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Objective@#To explore the molecular mechanism of resveratrol (RES) in the treatment of oral squamous cell carcinoma (OSCC) through the use of biological information methods such as network pharmacology and molecular docking and to provide a theoretical reference for the clinical application of RES in the treatment of OSCC.@*Methods@#The Swiss Target Prediction(http://www.swisstargetprediction.ch), SEA (http://sea.bkslab.org)database, and Pharm mapper database(http://lilab-ecust.cn) were used to retrieve RES-related targets, and the DISGENET (www.disgenet.org), OMIM (https://omim.org) and GeneCards (https://www.genecards.org) databases were used to screen OSCC disease targets. The intersection of drugs and disease targets was determined, and Cytoscape 3.7.2 software was used to construct a "drug-diseasetarget pathway" network. The Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database was used to construct a target protein interaction network, and the DAVID database was used for enrichment analysis of key proteins. Finally, molecular docking validation of key proteins was performed using AutoDock and PyMOL. The enrichment analysis and molecular docking results were integrated to predict the possible molecular mechanisms of RES treatment in OSCC; western blot was used to determine the effect of resveratrol at different concentrations (50, 100) μmol/L on the expression of Src tyrosine kinase (SRC), epidermal growth factor receptor (EGFR), estrogen receptor gene 1 (ESR1), and phosphatidylinositol 3 kinase/protein kinase B (PI3K/AKT) signaling pathway proteins in OSCC HSC-3 cells.@*Results@#A total of 243 targets of RES drugs and 6 094 targets of OSCC were identified. A total of 116 potential common targets were obtained by intersecting drugs with disease targets. These potential targets mainly participate in biological processes such as in vivo protein self-phosphorylation, peptide tyrosine phosphorylation, transmembrane receptor protein tyrosine kinase signaling pathway, and positive regulation of RNA polymerase Ⅱ promoter transcription, and they interfere with the PI3K/AKT signaling pathway to exert anti-OSCC effects. The docking results of resveratrol with OSCC molecules indicated that key targets, such as EGFR, ESR1, and SRC, have good binding activity. The results of cell-based experiments showed that resveratrol inhibited the protein expression of SRC, EGFR, ESR1, p-PI3K, and p-AKT in HSC-3 cells in a dose-dependent manner.@*Conclusion@#RES can inhibit the expression of its targets EGFR, ESR1, SRC, p-PI3K, and p-AKT in OSCC cells.
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OBJECTIVE@#Duranta repens is reported to contain a wide array of secondary metabolites, including α-amylase and α-glucosidase inhibitors, and - has potent antioxidant activity. The present study evaluated the network pharmacology of D. repens (whole plant) with targets related to diabetes mellitus and assessed its outcome by evaluating the effects of the hydroalcoholic extract of D. repens in streptozotocin-nicotinamide-induced diabetes mellitus in rats.@*METHODS@#Phytoconstituents of D. repens were retrieved from an open-source database and published literature, and their targets were predicted for diabetes mellitus using BindingDB and the therapeutic target database. Protein-protein interaction was predicted using STRING, and pathways involved in diabetes mellitus were identified using the Kyoto Encyclopedia of Genes and Genomes pathway browser. Druglikeness, ADMET profile (absorption, distribution, metabolism, excretion and toxicity) and cytotoxicity of compounds modulating proteins involved in diabetes were predicted using MolSoft, admetSAR2.0 and CLC-Pred, respectively. The interaction network among phytoconstituents, proteins and pathways was constructed using Cytoscape, and the docking study was performed using AutoDock4.0. The hydroalcoholic extract of D. repens was evaluated using streptozotocin-nicotinamide-induced diabetes mellitus animal model for 28 d, followed by an oral glucose tolerance test. At the end of the study, biochemical parameters like glycogen content, hepatic enzymes, antioxidant biomarkers and lipid profiles were quantified. Further, the liver and pancreas were collected for a histopathology study.@*RESULTS@#Thirty-six different secondary metabolites from D. repens were identified to regulate thirty-one targets involved in diabetes mellitus, in which protein-tyrosine phosphatase 1B (PTP1B) was primarily targeted. Enrichment analysis of modulated proteins identified 12 different pathways in diabetic pathogenesis in which the phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathway was chiefly regulated. The docking study found that durantanin I possessed the highest binding affinity (-8.9 kcal/mol) with PTP1B. Similarly, ADMET profiling showed that the majority of bioactive constituents from D. repens had higher human intestinal absorptivity and minimal cytotoxicity to normal cell lines, than tumor cell lines. Further, an in vivo animal study reflected the efficacy of the hydroalcoholic extract of D. repens to lower the elevated blood glucose level by stimulating insulin secretion, maintaining pancreatic β cell mass, regulating glycolysis/gluconeogenesis and enhancing the glucose uptake in skeletal muscles.@*CONCLUSION@#The present study reflected the probable network interaction of bioactive constituents from D. repens, their targets and modulated pathways, which identified the prime regulation of the PI3K-Akt signaling pathway and PTP1B protein. Modulation of PTP1B protein and PI3K-Akt signaling pathway could contribute to enhancing glucose uptake, insulin production and glycolysis and decreasing gluconeogenesis in diabetes, which was evaluated via the experimental study.
