Oleuropein prevents the progression of steatohepatitis to hepatic fibrosis induced by a high-fat diet in mice

Nonalcoholic steatohepatitis (NASH) is characterized by hepatocyte injury and inflammatory cell infiltration, which has been linked to peripheral insulin resistance and increased levels of triglycerides in the liver. The purposes of this study were to establish a mouse model of NASH by feeding mice a 60% high-fat diet (HFD) and to demonstrate the anti-fibrotic effects of oleuropein, which has been shown to have anti-oxidant and anti-inflammatory properties, in this HFD-induced mouse model of NASH. C57BL/6 mice were divided into three groups: a regular diet group (Chow), a HFD group and an oleuropein-supplemented HFD group (OSD), which was fed a 0.05% OSD for 6 months. The effects of oleuropein in this model were evaluated using biochemical, histological and molecular markers. The expression levels of alpha-smooth muscle actin (alpha-SMA)and collagen type I in the HFD and OSD groups were evaluated using real-time PCR and western blotting. The body weight, biochemical marker levels, nonalcoholic fatty liver disease activity score, homeostasis model of assessment-insulin resistance (HOMA-IR) and leptin levels observed in the HFD group at 9 and 12 months were higher than those observed in the Chow group. The HOMA-IR and leptin levels in the OSD group were decreased compared with the HFD group. In addition, alpha-SMA and collagen type I expression were decreased by oleuropein treatment. We established a NASH model induced by HFD and demonstrated that this model exhibits the histopathological features of NASH progressing to fibrosis. Our results suggest that oleuropein may be pharmacologically useful in preventing the progression of steatohepatitis and fibrosis and may be a promising agent for the treatment of NASH in humans.

Effect of Treadmill Exercise Training and Dietary Intake of Garcinia Cambogia Extract, Soypeptide and L-Carnitine Mixture on Body Weight Reduction in Rats Fed High-Fat Diet

This study was undertaken to examine effects of dietary intake of garcinia cambogia extract, soy peptide and L-carnitine mixture on body weight gain and obesity-related bio-markers in rats fed high-fat diet for 9 weeks with or without regular treadmill exercise. Forty 5-week-old male Sprague-Dawley rats were randomly divided into four groups; sedentary control group (SC), exercised control group (EC), sedentary formula-fed group (SF), and exercised formula-fed group (EF). The SC and EC rats were fed high-fat control diet (fat comprises 40% of total caloris), and SF and EF rats were fed high-fat formula (composed of garcinia cambogia, soy peptide and L-carnitine) supplemented diet. Statistical analyses by two-way ANOVA indicated that the regular treadmill exercise significantly lowered cumulative body weight gain, total visceral fat mass, and epididymal, perirenal and retroperitoneal fat pad weights, and serum concentrations of total cholesterol and LDL + VLDL cholesterol, insulin, c-peptide and leptin. Feeding the formula also resulted in significant reductions in cumulative body weight gain and visceral fat pad weights, along with other related parameters including serum total and LDL + VLDL cholesterol levels, and hepatic enzyme activities involved in fatty acid synthesis. Statistical analyses by one-way ANOVA revealed that the formula consumption significantly improved body weight gain (18% reduction), total visceral fat weight (20% reductions), and serum total (43% reduction) and LDL + VLDL cholesterol (54% reduction) levels, as well as serum levels of insulin (49% reduction), and c-peptide (41% reduction) in sedentary rats, but failed to exhibit significant reductions in these indices in animals under treadmill exercise program. Taken together, these results suggest that the treadmill exercise per se exhibited significant improvements in body fat reduction and other related bio-markers, and so the formula consumption did not achieve a further significant reductions in these bio-markers in exercised rats. Nevertheless, animals fed the formula with regular exercise showed the most efficient weight reduction compared to other groups either fed formula without exercise or received regular exercise without dietary supplementation.

