Solid state fermentation process with Aspergillus kawachii enhances the cancer-suppressive potential of silkworm larva in hepatocellular carcinoma cells.

BACKGROUND: Mulberry silkworm larvae (Bombyx mori) are known as the oldest resource of food and traditional medicine. Although silkworm larvae have been reported to treat various chronic diseases, the effect of fermentation by microorganisms improving the biological activities of silkworm larvae was not reported. In the present study, fermented silkworm larvae was developed via solid-state fermentation with Aspergillus kawachii and investigated its anti-cancer activity in human hepatocellular carcinoma cells. METHODS: We investigated the anti-cancer effects of unfermented (SEE) and fermented silkworm larva ethanol extract (FSEE) on HepG2 human hepatocellular carcinoma cells as well as compared changes in free amino acid, fatty acid, and mineral contents. Anti-cancer activities were evaluated by SRB staining, cell cycle analysis, Annexin V staining, Hoechst staining, DNA fragmentation analysis and western blot analysis. Fatty acid, free amino acid and mineral contents of SEE and FSEE were determined by gas chromatography, amino acid analyzer and flame atomic absorption spectrophotometer, respectively. RESULTS: Compared with SEE, treatment with FSEE resulted in apoptotic cell death in HepG2 cells characterized by G0/G1 phase cell cycle arrest, DNA fragmentation, and formation of apoptotic bodies. Furthermore, FSEE significantly up-regulated pro-apoptotic as well as down-regulated anti-apoptotic proteins in HepG2 cells. However, an equivalent concentration of SEE did not induce cell cycle arrest or apoptosis in HepG2 cells. Moreover, fermentation process by Aspergillus kawachii resulted in enhancement of fatty acid contents in silkworm larvae, whereas amino acid and mineral contents were decreased. CONCLUSION: Collectively, this study demonstrates that silkworm larvae solid state-fermented by Aspergillus kawachii strongly potentiates caspase-dependent and -independent apoptosis pathways in human hepatocellular carcinoma cells by regulating secondary metabolites.

Anti-Pigmentation Effects of Eight Phellinus linteus-Fermented Traditional Crude Herbal Extracts on Brown Guinea Pigs of Ultraviolet B-Induced Hyperpigmentation.

We have previously found that mycelia culture broth of eight kinds of traditional herbal extracts fermented with Phellinus linteus (previously named as 8-HsPLCB) not only inhibited melanin and tyrosinase activity, but also reduced the contents of melanogenesis-related proteins, including tyrosinase and microphthalmia-associated transcription factor, in 3-isobutyl-1-methylxanthine-stimulated B16F0 melanoma cells. For a further study, the effect of 8-HsPLCB against skin pigmentation in brown guinea pigs with ultraviolet B (UVB)-induced hyperpigmentation was investigated. 8-HsPLCB (3%) and arbutin (2%) as positive controls were applied topically twice daily for 4 weeks to the hyperpigmented areas. 8-HsPLCB showed skin-lightening effect as effective as arbutin, one of the most widely used in whitening cosmetics. Melanin index values as the degree of pigmentation showed a significant reduction week by week post 8-HsPLCB treatment and then substantially reduced by 4 weeks. The degree of depigmentation after 4 weeks of topical application with 8-HsPLCB was 32.2% as compared with before treatment (0 week). Moreover, using Fontana-Masson staining and hematoxylin-eosin staining, 8-HsPLCB reduced melanin pigmentation in the basal layer of the epidermis and epidermal thickness changes exposed to the UV-B irradiation as compared with non-treatment and vehicle treatment. The intensity of the skin-lightening effect of 8-HsPLCB was similar to arbutin. These results suggest that the skin-lightening effect of 8-HsPLCB might be resulted from inhibition of melanin synthesis by tyrosinase in melanocytes. To conclude, 8-HsPLCB treatment showed reduction of the melanin pigment and histological changes induced by UV irradiation in brown guinea pigs.

Uterine arteriovenous malformation with positive serum beta-human chorionic gonadotropin: Embolization of both uterine arteries and extra-uterine feeding arteries.

The incidence of uterine arteriovenous malformation (AVM) is rare. However, it is clinically significant in that it can cause life-threatening vaginal bleeding. We report a case of a large uterine AVM with positive serum beta-human chorionic gonadotropin. A presumptive diagnosis was made; a uterine AVM accompanied by, early pregnancy or retained product of conception. Because this uterine AVM was extensive, transcatheter arterial embolization of both uterine arteries and extra-uterine feeding arteries was performed. Three months after undergoing transcatheter arterial embolization, complete resolution of the uterine AVM was confirmed without major complication.

