Mechanistic Insights into the Therapeutic Effects of Tiaozang Shumi Mixture on Chronic Constipation

Objective To analyze the roles and mechanisms of lactitol and Bifidobacterium infantis in the treatment of rat constipation and to investigate their effects on aquaporin3 (AQP3) and interstitial cells of Cajal (ICC) in colon tissues. Methods Thirty SD male rats were recruited in this study, 6 of which were randomly selected as the control and the rest were given 4 mg/kg.d of loperamide for 5 consecutive days to construct the rat model of constipation. The rats with constipation were randomly divided into four groups including model group, lactitol treatment group, Bifidobacterium infantis treatment group and lactitol in combination with Bifidobacterium infantis treatment group. General indexes including food intake, water intake, body weight, fecal water content and intestinal transit rate of each rat were measured after receiving corresponding treatments for 7 consecutive days. The levels of substance P (SP) and vasoactive intestinal peptide (VIP) in serums samples were detected by ELISA. The expression of protein kinase A (PKA) and neurokinin-1 receptor (NK-1) at mRNA level in colon tissues were detected by real-time polymerase chain reaction (real-time PCR). Western blot assay and real-time PCR analysis were used to detect the expression of AQP3 and c-kit at protein and mRNA levels, respectively. Results Compared with the rats in model group, the levels of fecal water content and intestinal transit rate, the concentrations of SP and VIP in serums samples, the expression of PKA and NK-1 at mRNA level and the expression of AQP3 and c-kit at mRNA and protein levels were significantly increased in rats from the three treatment groups (P<0.05). The most effective treatment was lactitol in combination with Bifidobacterium infantis, followed by the lactitol treatment and then the Bifidobacterium infantis treatment. Conclusion The combination therapy with lactitol and Bifidobacterium infantis increased the serum levels of SP and VIP in rats with constipation. SP could enhance the contraction of gastrointestinal smooth muscles and improve the intestinal motility by binding to the NK-1 receptor on the membrane of ICC. VIP could promote the absorption of water in intestinal tracts, soften stools and alleviate constipation by upregulating the expression of AQP3 at both protein and mRNA levels via the cyclic adenosine monophosphate-PKA (cAMP-PKA) signaling pathway. Key words: Constipation; Lactitol; Bifidobacterium infantis; Aquaporin3; Interstitial cells of Cajal

The bioactive components of Astragalus membranaceus and Salvia miltiorrhiza improved cardiac and renal function in chronic heart failure (CHF) and chronic kidney disease (CKD), respectively. However, the common regulating molecular mechanisms remain unclear. The aim of this study was to investigate these mechanisms using bioinformatics, network topology, and molecular dynamics simulation techniques. The active components and target sites of A. membranaceus and S. miltiorrhiza were obtained from the Traditional Chinese Medicine Systems Pharmacology database. The targets of CKD and CHF were obtained from various databases for a protein-protein interaction analysis. The Gene Ontology (GO) function and Kyotoencyclopedia of genes and genomes (KEGG) pathway enrichment of intersection targets were analyzed by using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) database. Molecular docking and dynamic simulations were conducted on the core ingredients and targets. The diagnostic efficiency of the key targets was evaluated by using receiver-operating characteristic (ROC) curves. A total of 70 active ingredients and 158 common targets were found. The top five core targets were AKT1, STAT3, TP53, MAPK1, and RELA. The GO enrichment analysis included apoptosis and oxidative stress. The KEGG pathway enrichment results indicated that the drug pair regulated the AGE-receptor for AGE signaling pathway, fluid shear stress and atherosclerosis, and the IL-17 signaling pathway. Molecular docking and dynamic simulations confirmed that the core ingredients had good affinity and stability with the key targets. The ROC curves confirmed the accuracy of every key target for identifying CKD and CHF and demonstrated that combining them improves diagnosis. The combination of A. membranaceus and S. miltiorrhiza proved effective for the treatment of CKD and CHF through various components, targets, and mechanisms. Moreover, it may predict the diagnostic value of key targets, providing a reference for clinical diagnostic applications.

