| Research background: With the accelerating rate of population aging,liver cell damage-mediated lipid metabolism disorders caused by the aging process,such as diabetes,hyperlipidemia,and myocardial infarction,have become common causes of death in the elderly.Objective To explore the effect of TSPJ on the senescence of liver cells in the aging process,and explore its possible molecular mechanism.Methods(1)Animal model: rats were divided into 3M group,22 M group,TSPJ low-dose group(10mg/kg)and TSPJ high-dose group(40mg/kg).The TSPJ treatment group started feed administration at the age of 16 months and was killed after urethane anesthesia at the age of 22 months.The expressions of TG,ALT,AST were detected,and the liver lipid deposition of each group was observed by H&E staining and oil red O staining;real-time Fluorescence quantitative PCR was used to detect the expression of liver miR-33-5p,SREBP-1c,FASN,and Ch REBP m RNA,and western blot was used to detect the expression of SREBP-1c protein;Real-time fluorescent quantitative PCR and western blot to detect the expression of aging-related genes(P21,P53)and inflammatory factors(TNFα,IL-6);(2)Cell experiment: establish hepatocyte senescence model under 2% low serum status,β-Gal staining to detect the proportion of senescent cells,western blot to detect the protein expression of SREBP-1c,P21,TNFα,IL-1β,and the intervention of TSPJ;(3)Luciferase reporter gene verified the targeting relationship between miR-33-5p and SREBP-1c,western blot was used to detect the expression of SREBP-1c and total saponins of Panax japonicus after transfection of miR-33-5p mimics and miR-33-5p inhibitors Intervention.Results(1)Compared with the 3M group,the expression of TG,ALT,AST in the 22 M group were significantly increased,and the liver lipid deposition was obvious in HE staining and Oil Red O staining,and the expression of SREBP-1c,FASN,and Ch REBP increased,miR-33-5p The amount of expression decreased.The expression of aging-related genes and inflammatory factors increased,suggesting the senescence phenotype of liver cells in the aging process,as well as lipid metabolism disorders and inflammation.After treatment with TSPJ,compared with the 22 M group,the expression of TG,ALT,AST decreased significantly,the lipid deposition of HE staining and oil red O staining was significantly improved,the expression of SREBP-1c,FASN,Ch REBP genes decreased,and miR-33-5p expression increased.The decrease in the expression of aging-related genes and inflammatory factors suggests that TSPJ can improve the aging phenotype of hepatocytes in the aging process,as well as lipid metabolism disorders and inflammation.(2)In the hepatocyte senescent cell model established by low serum,the proportion of β-Gal stained senescent cells increased,and the expression of SREBP-1c,P21,TNFα,and IL-1β protein increased significantly.After TSPJ intervention,the proportion of β-Gal stained senescent cells decreased,SREBP-1c,P21,TNFα,IL-1β protein expression decreased significantly,suggesting that TSPJ can improve the senescence phenotype of liver cells in the aging process,as well as lipid metabolism disorders and inflammation.(3)The successful construction of a luciferase reporter gene plasmid containing SREBP-1c 3’-UTR confirmed that miR-33-5p can target binding to SREBP-1c;the protein expression of SREBP-1c decreased after miR-33-5p mimics was transfected;The expression of SREBP-1c increased after transfection of miR-33-5p inhibitors,which further proved that miR-33-5p and SREBP-1c have a direct target regulation relationship,and TSPJ may have an interventional effect.Conclusion There is an aging phenotype of liver cells in the aging process,accompanied by lipid metabolism disorders and inflammation;TSPJ improves liver cell aging during aging,and its mechanism may be related to the regulation of miR-33-5P/SREBP-1c signaling pathway by TSPJ and improving liver cell lipids Metabolism is related to inflammation. |
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