Protective effect of cordycepin-enriched Cordyceps militaris on alcoholic hepatotoxicity in Sprague-Dawley rats.

This study was to investigate the protective effect of cordycepin-enriched Cordyceps militaris against alcohol-induced hepatotoxicity in Sprague-Dawley rats. Alcohol-feeding rats were fed diets with Paecilomyces japonica as CPJ group, C. militaris as CCM group, cordycepin-enriched C. militaris as CCMα group at the 3% (w/w) level and silymarin at the 0.1% (w/w) level for 4 weeks. Alcohol administration resulted in a significant increase in the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (γ-GTP), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) and the levels of blood alcohol and acetaldehyde in serum. However, CCMα group markedly prevented from alcohol-induced elevation of these parameters in serum. CCMα group showed the increased both hepatic activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). Unlike the action of alcohol treatment on alcoholic fatty liver, CCMα group was also attenuated lipid droplet accumulation in the hepatocytes. Present study was also confirmed the beneficial roles of silymarin (hepatoprotective agent) against alcohol-induced liver injury in rats. Therefore, cordycepin-enriched C. militaris can be a promising candidate to prevent from alcohol-induced hepatotoxicity.

Effect of fermented Angelicae gigantis Radix on carbon tetrachloride-induced hepatotoxicity and oxidative stress in rats.

This study is aimed to evaluate the protective effect of fermented Angelicae gigantis Radix (AGR) with Monascus purpureus strain on carbon tetrachloride (CCl(4))-induced hepatotoxicity and oxidative stress in rats. The activities of liver marker enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and the levels of lipid peroxidation were increased when CCl(4) was treated but these parameters were significantly decreased by fermented AGR treatment. CCl(4) treatment exhibited decrease in serum concentrations of triglyceride, total cholesterol, HDL-cholesterol, and free fatty acids, and these were also decreased by fermented AGR administration. The level of serum leptin was significantly lower in fermented AGR administration than that in normal control group. CCl(4) treatment significantly increased the concentration of liver triglyceride. The current study observed significant elevations of the thiobarbituric acid-reactive substances (TBARS) levels in the liver homogenate, mitochondrial, and microsomal fractions of CCl(4) control group compared with normal control group. CCl(4) treatment resulted in a significant decrease in the levels of plasma and hepatic glutathione, but these reductions were significantly increased by fermented AGR administration. CCl(4) induced the marked hepatocytes necrosis and fatty accumulation around the central veins. Accordingly, fermented AGR may be an ideal candidate for the hepatoprotective effect in animal model.

Antiobesity activity of fermented Angelicae gigantis by high fat diet-induced obese rats.

This study aimed to evaluate the effects of Monascus purpureus-fermented Angelicae gigantis Radix (FAG) on body weight gain, visceral fat accumulation, biochemical markers of obesity, and the mRNA expression levels of various genes involved in adipogenesis in a high-fat-diet (HFD)-induced rat model of obesity. Effect of nodakenin isolated from Angelicae gigantis on 3T3-L1 preadipocyte differentiation was also investigated in vitro. Male Sprague-Dawley rats were randomly divided into five groups (n = 6 per group) based on five dietary categories: HFD control, HFD + 2.5% (w/w) AG, HFD + 5% AG, HFD + 2.5% FAG, and HFD + 5% FAG. Present study investigated nodakenin isolated from AG and FAG roots by measuring fat accumulation in 3T3-L1 preadipocyte using Oil Red O staining. FAG administration effectively lowered the body weight gain, visceral fat accumulation, and hepatic and serum lipid and leptin concentrations in obese rats. In addition, FAG administration significantly reduced the mRNA expression levels of adipose tissue genes encoding adipocyte protein 2 (aP2), peroxisome proliferator-activated receptor γ 2 (PPARγ2), and CCAT/enhancer-binding protein α (C/EBPα) as compared with HFD group. Furthermore, nodakenin reduced the fat accumulation in differentiated 3T3-L1 adipocytes in a dose-dependent manner. FAG ameliorates HFD-induced obesity, probably by modulating multiple genes associated with adipogenesis in the visceral fat tissue of rats. Accordingly, fermented Angelicae gigantis may be an ideal candidate for obesity relief.