ObjectiveTo explore the effective components and mechanism of Polygonati Rhizoma (PR) in the treatment of osteoporosis (OP) based on network pharmacology and molecular docking methods.MethodsThe effective components and predicted targets of PR were obtained through the Traditional Chinese Medicine Systems Pharmacology and Analysis Platform (TCMSP) database. The disease database was used to screen the disease targets of OP. The obtained key targets were uploaded to the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database for protein-protein interaction (PPI) network analysis. The Database for Annotation, Visualization, and Integrated Discovery (DAVID) was used for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of key targets. Analysis and docking verification of chemical effective drug components and key targets were performed with IGEMDOCK software.ResultsA total of 12 chemically active components, 84 drug target proteins and 84 common targets related to drugs and OP were obtained. Key targets such as JUN, TP53, AKT1, ESR1, MAPK14, AR and CASP3 were identified through PPI network analysis. The results of enrichment analysis showed that the potential core drug components regulate the HIF-1 signaling pathway, PI3K-Akt signaling pathway, estrogen signaling pathway and other pathways by intervening in biological processes such as cell proliferation and apoptosis and estrogen response regulation, with an anti-OP pharmacological role. The results of molecular docking showed that the key targets in the regulatory network have high binding activity to related active components.ConclusionsPR may regulate OP by regulating core target genes, such as JUN, TP53, AKT1, ESR1, AR and CASP3, and acting on multiple key pathways, such as the HIF-1 signaling pathway, PI3K-Akt signaling pathway, and estrogen signaling pathway.

The syndrome of inappropriate antidiuretic hormone secretion is characterized by excessive water uptake and hyponatremia. The extent of hyponatremia, however, is less than anticipated, which is ascribed to a defense mechanism, the vasopressin-escape, and is suggested to involve a tonicity-determined down-regulation of the water channel aquaporin-2 (AQP2). The underlying mechanism, however, is poorly understood. To study this, we used the mouse cortical collecting duct (mpkCCD) cell line. MpkCCD cells, transfected with an AQP2-promoter luciferase construct showed a reduced and increased AQP2 abundance and transcription following culture in hypotonic and hypertonic medium, respectively. This depended on tonicity rather than osmolality and occurred independently of the vasopressin analog dDAVP, cAMP levels, or protein kinase A activity. Although prostaglandins and nitric oxide reduced AQP2 abundance, inhibition of their synthesis did not influence tonicity-induced AQP2 transcription. Also, cells in which the cAMP or tonicity-responsive element (CRE/TonE) in the AQP2-promoter were mutated showed a similar response to hypotonicity. Instead, the tonicity-responsive elements were pin-pointed to nucleotides -283 to -252 and -157 to -126 bp. In conclusion, our data indicate that hypotonicity reduces AQP2 abundance and transcription, which occurs independently of vasopressin, cAMP, and the known TonE and CRE in the AQP2-promoter. Increased prostaglandin and nitric oxide, as found in vivo, may contribute to reduced AQP2 in vasopressin-escape, but do not mediate the effect of hypotonicity on AQP2 transcription. Our data suggest that two novel segments (-283 to -252 and -157 to -126 bp) in the AQP2-promoter mediate the hypotonicity-induced AQP2 down-regulation during vasopressin-escape.

Exploring the potential mechanism of radix astragali against ischemic stroke based on network pharmacology and molecular docking

Objective: To explore the effect of Xinfeng Capsule (XFC) on hypercoagulable state in patients with rheumatoid arthritis (RA) using data mining and network pharmacology. Methods: The data were collected of 524 inpatients with RA who were treated with XFC in the Department of Rheumatology and Immunology of the First Affiliated Hospital of Anhui University of traditional Chinese medicine (TCM) before October 2021. The changes of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), complement component 3 (C3), C4, platelet (PLT), fibrinogen (FBG), thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (APTT) were observed before and after the treatment. By implementing the Apriori module, the association rules between XFC and immune-inflammation indexes and coagulation indexes were analyzed. XFC and disease targets were obtained through traditional chinese medicine systems pharmacology database and analysis platform, Genecards, OMIM, and other databases. The cross targets and core targets were screened, and the network diagram of TCM—active ingredients—potential targets was constructed using Cytoscape3.7.2 software. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed through Database for Annotation, Visualization and Integrated Discovery (DAVID) database. AutoDock Vina software was used for molecular docking between active ingredients and core targets. The docking results were visualized using PyMOL2.3.0 software. Results: (1) Data mining results showed that the inflammation and coagulation indexes of RA patients were significantly improved after XFC treatment, and there was a strong correlation between XFC and the improvement of CRP, ESR, RF, C3, C4, PLT, FBG, TT, PT, and APTT. (2) Network pharmacology results showed that prostaglandin-endoperoxide synthase 2 (PTGS2), CASP3, tumor necrosis factor (TNF), AKT1, and JUN, the main targets of XFC in the treatment of RA, were closely related to apoptosis and were mainly involved in interleukin 17 (IL-17), TNF, and nuclear factor-κB (NF-κb), and other apoptotic and inflammatory signaling pathways. (3) Molecular docking results showed that the active components of XFC, β- sitosterol, and stigmasterol, had good docking with TNF and PTGS2, which might be the key active components of XFC in the treatment of RA-related hypercoagulable state. Conclusion: XFC can improve the hypercoagulable state of patients with RA by promoting cell apoptosis and improving immune inflammatory response.