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Objective To observe the variation of phosphatidylinositol-3 kinase (PI3K),protein kinase B (AKT),sodium iodide symporter (NIS) mRNA and protein expression in rat mammary tissues and serum insulin growth factor Ⅰ (IGF-1) under different iodine nutrition levels,and to study the role of PI3K-AKT signaling pathway in the process of mammary gland intaking iodine during lactation period.Methods Totally 130 Wistar rats (100 female rats,30 male rats) were randomly divided into five groups with 20 female rats in each group:①control group (NI):was feed with normal diet and iodine content 50 μg/L in deionized water;②low iodine group 1 (LI1 group):was feed with low iodine diet and deionized water;③low iodine group 2 (LI2):was feed with low iodine diet and iodine content 5 μg/L in deionized water;④high iodine group 1 (HI1 group):was feed with normal diet and iodine content 3 000 μg/L in deionized water;⑤high iodine group 2 (HI2):was feed with normal diet and iodine content 10 000 μg/L in deionized water.After feeding for 3 months,females were mated with male rats,then male rats were taken out and every female rat was feed individually.Urinary iodine level of rats in lactation period 10 days after giving birth was tested.Blood and mammary tissue samples of rats in lactation period were taken after killing them.Enzyme linked immunosorbent assay (ELISA) was used to detect serum IGF-1 level,real-time fluorescence quantification PCR to detect the mRNA expression of mammary gland PI3K,AKT and NIS,Western blotting to detect mammary gland PI3K,total AKT,phosphorylation AKT (p-AKT) and NIS protein expression.Results The medians urinary iodine of lactation period rats in LI1 and LI2 (3.16,6.36 μg/L) were significantly lower than that in NI group (162.59 μg/L),and were significantly higher in HI1 and HI2 (2 356.27,11 507.29 μg/L) than that in NI group.The differences were statistically significant (all P ＜ 0.01).Compared with control group [(8.84 ± 2.12) μg/L],the content of serum IGF-1 increased significantly in lactation period rats in LI1 and LI2 groups [(13.30 ± 2.37) and (10.90 ± 1.92) μg/L,all P＜ 0.01].The real-time fluorescence quantification PCR detection results indicated that the differences were statistically significant by comparing NIS,AKT,PI3K mRNA expression of the mammary tissues of lactation period rats in the five groups (F=14.916,36.477,14.994,all P＜ 0.01).Among them,NIS mRNA expression quantities in LI1 and LI2 groups (0.75 ± 0.40,0.89 ± 0.51) were significantly higher than that in NI group (0.53 ± 0.31),and significantly lower in HI2 group (0.30 ± 0.24) than that in NI group (P ＜ 0.05 or ＜ 0.01).AKT mRNA expression quantities in LI1 and LI2 groups (0.90 ± 0.19,0.64 ± 0.22) were significantly higher than that in NI group (0.43 ± 0.22),and significantly lower in HI2 group (0.29 ± 0.15) than that in NI group (P ＜ 0.05 or ＜ 0.01).PI3K mRNA expression quantity in LI1 group (0.85 ± 0.42) was significantly higher than that in NI group (0.50 ± 0.24),and significantly lower in HI2 group (0.28 ± 0.10) than that in NI group (all P ＜ 0.01).Western blot detection results indicated that the differences were statistically significant by comparing mammary gland NIS protein expression of lactation period rats in the five groups (F=4.510,P＜ 0.01).Among them,LI1 group (1.67 ± 0.97) was significantly higher than NI group (0.87 ± 0.43,P ＜ 0.05).The differences were statistically significant by comparing the p-AKT protein expression among groups (F =3.528,P ＜ 0.05).Among them,HI2 group (1.10 ± 0.30) was significantly higher than NI group (0.75 ± 0.23,P ＜0.05).The differences were not statistically significant by comparing total AKT and PI3K protein expression among groups (F =0.558,1.319,all P ＞ 0.05).Conclusion The inhibitory effect of PI3K-AKT signaling pathways on NIS in the mammary gland was weaker than the effect of iodine intake.But the expression of functional p-AKT was gradually increased with the increment of iodine intake,which had been presented inhibit effect on NIS expression in lactating mammary gland.