Fig 1.
General observation and morphology of gastro-intestines after acute alcohol exposure (H.E. staining).
Berberine (BBR) was administered at three different doses of 75, 150 and 300 mg/kg. A 60% alcohol was employed. The alcohol was administered at dose of 0.15 ml/10g body weight. The arrows indicate congestive necrosis places. (A): Observation of stomach and small intestines. Intestinal congestion occurs in the duodenum in model mouse. (B): Morphology of small intestines after alcohol administration. In the mouse model, small intestinal mucosa appears necrosis. BBR could prevent alcohol injury from the intestines. (C): Morphology of stomach after alcohol exposure. (D): Mucosa of small intestines (magnified 200 times). Alcohol causes gastric mucosal injury, edema with light staining. (E): Statistical score of the histopathological diagnoses for small intestines injury after alcohol consumption. Kruskal-Wallis chi-squared = 24.0696, df = 4, P = 7.735e-05. Data are expressed as the mean ± S.D. from six different mice. ###, vs. normal mice, P < 0.001. **, vs. model mice, P < 0.01.
Fig 2.
The concentration of alcohol and activity of ADH enzyme in blood of mice.
(A): Alcohol, Kruskal-Wallis chi-squared = 26.6366, df = 4, P = 2.354e-05. (B): ADH. Kruskal-Wallis chi-squared = 14.0461, df = 4, P = 0.007149. Data are expressed as the mean ± S.D. from six different mice. #, ## vs. normal mice, P < 0.05, P < 0.01. *, ** vs. model mice, P < 0.05, P < 0.01.
Fig 3.
The expressions of inflammatory cytokines of mouse stomach after acute alcohol exposure.
The concentration of alcohol was 60%. (A–G): mRNA expressions using real time PCR assay. (H–N): The expressions of protein using western blot assay. Berberine (BBR) was administered at three different doses of 75,150 and 300 mg/kg. Data are expressed as the mean ± S.D. from six different mice. ## vs. normal mice, P < 0.01. *, ** vs. model mice, P < 0.05, P < 0.01.
Fig 4.
The expressions of inflammatory cytokines of mouse small intestines after acute alcohol exposure.
The concentration of alcohol was 60%. (A–G): mRNA expressions using real time PCR assay. (H–N): The expressions of protein using western blot assay. Berberine (BBR) was administered at three different doses of 75,150 and 300 mg/kg. Data are expressed as the mean ± S.D. from six different mice. ## vs. normal mice, P < 0.01. *, ** vs. model mice, P < 0.05, P < 0.01.
Fig 5.
Expressions of inflammatory cytokines after berberine (BBR) on Caco2 cells injured by alcohol exposure in vitro.
(A–E): mRNA expressions using real time PCR assay. (F–J): The expressions of protein using western blot assay. Alcohol was used at concentration of 348 mmol/L. BBR was administered at concentration of 5.95 μmol/L. (K): Cellular viability after alcohol exposure. (L): Cellular viability after BBR administration. Data are expressed as the mean ± S.D. from six independent experiments. #, ## vs. the control, P < 0.05, P < 0.01. ** vs. the model, P < 0.01.
Fig 6.
Effect of berberine (BBR) on the transcriptions of TLR2 and TLR4 on 293T cells.
(A): Constructions of TLR2, TLR4 and NOD2 plasmids. (B–D): mRNA expression of GFP. (E–G): Protein expression of GFP promoted by TLR2, TLR4 and NOD2. (H): Lanes of protein expression detected using western blot assay. (I): Cellular viability after alcohol exposure. (J): Cellular viability after BBR administration. NS: no significance. Alcohol was used at concentration of 44 mmol/L. BBR was administered at concentration of 1.49 μmol/L. Data are expressed as the mean ± S.D. from three independent experiments. #, ## vs. the control, P < 0.05, P < 0.01. ** vs. the model, P < 0.01.