Non-alcoholic steatohepatitis (NASH) is a predominant metabolic liver disease, typically characterized by hepatic steatosis, oxidative stress, and inflammation. The traditional Chinese medicine Cornus officinalis possesses anti-inflammatory and hepatoprotective pharmacological properties and has shown ameliorative effects on NASH. however, its mechanism of action remains unclear. This study aims to elucidate the mechanisms by which C. officinalis ameliorates NASH. The active components of C. officinalis were analyzed using the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP), and the corresponding targets were predicted. Subsequently, the DisGeNET, GeneCards, and GEO databases were employed to identify NASH-related targets. Venn diagrams were used to intersect the C. officinalis targets with the NASH targets. Protein–protein interaction (PPI) networks were constructed using the STRING database, and PPI network analysis was performed using Cytoscape. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted using the Database for Annotation, Visualization, and Integrated Discovery (DAVID), followed by molecular docking validation. Cornus officinalis was found to contain 20 major active ingredients corresponding to 672 potential targets, 61 of which overlapped with NASH targets. PPI network, GO, and KEGG pathway analyses identified four targets with the highest correlation, and molecular docking results indicated that the active ingredients of C. officinalis exhibited strong binding affinities to NASH targets. The treatment of NASH with C. officinalis is characterized by multiple active ingredients and multiple targets, underscoring the major advantage of traditional Chinese medicine in treating NASH.

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

To screen the key Chinese Herbal Medicines (KCHMs) against breast cancer by data mining, and analyze the potential mechanism of KCHMs using network pharmacology method. Clinical prescriptions consisted of CHMs for treating breast cancer were screened, and then Traditional Chinese Medicine Inheritance Support System (TCMISS) was applied to obtain the KCHMs. Subsequently, active ingredients and corresponding target genes of KCHMs were searched by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database, and target genes of breast cancer were collected using OMIM and MalaCards. After that, the overlapping target genes of KCHMs and breast cancer were screened, and the protein-protein interaction (PPI) network was built. In addition, a network of "KCHMs-active ingredients-breast cancer-targets" was constructed by Cytoscape 3.7.1. Finally, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis were performed with Database for Annotation, Visualization and Integrated Discovery (DAVID) database to reveal the action mechanism of KCHMs. A total of 7 KCHMs were identified, whose active ingredients include quercetin, luteolin, nobiletin, kaempferol, isorhamnetin, naringenin, and be-ta-sitosterol, etc. Based on protein-protein interaction analysis, core targets were ESR1, MYC, CCND1, EGFR, CASP3, ERBB2, etc. Several KEGG pathways (e.g, PI3K-Akt, p53, ErbB, and HIF-1 signaling pathways) were found. Based on the combination of the data mining method and network pharmacology approach, the therapeutic effect of KCHMs on breast cancer may be realized by acting on target genes and signaling pathways related to the formation and progression of breast cancer.

