Alcohol, Anti-HIV Drugs, and/or Hippuric Acid Deteriorate Cellular Stresses in Senescent Hepatocytes and Aging Murine Liver
Year: 2025
Type: article
Abstract: Liver disease has increased recently in aging people living with HIV and substance use disorders, and little is known about injurious effects of anti-HIV drugs, alcohol and other substances on cellular stress responses in senescent hepatocytes and aging liver.In this study, senescence of two liver cell lines: HepG2 and AML-12, was induced by hydrogen peroxide (H 2 O 2 ) and confirmed by senescence makers including cyclin-dependent kinase inhibitor CKI (p21), senescenceassociated β-galactosidase (SA-β-gal), or insulin-like growth factor 1 (IGF1).The cell and mouse models (2-month-old versus 23-month-old) were treated with ritonavir, lopinavir, ethanol, or cocaine derivative hippuric acid.Cell stress responses and stress-related metabolic proteins were evaluated.In non-senescent cells, treatments of alcohol, ritonavir and lopinavir alone or in combinations increased expression of endoplasmic reticulum stress related glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP), had minor effects on glucose-regulated protein 75 (GRP75) and inositol 1,4,5-trisphosphate (IP3) receptor type 2 (IP3R2), ubiquitin specific protein 10 (USP10), USP17, USP20, lipogenic factor peroxisome proliferator-activated receptor gamma (PPARγ), Ras-related protein Rab-18 (RAB18), or Ras converting enzyme 1(RCE1).In the senescent cells, alcohol, ritonavir, lopinavir, and/or hippuric acid induced higher expression of GRP78, CHOP, IP3R2, USP17, USP20, PPARγ and RAB18, but reduced expression of GRP75, USP10, and RCE1.In the aged mice fed alcohol diet and the anti-HIV drugs, hepatic GRP78, CHOP, USP17, PPARγ, and triglycerides, number of senescent or dead hepatocytes, blood levels of alanine aminotransferase were significantly increased and RCE1 was reduced compared to young mice fed alcohol and the drugs.These results suggest that protein factors and responses that potentially function in ameliorating cellular stresses are undermined and protein factors that might cause cell dysfunctions or injury are exacerbated in the senescent hepatocytes and liver of aged mice treated with alcohol and anti-HIV drugs.not only worsens alcohol-induced liver injury but also exacerbates drug toxicities in aging PLWH with ART due to rising drug-drug interactions and drug modifications by alcohol [8].Besides alcohol, in PLWH with ART, certain anti-HIV drugs may also have impact on cellular senescence which may contribute to age-related decline in physiological functions.In organs other than liver, ART was reported to induce premature senescence and altered physiology in HUVECs [9,10].The anti-HIV protease inhibitors, ritonavir-boosted atazanavir, were demonstrated to induce cell growth arrest, multiple features of senescence, and senescence-associated secretory phenotypes [11].However, whether ART contribute to senescence or aging of the liver is not clear and what effects of alcohol or substances on the liver of people with ART have not been investigated.Previously, we reported that in non-senescent liver cells or non-aged animal models, the anti-HIV drug ritonavir and lopinavir induced endoplasmic reticulum (ER) and Golgi dysfunctions, cellular stress responses and hepatic injury, and combination of the HIV drugs with alcohol exacerbated the organelle stress responses and injury [12][13][14][15][16][17][18].In this study, we examined the effects of the anti-HIV drugs, combined with alcohol or a cocaine derivative, hippuric acid on cellular stress response and injury in senescent hepatocytes or aged liver of animal models, and document our findings. Materials and Methods Liver Cell Culture and TreatmentsAML12 cells (#CRL-2254) and HepG2 cells (#HB-8065) were initially purchased from ATCC (USA).AML12 cells were maintained in DMEM/Hams F-12 (#10-092-CV, Corning) supplemented with 2% of fetal bovine serum (#A3160401, Gibco, ThermoFisher), insulin-transferrin-selenium (ITS) (#41400045, Gibco), 40 ng/ml dexamethasone (#D2915, Sigma-Aldrich), and 1% of penicillinstreptomycin (#15140-122, Gibco).HepG2 cells were maintained in Dulbecco's Modified Eagle Medium (DMEM) (#11965-092, Gibco, ThermoFisher) supplemented with 10% of fetal bovine serum (FBS) (#A3160401, Gibco) and 1% of penicillin-streptomycin (#15140-122, Gibco) at 37 °C and 5% CO 2 incubator.The medium was replaced every 3 days if not otherwise stated.
Author: Cheng Ji
Institution: University of Southern California
Language: English
Subfield: Aging
Domain: Life Sciences
Open Access status: diamond
OpenAlex ID: W4409435055