xation in 4 formaldehyde solution. two.3.3. Determination of High-Fat Indicators in Plasma. The frozen serum samples had been thawed at four and after that rewarmed at room temperature. The levels of TC, TG, LDL-C, HDL-C, and oxidized low-density lipoprotein (oxLDL) were determined using a microplate reader following the instructions with the kit. two.3.four. Liver Tissue Morphology Analysis. The well-fixed tissue specimens have been routinely dehydrated, embedded in paraffin, cut into 4-6 m sections, and stained employing Hematoxylin and Eosin (H E) for morphological observation with an optical microscope. The BA200 Digital trinocular microscope camera method was utilized to collect pictures. Every slice was initially observed in 40 times magnification, then, IL-5 Antagonist MedChemExpress 400-fold photos have been collected to analyze the specific liver lesions in rats. 2.4. Qualitative UPLC-QE-MS/MS Evaluation. A specific amount of freeze-dried PCE powder was weighed, dissolved in 70 methanol, ultrasonically treated for 40 minutes, allowed to cool to room temperature, after which centrifuged at 5000 rpm for five minutes. A 1.0 mL supernatant was taken and filtered with 0.22 m microporous membrane, and also the filtrate was additional diluted by methanol to a concentration of 0.2 mg/mL to receive a sample of PCE for subsequent sample injection evaluation. For qualitative analysis, a Thermo Scientific Q Exactive Orbitrap HRMS (Thermo Fisher Scientific, Massachusetts, USA) was connected to a Thermo Scientific Vanquish UPLC (Thermo Fisher Scientific, Massachusetts, USA). Chromatographic separation was accomplished on a Thermo ScientificTM AccucoreTM C18 (three one hundred mm, two.6 m) in a thermostatically controlled column compartment (30 ) [11].4 two.9. Drug-Active Ingredient-Target-Disease Network Construction. R language was utilized to construct the drug-active ingredient-target-disease data pairs, which had been imported into Cytoscape computer software to draw the drug-active ingredient-target-disease network diagram. Within the network diagram, nodes represent drug components and targets, and edges represent the correspondence involving nodes. Additionally, the network parameters were analyzed, like degree, average shortest path length, Bcl-xL Modulator review betweenness centrality, and closeness centrality in the node. Plus the significance in the node inside the network graph was also evaluated. two.10. KEGG and GO Evaluation. Functional annotation and enrichment analysis on target genes were performed employing the clusterProfiler toolkit of R language software, and the KEGG and GO functional enrichment analyses of overlapping genes had been completed. The species were set as human, as well as the enrichment result of P 0:05 was deemed as statistically substantial. Also, associated histograms and bubble charts had been supplied. two.11. In Vitro Experiments two.11.1. Cell Culture and Processing. Human hepatocellular carcinoma cell line HepG2 was purchased from Beijing Bena Biological Firm (Beijing, China) and cultured at 37 in a humidified atmosphere of five CO2 and 95 air within a sterile DMEM with 10 FBS and supplemented with 100 U/mL penicillin and one hundred U/mL streptomycin. 2.11.two. Cell Viability Test. The CCK-8 was used to detect the impact of PCE on HepG2 cells. In brief, cells have been seeded into 96-well plates (1 104 /well) and cultured at 37 for 12 hours. Then, the cells were treated with diverse doses of PCE (0, 5, 10, 20, 40, 60, 80, and 100 g/mL) and cultured in the medium at 37 for 24 and 48 hours [12], and ten L CCK-8 was added to every single properly and incubated for 1 hour. Also, HepG2 cells i