Introduction Aquaporins are a family of transmembrane water channels that aid in osmoregulation, a critical function in the proteoglycan-rich nucleus pulposus. Previous reports have demonstrated expression of aquaporins in the intervertebral disc.1,2 Hypoxia, a defining feature of the nucleus pulposus environment has been shown to regulate expression of aquaporin1 (AQP1) and aquaporin 5 (AQP5).3-6 In some cases, this regulation has been shown to occur through HIF-1α and to depend on hypoxia responsive elements (HREs). The goal of this study was to examine if hypoxia and HIF-1α play a role in regulation and function of aquaporin1 (AQP1) and aquaporin5 (AQP5) in nucleus pulposus (NP) cells of the intervertebral disc. Materials and Methods Quantitative reverse transcription-polymerase chain reaction, Western blot, and immunohistochemistry were used to measure AQP1 and AQP5 expression in nucleus pulposus (NP) and annulus fibrosus (AF) cells and tissues. Transfections were used to determine the role of HIF-1α on AQP1 and AQP5 promoter activity in normoxia and hypoxia. The JASPAR database was used to identify two putative hypoxia response elements (HREs) in the promoter of each aquaporin. HIF-1α levels were modulated with treatment by DMOG or shRNAs. Expression of aquaporins in human tissue samples was evaluated using immunohistochemistry and polymerase chain reaction. Results We found that both AQP1 and AQP5 were expressed in tissues and cells of human and rat discs. To determine whether expression of the aquaporins was regulated by hypoxia, we treated NP cells with hypoxia (1% O2) for 8 to 72 hours. Treatment had no effect on promoter activity, mRNA expression, or protein expression of either aquaporin. Using the JASPAR database, we identified two HREs in the promoter of each aquaporin. Mutation of the HREs in either AQP1 or AQO5 had no suppressive effect on the basal activity of the promoters. We then investigated whether AQP1 and AQP5 were regulated in a HIF1α-dependent manner. Accumulation of HIF-1α by DMOG did not affect expression of either aquaporin. However, suppression of HIF-1α by lentiviral delivery of shRNA significantly decreased both mRNA and protein expression of AQP1 and AQP5. In addition, we have found changes in expression of these aquaporins in human tissue samples from patients with varying degrees of intervertebral disc degeneration. Conclusion These results indicate that, under hypoxic conditions, HIF-1α maintains basal expression of both AQP1 and AQP5 in the NP; however, this regulation is independent of HIF binding to the identified HREs in their promoters. Analysis of human tissue samples suggests that aquaporin expression may be linked to health of the intervertebral disc. Disclosure of Interest None declared References Richardson SM, Knowles R, Marples D, Hoyland JA, Mobasheri A. Aquaporin expression in the human intervertebral disc. J Mol Histol 2008;39(3):303–309 Gajghate S, Hiyama A, Shah M, et al. Osmolarity and intracellular calcium regulate aquaporin2 expression through TonEBP in nucleus pulposus cells of the intervertebral disc. J Bone Miner Res 2009;24(6):992–1001 Tanaka A, Sakurai K, Kaneko K, et al. The role of the hypoxia-inducible factor 1 binding site in the induction of aquaporin-1 mRNA expression by hypoxia. DNA Cell Biol 2011;30(8):539–544 Tie L, Lu N, Pan XY, et al. Hypoxia-induced up-regulation of aquaporin-1 protein in prostate cancer cells in a p38-dependent manner. Cell Physiol Biochem 2012;29(1-2):269–280 Zhang J, Xiong Y, Lu LX, et al. AQP1 expression alterations affect morphology and water transport in Schwann cells and hypoxia-induced up-regulation of AQP1 occurs in a HIF-1α-dependent manner. Neuroscience 2013;252(252):68–79 Kawedia JD, Yang F, Sartor MA, Gozal D, Czyzyk-Krzeska M, Menon AG. Hypoxia and hypoxia mimetics decrease aquaporin 5 (AQP5) expression through both hypoxia inducible factor-1α and proteasome-mediated pathways. PLoS ONE 2013;8(3):e57541

