Metformin and Probiotics Interplay in Amelioration of Ethanol-Induced Oxidative Stress and Inflammatory Response in an In Vitro and In Vivo Model of Hepatic Injury

Figure 4

(a) MDA estimation in HepG2 cells by HPLC method. Here, 100 mM ethanol-exposed HepG2 cells were treated with 100 μl/ml probiotic V and 1 mM Met alone or together as probiotic V and Met combination for 48 h. The amount of MDA was quantified with reference to the standard area under the curve (AUC), and values represent the of three individual experiments. Statistical significance was assessed by one-way ANOVA followed by Tukey post hoc test. Statistical analysis: ^#### compared with control group; and compared with ethanol group; ^AA,BB compared with probiotic V and Met combinatorial group; (b) HPLC chromatograms of cells MDA after DNPH derivatization are shown as follows (A) MDA standard, (B) control, (C) 100 mM ethanol, (D) 100 mM ethanol+1 mM Met, (E) 100 mM ethanol+10 μl/ml probiotic V, and (F) 100 mM ethanol+10 μl/ml probiotic V and 1 mM Met combination. HepG2 cells were treated with ethanol alone or along with probiotic V and Met alone or together as probiotic V and Met combination for 48 h. After 48 h of incubation, hydrolyzed cells were mixed with DNPH and analyzed by HPLC using the ODS2 reverse-phase column. Acetonitrile and autoclaved milli-Q water containing 0.2% acetic acid in the ratio 38 : 62, respectively, was used as mobile phase.

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