Article Figures & Data
Figures
- Fig. 1.
Histologic and pathologic changes in ethanol binge treated rats. Ethanol three-repeat binge treatments (each 5 g/kg, 12 hours apart) in male and female rats (groups of four) were performed as described under Materials and Methods. Four hours after the last binge, blood samples and liver tissues were collected. (A) Blood ethanol concentration. (B) Histology image showing differences in lipid vesicles. (C) Count of lipid vesicles. (D) Triglyceride levels. (E) Serum ALT levels. (F) Cleaved caspase-3. Values are mean ± S.E.M. (n = 4 rats). For (A and C–E), the values are shown on y-axis. For (F), values are presented as fold increase over respective sex controls. FC, female control; FE, female ethanol binge; MC, male control; ME, male ethanol binge. a,Significant compared with control (P < 0.05); b, significant compared with the male ethanol binge group; (P < 0.05).
- Fig. 2.
Levels of CYP2E1 (A) and catalase (B). After binge treatments (as in Fig. 1), the whole-cell extracts were prepared (see Materials and Methods). The protein levels of these components were determined by Western blotting. Fold change over respective male or female control values are presented as mean ± S.E.M. (n = 4 rats). a, Significant compared with control (P < 0.05); b, significant compared with the male ethanol binge group; (P < 0.05). FC, female control; FE, female ethanol binge; MC, male control; ME, male ethanol binge.
- Fig. 3.
Changes in MAPK signaling components. Levels of total ERK 1/2, phospho-ERK 1/2, and ratio of phospho- to total ERK 1/2 (A); levels of total p38, phospho-p38, and ratio of phospho- to total p38 (B); and levels of Total JNK, phospho-JNK, and ratio of phospho- to total JNK (C) are presented. The experimental details were as described in Fig. 2. The protein levels of these components in whole-cell extracts were determined by Western blotting. As in Fig. 2, the values are mean ± S.E.M. (n = 4 rats). a, Significant compared with control (P < 0.05); b, significant compared with the male ethanol binge group; (P < 0.05). FC, female control; FE, female ethanol binge; MC, male control; ME, male ethanol binge.
- Fig. 4.
Changes in transcription related factors. Protein levels of total CREB (A); phospho-CREB (B); ratio of phospho- to total CREB (C); PGC-1α (D); c-FOS (E); and c-MYC (F) are shown. The protein levels of these components were determined by Western blotting. As in Fig. 2, values are fold change over respective sex controls and are mean ± S.E.M. (n = 4 rats). a, Significant compared with control (P < 0.05); b, significant compared with the male ethanol binge group; (P < 0.05). FC, female control; FE, female ethanol binge; MC, male control; ME, male ethanol binge.
- Fig. 5.
Histone H3 acetylation and phosphorylations. Levels of H3AcK9 (A); H3AcK9PS10 (B); H3PS10 (C); and H3PS28 (D) are presented. The experimental details were as described in Fig. 2. The levels of these components were determined by Western blotting. Values are fold change over respective sex control and are mean ± S.E.M. (n = 4 rats). a, Significant compared with control (P < 0.05); b, significant compared with the male ethanol binge group; (P < 0.05). FC, female control; FE, female ethanol binge; MC, male control; ME, male ethanol binge.
- Fig. 6.
Analysis of PEth species. Lipids were extracted from liver samples and PEth analyzed by ESI-MS/MS on a triple-quadrupole instrument using shotgun lipidomics method (see Materials and Methods). Values (mean ± S.D.) are from four male or female rats and are presented as picomoles per milligram of tissue.
- Fig. 7.
DGKα protein levels. In whole-cell extracts, DGKα protein levels in WCE were determined by Western blotting, followed by densitometric quantification. Increases in the protein levels after binge ethanol are presented as fold increases over respective sex controls. The experimental details were as described in Fig. 2. Values are mean ± S.E.M. (n = 4 rats). a, Significant compared with control (P < 0.05); b, significant compared with the male ethanol binge group; (P < 0.05). FC, female control; FE, female ethanol binge; MC, male control; ME, male ethanol binge.
Tables
Category Parameter Show significant differences in sexes Histology BAC, ALT, TG, lipid vesicles Catalase TOTAL CREB, phospho-CREB, c-fos, PGC1α, Phospho-ERK 1/2, phospho-p38 Diacylglycerol kinase α Show change but negligible differences in sexes Histone H3AcK9, H3AcK9PS10 CYP2E1, c-caspase 3 Total ERK1/2, total JNK Phosphatidylethanol Show no change and no differences in sexes Histone H3SP10, H3SP28 c-MYC Phospho-JNK, total p38 MAPK ALT, alanine aminotransferase; BAC, blood alcohol concentration; CREB, cyclic AMP response element binding protein; CYP2E1, cytochrome P4502E1; ERK, extracellular signal-regulated kinase; H3AcK9, histone H3 acetylated at Lys 9; H3PS10, histone H3 phosphorylated at Ser10; H3PS28, histone H3 phosphorylated at ser28; JNK, C-jun N-terminal kinase; MAPK, mitogen-activated protein kinase; PGC1α, peroxisome proliferator-activated receptor γ-coactivator 1α; TG, triglyceride.