Based on comprehensive network-pharmacology and molecular docking analysis, this study was intended to unveil the multiple mechanisms of Si-Ni-San (SNS) in treating anxious insomnia. The compounds of SNS were meticulously analyzed, selected and standardized with references to their pharmacological attributes. The components included chaihu (Bupleurum chinense DC.), baishao (Paeonia lactiflora Pall.), zhishi (Citrus aurantium L.) and gancao (Glycyrrhiza uralensis Fisch. ex DC.). We used the Traditional Chinese Medicine System Pharmacology (TCMSP) Database, Traditional Chinese Medicines Integrated Database (TCMID), GeneCards database, therapeutic target database (TTD) and comparative toxicogenomic database (CTD) to construct the components-compounds-targets networks and used Cytoscape 3.9.1 software to visualize the outcome. Afterwards, the STRING database and Cytoscape 3.9.1 software were utilized to construct and visualize the protein-protein interaction (PPI) network analysis. In addition, the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were also conducted through the Database for Annotation, Visualization, and Integrated Discovery (DAVID). The molecular docking program was carried out using AutoDock 4.2 software to understand interactions between target receptors and compound ligands selected for study. We thoroughly sorted and filtered 31 pharmacologically active compounds from SNS. Subsequently, several potential target genes were predicted, of which there were 59 target genes distinctly associated with anxious insomnia. The PPI analysis indicated that the core target proteins included AKT1, IL6, TNF, SLC6A4, MAOA and GABRA2. The results of our study indicated that SNS potentially remediates anxious insomnia by reducing inflammation, neurodegeneration, and cell apoptosis of neurons. In addition, GO and KEGG enrichment analysis results indicated that SNS could modulate multiple aspects of anxious insomnia through mechanisms related to pathways of neuroactive ligand-receptor interaction. These pathways include various kinds of synaptic transmission pathways, and anti-inflammatory activity associated with response pathways. When we compared the components-compounds-targets networks and the compounds-targets-synaptic pathways networks, the five active compounds, including beta-Sitosterol, Kaempferol, Tetramethoxyluteolin, Isorhamnetin and Shinpterocarpin, were selected to conduct molecular docking experiments. Eleven target proteins, (AKT1, SLC6A4, ADRB2, MAOA, ACHE, ESR1, CYP3A4, CHRNA7, GABRA2, HTR2A and NOS3), which also play significant roles in regulating serotonergic, cholinergic, dopaminergic and GABAergic systems in the PPI network, were selected to act as receptors in molecular docking trials. The results showed that docking pairs isorhamnetin-AKT1, isorhamnetin-SLC6A4, β-sitosterol-MAOA, β- sitosterol-ACHE, isorhamnetin-CHRNA7 and shinpterocarpin-GABRA2 provided the most stable conformations of ligand-receptor binding between key compounds and core target proteins in the SNS. In the study, we offer a computational result, revealing that SNS may alleviate sleep disorders associated with anxiety through a "multi-compounds, multi-targets, and multi-pathways" mechanism. The network-pharmacology and molecular docking outcomes could theoretically confirm the anti-anxiety and anti-insomnia effects of SNS. Although this research is purely statistical and systematic without empirical validation, it serves as a stepping stone and cornerstone for subsequent experimental investigations.

Aims: The aim of the study was to investigate the pharmacological role of different chemical constituents of Paeoniae Radix Rubra (PRR) in the modulation of anticoagulation in Blood Stasis Syndrome (BSS). Background: In Traditional Chinese Medicine (TCM), BSS refers to impaired circulation or stagnation of blood flow and formation of bruises. The primary therapeutic strategy to treat BSS involves invigorating blood circulation. PRR is a widely used TCM herb for treating acute and critical diseases caused by BSS. However, the anticoagulant effects of different compounds of PRR on BSS remain elusive. Objective: This study aimed to analyze the therapeutic effect of PRR on BSS and to assess the ameliorative effect of different chemical constituents of PRR on blood circulation, clotting time, and platelet aggregation in rats with acute BSS. Methods: TCMSP (Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform) was used to screen the targets of PRR, and genes causing BSS were predicted using PharmGKB, OMIM, and TTD databases. Intersection genes between PRR and BSS targets were visualized in Venn diagrams. Core target networks of Protein-Protein Interaction (PPI) and crosstargets were constructed using Cytoscape 3.7.1, and the cross-targets were enriched using the DAVID (Database for Annotation, Visualization, and Integrated Discovery) database. Furthermore, the effects of PRR on platelet aggregation, plasma viscosity, and whole blood viscosity in the rats with BSS were examined by blood rheology and other methods. The serum levels of Endothelin-1 (ET- 1), Nitric Oxide (NO), Thromboxaneb2 (TXB2), and 6-keto-Prostaglandin F1 α (6-keto-PGF1α) in the rats were measured by the ELISA (Enzyme-Linked Immunosorbent Assay) method. Results: The main active compounds of PRR, including total glycosides, flavonoids, and polysaccharides, were identified using the TCMSP database. A total of 31 cross-targets were obtained from the intersection between 129 active targets of PRR and 345 causative genes of BSS. PPI network identified genes such as ALB (Albumin), SRC (SRC Proto-Oncogene, Non-Receptor Tyrosine Kinase), and AKT1 (AKT Serine/Threonine Kinase 1) as the core targets of PRR in alleviating BSS. Enrichment analysis showed that the common targets were mainly associated with several biological processes, including lipid and atherosclerosis, adherens junction, and focal adhesion. Following the intervention with PRR extract, the whole blood viscosity and plasma viscosity were reduced, and platelet aggregation was inhibited in the model rats in comparison to the model group. Moreover, PRR treatment also promoted thrombin time (TT), prothrombin time (PT), and Activated Partial Thromboplastin Time (APTT), increased the level of NO and 6-keto-PGF1α, but reduced the level of Fibrinogen content (FIB) and ET-1 and TXB2 in the serum of the model rats. Discussion: The present research systematically explored the anticoagulant effect of the chemical constituents of Paeoniae Radix Rubra on blood stasis syndrome in rats, applying network pharmacology analysis. Conclusion: The current findings provided a theoretical foundation for the pharmacological basis of using PRR in the management of BSS.

