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ObjectiveTo explore the mechanism of Tangbikang granules （TBK） against diabetic peripheral neuropathy （DPN） based on network pharmacology and in-vivo experiment. MethodThe active components in medicinals of TBK and their target genes were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）. The active components of the medicinals which are not included in TCMSP were searched from previous research. After the analysis of drug-likeness by SwissADME， the target genes of them were predicted with SwissTargetPrediction. DPN-related target genes were retrieved from GeneCards. The common targets of the disease and the prescription were the hub genes of TBK against DPN， which were uploaded to Metascape for Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. High-sugar and high-fat diet and low-dose streptozotocin （STZ， ip） were employed to induce diabetes in rats， and then the model rats were respectively treated with low-dose （0.625 g·kg-1）， medium-dose （1.25 g·kg-1）， and high-dose （2.5 g·kg-1） TBK for 12 weeks. Sensory nerve conduction velocity （SNCV） was evaluated. After hematoxylin and eosin （HE） staining， the sciatic nerve was observed under light microscope to examine the nerve damage. Real-time PCR was performed to detect the gene expression of adenosine monophosphate-activated protein kinase （AMPK） pathway-related targets in rat sciatic nerve， and Western blot to measure the protein expression of AMPK and phosphorylated （p）-AMPK in rat sciatic nerve. ResultThe main active components of TBK， such as quercetin， kaempferol， β-sitosterol， leech pteridine A， stigmasterol， and baicalein were screened out， mainly acting on interleukin-6 （IL-6）， tumor necrosis factor （TNF）， protein kinase B （Akt）， JUN， and HSP90AA1 and signaling pathways such as AMPK， nuclear factor-κB （NF-κB）， and Janus kinase/signal transducer and activator of transcription （JAK/STAT）. Molecular docking results showed that β-sitosterol and stigmasterol had high binding affinity with IL-6， TNF， JUN， and HSP90AA1. As for the animal experiment， compared with the normal group， model group had low SNCV of sciatic nerve （P<0.01）， disordered and loose myelinated nerve fibers with axonotmesis and demyelinization， low mRNA expression of AMPKα， AMPKβ， peroxisome proliferator-activated receptor γ coactivator-1α （PGC-1α）， Sirtuin 3 （SirT3）， mitochondrial transcription factor A （TFAM）， and low p-AMPK/AMPK ratio in sciatic nerve （P<0.05， P<0.01）. Compared with the model group， TBK of the three doses raised the SNCV （P<0.01）， restored nerve morphology and nerve compactness， and increased the mRNA expression of AMPKα， AMPKβ， PGC-1α， SirT3， and TFAM （P<0.05， P<0.01）. The ratio of p-AMPK/AMPK in the high-dose and medium-dose TBK groups was higher than that in the model group （P<0.01）， while the protein expression in the low-dose TBK group was insignificantly different from that in the model group. ConclusionTBK exerts therapeutic effect on DPN through multiple pathways and targets. The mechanism is that it activates and regulates AMPK/PGC-1α/SirT3 signaling， which lays a basis for further study of TBK in the treatment of DPN.
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Objective: To identify potential SARS-CoV-2 3CL protease inhibitors from the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) by molecular docking approach. Methods: To alternate extensive compounds experimental screening processes, a Computer-Aided Drug Design (CADD) based molecular docking technology was performed to explore existing drug repurposing possibilities. Molecular docking model with Schrodinger suit 2018 was used to evaluate the binding abilities between TCMSP 13 143 compounds and SARS-CoV-2 3CL protease receptor-binding domain (PBD ID 6LU7), which involving in mediating viral replication and transcription functions. According to the constructed docking system, potential compounds were screened according to docking score, oral bioavailability (OB), and drug-likeness (DL). At last, a compounds-herb-target organ-function network was constructed. Results: Compared with 6LU7 original ligand docking score (-7.734), a total of 498 compounds were identified with lower docking score against 6LU7 targets. These compounds were further reduced to 60 high-priority compounds, based on OB (more than 30) and DL (more than 0.18). Meanwhile, these 60 compounds were found to interact with the amino acid residues (GLU166, GLY143, ASP187, CYS145, GLN189, LEU141, etc.) which were critically involved in the 6LU7 domain mainly by hydrogen-bonded interaction. The network exploring results revealed that these potential compounds were mainly attributed to Glycyrrhizae Radix et Rhizoma, Mori Cortex, Rhododendron dauricum, Polygoni Cuspidati Rhizoma et Radix, and Plantaginis Herba, etc., which associates with acute lung syndromes induced by SARS-CoV-2, with the effect of clearing heat and removing toxin, relieving cough and dispelling phlegm and lung-draining and relieving asthma. Conclusion: Molecular docking method provides a useful tool for the screening of SARS-CoV-2 3CL protease inhibitors from TCMSP platform.
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OBECTIVE： To study the mechanism of Wutou decoction in the treatment of osteoarthritis， and to provide a new direction and target for the treatment of osteoarthritis. METHODS： Using oral bioavailability （OB）≥30％， drug like （DL）≥0.18％ as index， active components were screened from Wutou decoction by using TCM systematic pharmacological analysis platform （TCMSP）， such as Aconitum carmichaelii， Ephedra sinica， Astragalus propinquus， Paeonia tactilora， Glycyrrhiza uralensis. Targets of osteoarthritis were obtained by retrieving therapeutic targets database （TTD） and mining thip data from gene expression database （GEO）. Target genes were analyzed by GO and KEGG pathway enrichment analysis were performed by using DAVID database. RESULTS： A total of 30 active components were screened， including quercetin， terpenoids and gardenol； 31 targets related to osteoarthritis were obtained， including β2 adrenergic receptor， arachidonate 5-lipoxygenase and androgen receptor. The biological process of Wutou decoction in treatment of osteoarthritis was mainly related to the IL-1 receptor signal transduction， synergistic activation of peroxidase proliferation activated receptor， signal transduction of tyrosine kinase receptor 2. It mainly regulated tumor necrosis factor signaling pathway， vascular endothelial growth factor signaling pathway， osteoclasts differentiation signaling pathway， nuclear factor κB signaling pathway， Toll-like receptor signaling pathway so as to play a role in the treatment of osteoarthritis. CONCLUSIONS： The study analysis the potential mechanism of Wutou decoction in the treatment of osteoarthritis based on network pharmacology， which can provide reference for further study on the material basis and target of Wutou decoction in the treatment of osteoarthritis.
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Objective To investigate the potential molecular functions and the involved signaling network of Heyan®Kuntai Capsules (HYKTc) based on the ingredient-gene targets clustering by means of bioinformatics analysis. Methods The ingredients in HYKTc were obtained by the combination of previous LC-ESI-MS/MS method and searched through Traditional Chinese Medicine System Pharmacology databases. Further the gene ontology (GO) and KEGG enrichment analysis were performed with Database for Annotation, Visualization, and Integrated Discovery (DAVID) tools. Results A total of 29 chemicals were obtained in which 21 chemicals were identified by LC-ESI-MS/MS method. Afterwards, 186 gene targets were acquired in the databases. The HYKTc-gene targets clustering were highly enriched in central nervous system, breast, and ovary. Subsequent GO analysis showed that these gene targets were significantly located in the cytosol, mitochondria and extracellular matrix, mainly functioning as lipase, kinase and oxidoreductase activity. Besides, KEGG results found that these targets were involved in the PI3K-Akt, mTOR, and insulin signaling pathways. Conclusions Using TCM databases searching combined with bioinformatics methods, the potential explanations for the clinical efficiency of HYKTc were proveded for further clinical applications.