Alcoholic liver disease (ALD) is a major liver disease worldwide and can range from simple steatosis or inflammation to fibrosis/cirrhosis, possibly through leaky gut and systemic endotoxemia. Many patients with alcoholic steatohepatitis (ASH) die within 60 days after clinical diagnosis due to the lack of an approved drug, and thus, synthetic and/or dietary agents to prevent ASH and premature deaths are urgently needed. We recently reported that a pharmacologically high dose of pomegranate extract prevented binge alcohol-induced gut leakiness and hepatic inflammation by suppressing oxidative and nitrative stress. Herein, we investigate whether a dietary antioxidant ellagic acid (EA) contained in many fruits, including pomegranate and vegetables, can protect against binge alcohol-induced leaky gut, endotoxemia, and liver inflammation. Pretreatment with a physiologically-relevant dose of EA for 14 days significantly reduced the binge alcohol-induced gut barrier dysfunction, endotoxemia, and inflammatory liver injury in mice by inhibiting gut dysbiosis and the elevated oxidative stress and apoptosis marker proteins. Pretreatment with EA significantly prevented the decreased amounts of gut tight junction/adherent junction proteins and the elevated gut leakiness in alcohol-exposed mice. Taken together, our results suggest that EA could be used as a dietary supplement for alcoholic hepatitis patients.

Vanillic acid (VA), namely 4-hydroxy-3-methoxy benzoic acid, is one of the major compounds isolated from the bioactive fraction of Viticis Fructus. VA is derived from dihydroxybenzoic acid, an oxidized form of vanillin. Although studies on the antioxidant, anti-inflammatory, and anticancer pharmacologic properties of VA have been conducted, there is little research on the effect of VA on bone metabolism. The objective of the present study was to investigate the potential anti-osteoporotic properties of VA on the differentiation and mineralization in osteoblastic MC3T3-E1 cells. These cells were cultured in 0, 0.1, 1, 10 μg/mL VA for 3, 6, and 9 days. The MTT assay results showed that VA had not changed the osteoblastic cell proliferation. Extracellular alkaline phosphatase (ALP) activity increased as VA concentration increased at 9 days. ALP staining was also elevated as VA concentration increased at 6 and 9 days. Markedly, VA significantly increased the mineralized nodules in a dose-dependent manner at 3, 6, and 9 days. In addition, VA significantly increased the expression of the bone differentiation marker ALP at 3 days. The expression of the bone-specific transcription factor Runx2 protein was also elevated in the VA-treated osteoblastic MC3T3-E1 cells at 3 days. The expression of the downstream regulator of bone morphogenetic protein 2 (BMP-2) signaling, phosphorylation of ERK (p-ERK) was slightly elevated in VA-treated osteoblastic MC3T3-E1 cells at 3 days. Our results suggest that VA regulates osteoblast differentiation and mineralization in osteoblastic MC3T3-E1 cells. Taken together, our results clearly demonstrate that VA may be useful in preventing osteoporosis through the stimulation of osteoblastic differentiation and mineralization.