With a Lieber-DeCarli manage liquid diet (TP 4030C), which was purchased from TROPHIC Animal Feed High-tech Co., Ltd. (Nantong, China). The energy composition of carbohydrates, protein and fat had been 19 , 18 , and 35 , respectively, from the Lieber-DeCarli ethanol liquid diet, in which ethanol supplied 28 of total calories. Nonetheless, the energy supply of ethanol was substituted by carbohydrates in the Lieber-DeCarli manage liquid diet plan. Additionally, the 95 ethanol was obtained from Guangzhou Wego Instrument Co., Ltd. (Guangzhou, China). two.3. PAK4 Inhibitor Accession experimental Design The AFLD mouse model was established depending on the approach described in a prior study [33]. Firstly, all experimental mice have been fed using the Lieber-DeCarli manage liquid diet for 5 days ad libitum. Sigma 1 Receptor Modulator Molecular Weight Subsequently, mice had been randomly divided into ethanol-fed and handle groups according to physique weight. The mixture of Lieber-DeCarli ethanol and handle liquid diets in the ratio of 1:2, 1:1, and 2:1 was treated to ethanol-fed groups, even though the manage group received the Lieber-DeCarli control liquid diet. Then, soon after 6 days, the ethanol-fed mice have been additional divided into various groups (9 mice per group), like a model group and six tea extract supplementary groups, which have been fed with the LieberDeCarli ethanol liquid diet program containing 4 (w/v) ethanol. As a consequence of the particularity of liquid feed, the mice have been divided into small cages, three in each cage. Moreover, tea extract supplementary groups were administered with diverse tea extracts like Dianhong Tea (BT1), Yingde Black Tea (BT2), Tieguanyin Tea (OT1), Fenghuang Danzong Tea (OT2), Fu Brick Tea (DT1), and Selenium-Enriched Dark Tea (DT2) at the dose of 200 mg/kg b.w. for 4 weeks, while the model and handle groups had been offered distilled water (ten mL/kg) by gavage for four weeks. Afterwards, fecal samples of every mouse were collected, and after that stored at -80 C till processing. Then, all of the mice of various groups fasting for 9 h had been weighed, anesthetized, and sacrificed to gather blood and liver samples utilised for further experiments. All experimental procedures involving animals within this study had been approved by “Principles of Care and Use of Laboratory Animals” in the College of Public Wellness, Sun Yat-Sen University (approval quantity: 2019-002; 28 February 2019).Foods 2021, ten,four of2.four. Serum TG, TC Levels, and Aminotransferase Activities Measurement After the collected blood sample kept at area temperature for a single hour, the serum sample was obtained by centrifugation at 4000g at 4 C for 10 min. Subsequently, the activities of serum aspartate transaminase (AST) and alanine transaminase (ALT), along with the levels of serum total cholesterol (TC) and triglyceride (TG) in each group had been, respectively, determined by the automated biochemistry analyzer (Roche, Mannheim, Germany). Moreover, the kits of serum ALT, AST, TG, and TC were bought from Roche diagnostics (Shanghai, China). 2.5. Hepatic Tissue Staining Just after mice have been sacrificed, the liver tissues had been removed quickly and fixed with four paraformaldehyde for two days. The liver histological evaluation was performed to evaluate liver injury including hepatocyte lipid accumulation adjust, the infiltration of inflammatory cells and degeneration using Hematoxylin-eosin (H E) staining. The liver samples were embedded in paraffin and processed to prepare 5 paraffin sections for Hematoxylin-eosin (H E) staining. Furthermore, the pictures have been captured via a light microscope (Leica,.
