons, RNA polymerase II promoter transcription regulation, etc. The top 20 were taken to draw the visualization bubble diagram. The CC mainly includes macromolecular complexes, mitochondria, cytosol, cytoplasm, nucleoplasm, RNA polymerase II transcription factor complexes, etc. The MF is dominated by enzyme-binding, protein-binding, chromatin-binding, and protein kinase-binding, etc., as shown in [Fig. 5].

Fig. 5 GO enrichment analysis. GO, Gene Ontology. Notes: A, BP; B, CC; C, MF.

Kyoto Encyclopedia of Genes and Genomes Signaling Pathway Analysis Results

A total of 51 pathways had an FDR of ≤0.05 and P < 0.05, mainly involving pathways of multiple cancers, lipid and atherosclerosis pathways, thyroid hormone signaling pathways, apoptosis pathways, fluid shear stress and atherosclerosis, longevity regulation pathways, and endocrine resistance pathways, etc. the top 20 pathways with the most significance were visualized, as shown in [Fig. 6].

Fig. 6 KEGG pathway enrichment analysis. KEGG, Kyoto Encyclopedia of Genes and Genomes.

Molecular Docking Results

The results of core target enrichment analysis showed that a total of three targets, Tp53, Akt1, and Pparg, were enriched in the longevity regulation pathway, so molecular docking was performed with the three target proteins mentioned above as the receptor, and the molecular docking was performed with their corresponding active ingredients, and the results of docking are shown in [Table 4]. It is generally believed that the smaller the binding energy between the active ingredient and the target protein, the stronger the binding effect, with a binding energy of < −5.0 kcal/mol indicating a better binding performance, and < −7 kcal/mol indicating a stronger binding performance.[4] The active ingredient of Yougui Pill with a binding energy of < −5.0 kcal/mol and hydrogen bonding was selected for visualization with the target protein, and the results are shown in [Fig. 7].

Table 4 Molecular docking results

Target	Mol ID active ingredient		Combined energy (kcal/mol)
Tp53	MOL000546	Diosgenin	−7.40
	MOL000098	Quercetin	−5.32
Akt1	MOL000546	Diosgenin	−7.52
	MOL002773	Beta-carotene	−6.21
	MOL000422	Kaempferol	−6.02
	MOL000098	Quercetin	−6.02
Pparg	MOL008457	Tetrahydroalstonine	−4.84
	MOL008240	(E)-3-[4-[(1R,2R)-2-hydroxy-2-(4-hydroxy-3-methoxy-phenyl)-1-methylol-ethoxy]-3-methoxy-phenyl]acrolein	−0.47
	MOL000422	Kaempferol	−2.79
	MOL000098	Quercetin	−2.68
	MOL011604	Syringetin	−2.68
	MOL008400	Glycitein	−3.41
	MOL000354	Isorhamnetin	−3.15

Fig. 7 Molecular docking model.Notes: (A) schematic diagram of Akt1 and quercetin docking. (B) schematic diagram of Akt1 and kaempferol docking. (C) schematic diagram of Akt1 and Diosgenin docking. (D) schematic diagram of TP53 and Diosgenin docking.

Discussions

In this study, through the analysis of the TCM–active ingredient–potential target network, it was found that active ingredients in Yougui Pill, such as quercetin, kaempferol, and diosgenin, may play crucial roles in the treatment of kidney deficiency syndrome. Flavonoids, abundant natural products in plants and vegetables, possess antioxidant, anti-inflammatory, and anticancer properties, making them effective in treating various age-related diseases such as atherosclerosis, Alzheimer's disease, and cancer. Both quercetin and kaempferol belong to the flavonoid family.[5] In terms of bone metabolism regulation, quercetin can modulate bone marrow mesenchymal stem cells through estrogen signaling and MAPK pathways, promoting bone formation and inhibiting bone resorption, thereby effectively improving osteoporosis.[6] In improving the reproductive system, quercetin maintains the dynamic balance of maternal–fetal immunity, which is beneficial for the prevention and treatment of adverse pregnancy outcomes such as recurrent miscarriage and infertility.[7] Experiments have also shown that high doses of quercetin can increase serum progesterone levels in rats, reduce the apoptosis rate of ovarian granulosa cells, and protect the reproductive system.[8] In preventing brain aging, quercetin improves learning ability by protecting neurons, reducing oxidative damage, and inhibiting inflammation, playing a role in the prevention and treatment of neurodegenerative diseases.[9] Furthermore, studies have indicated that quercetin can increase the expression of genes such as brain-derived neurotrophic factor and nerve growth factor, and accelerate neurogenesis in adult rats, thereby alleviating symptoms associated with Alzheimer's disease.[10] As a high-polarity glycoside, kaempferol has poor absorption, but its combination with quercetin can enhance its bioavailability and thus its biological effects. High intake can effectively prevent and treat various cancers such as cervical cancer, renal cell carcinoma, bladder cancer, and prostate cancer.[11] In bone metabolism regulation, kaempferol can enhance the viability of C28/I2 inflammatory chondrocytes induced by lipopolysaccharide and inhibit apoptosis.[12] In improving the reproductive system, kaempferol exhibits progesterone activity and can serve as a plant progesterone to promote female reproductive health.[13] In preventing brain aging, kaempferol protects cranial nerves and improves cognitive ability, mainly through mechanisms involving the scavenging of free radicals, enhancing neuronal antioxidant capacity, and regulating neurotransmitter levels. Studies have found that metabolic changes in the cortical region near the striatum and the hippocampus are associated with cognitive dysfunction, visuospatial deficits, memory decline, and difficulties in learning new skills. High neurotoxic A1 astrocytes and β-amyloid peptides have specific correlations with brain degenerative diseases, and kaempferol can effectively prevent the generation of reactive A1 astrocytes and the activation of their marker complement C3 protein in the striatum and hippocampus, as well as inhibit the increase of β-amyloid peptides, thereby protecting against neurodegenerative diseases.[14] Diosgenin exhibits neuroprotective, anticancer, and antiatherosclerotic effects, and is also used to treat male erectile dysfunction, enhance cognitive function, and alleviate menopausal symptoms in women.[15] In terms of antiaging, studies have shown that diosgenin exerts antiaging effects by inhibiting the phosphoinositide 3-kinase(PI3K) - protein kinase B(PKB, also known as AKT)-mechanistic target of rapamycin kinase(mTOR) signaling pathway and reducing reactive oxygen species production.[16] Diosgenin can reduce fas-dependent and mitochondria-dependent apoptotic pathways in neurons of aging rats induced by D-galactose and enhance survival pathways related to the Bcl-2 family of neurons and the Insulin-like Growth Factor 1 (IGF-1)-PI3K-AKT pathway, thereby preventing neuronal apoptosis, promoting neuronal survival, and preventing brain aging.[17] The number of oocytes in a woman's ovaries decreases with age and is not renewable. After the age of 30, a woman's ovarian reserve significantly declines, while diosgenin can increase the number of primordial follicles, improve ovarian reserve in aging mice, and delay ovarian aging.[18]

