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CTRP3 Prevents ETOH‐ Induced Hepatocyte Apoptosis
Author(s) -
Dunlay Samantha,
Peterson Jonathan M
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.734.1
Subject(s) - apoptosis , programmed cell death , hepatocyte , western blot , fatty liver , alcoholic liver disease , biology , microbiology and biotechnology , chemistry , biochemistry , medicine , disease , in vitro , gene , cirrhosis
Alcohol abuse is the third leading cause of preventable death in the United States. Our lab works with the novel protein C1q TNF Related Protein (CTRP3), which is secreted from adipose tissue and circulates in the blood stream. Our past lab experiments show that CTRP3 prevents nonalcoholic fatty alcoholic liver disease. Nevertheless, the effects of CTRP3 on alcoholic fatty liver disease (AFLD) are still unknown. Our lab to date, has determined that CTRP3 prevents alcohol‐induce hepatocyte (liver cell) death. The purpose for this project is to understand how. Previous research has demonstrated that ethanol consumption increases apoptosis (programmed cell death) in hepatocytes through an inappropriate elevation in apoptotic signaling. Our working hypothesis is that CTRP3 prevents ethanol‐induced hepatocyte cells death by suppressing ethanol‐induced elevations in pro‐apoptotic signaling. To test our hypothesis we examine the effects of ethanol plus/minus CTRP3 treatment on the Bcl‐2 family of apoptotic regulatory proteins. The Bcl‐2 family can either induce (pro‐apoptotic) cell death or inhibit cell death (anti‐apoptotic). Methods Hepatocytes were treated with ethanol (100 mM) plus/minus recombinant CTRP3 and Bcl‐2 family protein content was determined through western blot analysis. We measured the specific apoptotic regulator proteins: Bcl‐2‐associated X protein (BAX), which is pro‐apoptotic, and B‐cell lymphoma 2 (BCL‐2) and (BCL‐XL), which are anti‐apoptotic. Western blots were performed using standard techniques; briefly, protein samples are separated by size through SDS‐PAGE (PolyAcrylamide Gel Electrophoresis). The proteins are then transferred to a nitrocellulose membrane and relative protein concentrations are detected by chemiluminescence. Results ethanol treatment increases the amount of BAX and decreases the amount of BCL‐XL but not BCL‐2 protein content in the hepatocytes. However, treatment with CTRP3 stopped the ethanol‐induced elevations to BAX and reductions in BCL‐XL. In conclusion this data supports our hypothesis that CTRP3 prevents ethanol‐induced hepatic cell death. Support or Funding Information East Tennessee State University RDC: E82262 NIAAA: R03AA023612