viously described [52]. two.5. Western Blot Analysis On day 8 of cell differentiation, whole cell protein lysates from differentiated cells were prepared, resolved by ten sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and transferred to nitrocellulose membranes. Target proteins, including phospho-protein kinase B (P-Akt), Akt, phospho-extracellular signal-regulated kinase (P-ERK), ERK, phospho-cJun N-terminal kinase (P-JNK), JNK, phospho-P38 (P-P38), P38, peroxisome proliferatoractivated receptor gamma (PPAR-), CCAAT/enhancer-binding protein alpha (C/EBP-), C/EBP-, glucocorticoid receptor (GR), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), were detected applying key antibodies and horseradish peroxidase-labeled anti-rabbit secondary antibodies (Cell Signaling Technology, Danvers, MA, USA). Target proteins had been visualized applying ECL Plus Western blotting detection reagents (GE Healthcare, Piscataway, NJ, USA). Protein levels had been determined densitometrically working with a chemiluminescence method (FUSION Solo, PEQLAB Biotechnologie GmbH, Erlangen, Germany), as previously described [53]. two.6. Statistical Analysis Statistical significance was determined utilizing one-way analysis of variance and multiple comparisons with Bonferroni correction. Statistical significance was set at p 0.05. All analyses had been performed utilizing SPSS Statistics ver. 19.0 (SPSS Inc., Chicago, IL, USA).two.6. Statistical AnalysisBiomolecules 2021, 11,Statistical significance was determined employing one-way evaluation of variance and many comparisons with Bonferroni correction. Statistical significance was set at p 0.05.21 five of All analyses were performed utilizing SPSS Statistics ver. 19.0 (SPSS Inc., Chicago, IL, USA). three. Final results three. Outcomes three.1. Network Pharmacology Evaluation three.1. Network Pharmacology Analysis three.1.1. Target Prediction and Screening of Prospective D1 Receptor Inhibitor Gene ID targets 3.1.1. Target Prediction and Screening of Prospective Targets The SwissTargetPrediction database was utilised to predict the targets of Calcium Channel Inhibitor drug hispidulin as well as the SwissTargetPrediction database was employed to predict the targets of hispidulin p-synephrine. In data preprocessing, 103 and 32 verified targets of hispidulin and and and p-synephrine. In data preprocessing, 103 and 32 verified targets of hispidulin psynephrine, respectively, were screened. In addition, 94899489 obesity-related targets were p-synephrine, respectively, have been screened. Additionally, obesity-related targets were acquired from the GeneCards database, as well as the relevance score was employed as a cut-off value. acquired in the GeneCards database, and also the relevance score was made use of as a cut-off Depending on the relevance score, 1897 obesity-related targets belonging towards the leading the top rated 20 worth. According to the relevance score, 1897 obesity-related targets belonging to 20 had been applied for thefor the analysis. As shown in Figure 1, the predicted targets of hispidulin and had been utilized evaluation. As shown in Figure 1, the predicted targets of hispidulin and psynephrine shared 53 and 23 targets, respectively, with obesity-related targets. Therefore, these p-synephrine shared 53 and 23 targets, respectively, with obesity-related targets. As a result, targets targets were selected as potential targets (Tables2).and 2). these were selected as prospective targets (Tables 1 andFigure Venn diagrams of predicted targets of compounds and obesity-related targets. (A) Venn Figure 1.1. Venndiagrams of predicted targets of compounds and obesity-related targets. (A) Venn diagram of hispidulin-predi