Effect of Dietary Supplementation of Sypjeondaebotang or Jahyulyanggeuntang on Iron Bioavailability in Rats

The effect of dietary supplementation of the two oriental medicinal prescriptions, Sypjeondaebotang or Jahyulyanggeuntang, on iron bioavailability was evaluated in rats which were depleted of iron by being fed an irondeficient diet for 4 weeks. Seventy two iron-depleted rats were randomly divided into 3 groups (n = 24) , and fld one of the following experimental diets for 4 (n=8), 12 (n=8), and 25 days (n=8) : Control diet (CD), Sypjeondaebotang-supplemented diet (SD), Jahyulyanggeuntang -supplemented diet (JD). The CD contained 20 mg Fe/kg diet as FeSO4, and the SD or JD was identical except that the respective oriental medicinal prescription extract was included at the level of 4 g/kg diet. Animals fed the SD and JD for 25 days consumed significantly less food (p0.05) were lower in rats fed the SD compared to those fed the CD. Dietary supplementation of Sypjeondaebotang during the period of iron repletion significantly increased blood levels of hemoglobin (p<0.05) and hematocrit (p<0.01) measured at day 12, and increased mean corpuscular volume (p<0.05) measured at day 25, compared to the values for the CD rats. Regression analyses of hemoglobin-repletion bioassay data from rats fed the SD and JD showed the relative biological values of 123% and 99%, respectively, calculated against the slope for the CD rats. Apparent iron absorption and retention values were also significantly lower (p<0.05) in rats fed the SD for 25 days than those for the CD rats. Based on the results from diverse biochemical indices of iron status and the chemical balance study, the effect of Jahyulyanggeuntang on iron bioavailability appears to be less prominent than that for Sypjeondaebotang. Taken together, these results indicate that Sypjeondaebotang has a positive effect in restoring iron depletion by increasing the iron bioavailability in rats.

Inhibitory Effect of Cordyceps Militaris Water Extracts on Sarcoma-180 Cell-Induced Ascities Tumor in ICR Mice

This study was undertaken to evaluate the antitumor activities of Cordyceps militaris of silkworm pupa (CMP) and silkworm larva (CML), as compared with the effect of cordycepin, an active compound found in Cordyceps militaris. Antiproliferation effect of the test materials were evaluated in the sarcoma-180 cells using the MTT test. For the in vivo study, ICR mice were inoculated i.p. with 1.0 X 10(6) sarcoma-180 cells/mouse on Day 0, and were again i.p. injected with one of the following substances from Day 1 to Day 10 : saline (control group), 50 mg/kg (CMP50, CML50) ,100 ma/kg (CMP100, CML100), or 200 mg/kg (CMP200, CML200) of Cordyceps militaris water extracts, or 1 mg/kg (C1), 2 mg/kg (C2), or 4 mg/kg (C4) of cordycepin. Pretreatment of the sarcoma-180 cells with 100 mg/ml, 500 mg/ml, and 1000 mg/ml of CML (60.1+/-2.5%, 49.8+/-3.7%, and 45.4+/-0.1% of the value for untreated control cells, respectively) or CMP (68.3+/-2.1%, 55.1+/-0.9%, and 51.4+/-3.5% of the value for control cells, respectively) for 48 hrs significantly decreased the survival rate (proliferation) of tumor cells (p<0.05). Body weight of the control mice bearing ascites tumor and injected with saline was 1.4 times of the value for normal animals at day 18. Mice bearing ascites tumor and injected with cordycepin (1, 2, or 4 mg/kg) exhibited a significantly lighter body weight compared with the control mice, while animals injected with CMP or CML (50, 100, or 200 mg/kg) showed a significantly lighter body weight compared with the mice injected with cordycepin. Mice injected with CMP50, CMP100, or CMP200 mg/kg (or CML50, CML100, or CML200 mg/kg) showed a 133% (or 90%), 80% (or 62%), and 68% (or 52%) longer mean survival time, and those treated with C1, C2, or C4 exhibited a 54%, 91% and 80% longer survival time compared to the value for control mice injected with saline. These results indicate that the hot-water extracts of Cordyceps militaris of both silkworm pupa and silkworm larva have an anti-proliferation effect of tumor cells as well as the life prolongation effect in mice bearing ascites tumor, which are superior to the activities of cordycepin.