Jingfang granule alleviates Pseudomonas aeruginosa-induced acute lung inflammation through suppression of STAT3/IL-17/NF-κB pathway based on network pharmacology analysis and experimental validation

In this study, network pharmacology and molecular docking technology were used to explore the molecular mechanisms of the Duhuo Jisheng decoction in the treatment of osteoarthritis (OA). The chemical composition of the prescriptions was obtained from the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) database and the retrieved literature. Targets for the active ingredients were obtained using TCMSP and the Swiss Target Prediction Database. Disease targets were obtained from GeneCards and DisGeNET databases. The online tool, Venny, was used to obtain common targets for drugs and diseases. Protein-protein interactions (PPI) between common targets were analyzed using the search tool for the retrieval of interacting genes/proteins (STRING) database. Common targets were analyzed for gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment using the database for annotation, visualization and integrated discovery (DAVID) database. Molecular docking of the first 10 targets and first 10 components was verified using AutoDock Tools software, and the docking diagram was visualized using PyMOL software. After screening, 210 chemical components of the Duhuo Jisheng decoction (DHJSD) were identified. The 253 common targets of drugs and diseases were combined by eliminating repeat values. Based on PPI network analysis, the top ten targets were SRC, STAT3, MAPK3, MAPK1, RELA, PIK3R1, HSP90AA1, TP53, EP300, and AKT1. KEGG analysis showed that DHJSD could regulate the HIF-1, PI3K-Akt, and JAK-STAT signaling pathways. The biological processes involved include inflammatory reactions, the negative regulation of apoptosis, and the positive regulation of cell proliferation. Molecular docking results showed that all targets, except the RELA protein, showed good binding to the compounds, indicating that the 10 components might exert therapeutic effects by binding to the above targets. DHJSD can treat OA by regulating the HIF-1, PI3K-Akt, and JAK-STAT signaling pathways. The proteins involved were SRC, STAT3, MAPK3, MAPK1, and PIK3R1. In this study, network pharmacology was used to predict the mechanism of DHJSD in OA treatment, which was verified by molecular docking to provide experimental research ideas and scientific basis for OA treatment.

To observe the effect of vasoactive intestinal peptide (VIP) on the metabolism of intestinal fluid and cyclic AMP protein kinase A signaling pathway (cAMP-PKA) and water channel protein 3 (AQP3) in rats with constipation, and to explore the mechanism of VIP in the treatment of constipation. Methods: A total of 45 healthy adult rats were randomly divided into a control group, a model group, a model +VIP group. After 4 weeks of VIP treatment, the first black stool time were examined with the ink gastric method; the water content in feces was calculated; the morphological changes in colonic tissues were observed by HE staining. The expression of VIP and AQP3 protein levels in colon tissues were detected by Western blot; and the cAMP, PKA, AQP3 mRNA expression levels were detected by quantitative real time polymerase chain reaction (qPCR). Results: Compared with the control group, the first black stool time was prolonged, the water content of fecal decreased significantly (both P<0.01); part of the colon mucosa epithelial cells were destructed; the goblet cell volume decreased and quantity was reduced; the contents of AQP3 and VIP in colon tissues were significantly decreased, and the cAMP, PKA and AQP3 mRNA levels were decreased in the model group (all P<0.05). Compared with the model group, the first black stool time in the model +VIP group was shortened, the fecal water content increased significantly (both P<0.05); the mucosal epithelium integrity improved, the number of goblet cells increased; the content of AQP3 and VIP in colon tissues was increased, and the cAMP, PKA, and AQP3 mRNA levels were elevated (all P<0.05). Conclusion: Intravenous injection of VIP can regulate intestinal fluid metabolism and improve the symptoms of constipation in rats, which might be related to the regulation of VIP-cAMP-PKA-AQP3 signaling pathway.