Through PPI protein network analysis, nine core targets were screened, among which Tp53, Akt1, and Pparg were enriched in the longevity regulation pathway. Analysis of two mouse genomes revealed that TP53 is a DNA repair and DNA damage signaling gene that plays a role in the maintenance of genomes of long-lived species.[19] Moreover, as a tumor suppressor gene, mutations in Tp53 have been detected in many cancers.[20] Additionally, its downstream target gene TIGAR can treat cardiovascular diseases such as atherosclerosis by inhibiting glycolysis and reducing oxidative stress.[21] Akt1/2 knockout mice exhibit premature muscle loss, systemic insulin resistance, reduced bone mass, and shortened lifespan, indicating that skeletal muscle Akt plays a crucial role in regulating skeletal muscle aging and lifespan.[22] The PI3K/AKT/mTOR signaling pathway regulates neuronal apoptosis, autophagy, neuronal cell proliferation, and increased synaptic plasticity. Abnormal activation of this pathway can affect these physiological functions and even trigger neurological diseases.[23] The PI3K/Akt signaling pathway is also involved in the growth and development of testicular tissue and is related to androgen secretion.[24] Pparg controls the differentiation of osteoblasts and adipocytes, and increased Pparg activity can lead to bone turnover and have negative effects on the skeleton.[25] The deficiency of Pparg can also lead to renal insufficiency, manifesting as increased glucosuria and albuminuria.[26] Studies have shown that the expression of Pparg can affect the human immune system,[27] and there are also studies indicating that Pparg is related to aging-associated diseases such as diabetes, obesity, and atherosclerosis.[28] [29] [30]

The results of GO analysis of core targets related to the treatment of renal deficiency syndrome by Yougui Pill showed that Yougui Pill could exert its therapeutic effects through a variety of BPs such as the regulation of gene expression, cell proliferation, and apoptosis, the response to estrogen, the development of the nervous system, and the response to oxidative stress, etc. The results of the analysis of the KEGG signaling pathway showed that multiple signaling pathways such as various cancer pathways, lipid and atherosclerosis, apoptosis, longevity regulation pathways, and neurodegeneration–multiple disorders, etc., were significantly enriched, and all of these BPs and signaling pathways are closely linked with the phenotypes of renal deficiency syndrome.

Conclusion

In summary, this study investigated the active ingredients and action targets of the TCM Yougui Pill for the treatment of renal deficiency syndrome through network pharmacology and used molecular docking technology to preliminarily validate some of the core targets and their corresponding active ingredients, and the results showed that Yougui Pill can play a role in preventing and treating renal deficiency syndrome through multitargets and multiple pathways, but further validation through experiments is still required.

Conflict of Interest

The authors declare that there is no conflict of interest.

CRediT Authorship Contribution Statement

Haiyan Yang: Conceptualization, data curation, funding acquisition, project administration, resources, supervision, and writing—original draft. Qian Shi: Formal analysis, investigation, methodology, validation, and writing—original draft. Chunchun Ji: Funding acquisition, methodology, project administration, software, and writing—review and editing.

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Publication History

Received: 21 November 2024

Accepted: 29 January 2025

Article published online:
08 April 2025