Chi Chang Huang

Protective effect of Acacia confusa bark extract and its active compound gallic acid against carbon tetrachloride-induced chronic liver injury in rats

Acacia confusa Merr. (Leguminosae), a species native to Taiwan, is widely distributed on the hills and lowlands of Taiwan, and has been traditionally used as a medicine. The hepatoprotective effects of A. confusa bark extract (ACBE) and its active constituent gallic acid were evaluated against carbon tetrachloride (CCl(4))-induced hepatotoxicity in rats. CCl(4)-induced hepatic pathological damage and significantly increased the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and malondialdehyde (MDA) in plasma, and cytochrome P4502E1 (CYP2E1) protein expression in hepatic samples, and decreased the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) in erythrocytes. Treatment with ACBE, gallic acid or silymarin could decrease significantly the AST, ALT, and MDA levels in plasma, and CYP2E1 expression in liver tissues, and increase the activities of SOD and GPX in erythrocyte when compared with CCl(4)-treated group. Liver histopathology also showed that ACBE, gallic acid or silymarin could significantly reduce the incidence of liver lesions induced by CCl(4). These results suggested that the ACBE and gallic acid exhibit potent hepatoprotection against CCl(4)-induced liver damages in rats, and the hepatoprotective effects of ACBE and gallic acid may be due to the modulation of antioxidant enzymes activities and inhibition of lipid peroxidation and CYP2E1 activation.

A galactolipid possesses novel cancer chemopreventive effects by suppressing inflammatory mediators and mouse B16 melanoma.

Crassocephalum rabens (Asteraceae) is a popular anti-inflammatory folk medicine and food supplement. We investigated the cancer chemopreventive bioactivity of C. rabens phytocompounds in vitro and in vivo using cell- and gene-based bioassays and a mouse B16 melanoma model. The bioactive glyceroglycolipid 1,2-di-O-alpha-linolenoyl-3-O-beta-galactopyranosyl-sn-glycerol (dLGG) that was identified from C. rabens was found in vitro and in vivo to be a potent nitric oxide (NO) scavenger. dLGG treatment inhibited both mRNA and protein expression of inducible NO synthase and cyclooxygenase-2 (COX-2) in murine macrophages and inhibited COX-2 gene transcription in 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated B16 cells. In immunohistochemical studies, dLGG inhibited TPA-induced expression of COX-2 and nitration of proteins in mouse skin. dLGG could also significantly inhibit lipopolysaccharide-induced prostaglandin E(2) production in murine macrophages. Furthermore, dLGG prevented nuclear translocation of cytoplasmic nuclear factor-kappaB (NF-kappaB) by suppressing IkappaBalpha phosphorylation and degradation. Structure-activity relationship study by electrophoretic mobility shift assay indicated that the dilinolenoylglycerol moiety in dLGG is the essential structural feature preventing NF-kappaB.DNA complex formation. A dLGG-enriched extract from C. rabens (10 mg/kg) markedly suppressed B16 melanoma growth in C57BL/6J mice following i.p. administration, an effect comparable with that of cisplatin, a cancer chemotherapeutic drug. This study shows the detailed molecular mechanism(s) underlying the anti-inflammatory and tumor-suppressive effects of a natural galactolipid.

Hepatoprotective phytocompounds from Cryptomeria japonica are potent modulators of inflammatory mediators.

Cryptomeria japonica is an important plantation conifer tree in Asia. This study aimed to characterize the anti-inflammatory and hepatoprotective activities of the phytocompounds from C. japonica wood on LPS- or TPA-induced activation of proinflammatory mediators and CCl(4)-induced acute liver injury in mice. A CJH7-2 fraction was purified from C. japonica extracts following bioactivity-guided fractionation, and it exhibited significant activities on inhibition of NO production and iNOS expression as well as up-regulating HO-1 expression in LPS-stimulated macrophages. CJH7-2 also potently inhibits COX-2 enzymatic activity (IC(50)=5 microg/mL) and TPA-induced COX-2 protein expression in mouse skin (1mg/200 microL/site). CJH7-2 (10 mg/kg BW) can prevent CCl(4)-induced liver injury and aminotransferases activities in mice. Chemical fingerprinting analysis showed that terpenes are the major bioactive compounds in the CJH7-2 fraction. This is the first study to demonstrate that chemical constituents from the wood extract of C. japonica possess anti-inflammatory activities in vitro and in vivo that may play a role in hepatoprotection.

Effect of intestinal microbiota on exercise performance in mice.

Abstract Hsu, YJ, Chiu, CC, Li, YP, Huang, WC, Huang, YT, Huang, CC, and Chuang, HL. Effect of intestinal microbiota on exercise performance in mice. J Strength Cond Res 29(2): 552–558, 2015—The antioxidant enzyme system helps protect against intense exercise-induced oxidative damage and is related to the physical status of athletes. Evidence suggests that intestinal microbiota may be an important environmental factor associated with host metabolism, physiology, and antioxidant endogenous defense. However, evidence of the effect of gut microbiota status on exercise performance and physical fatigue is limited. We investigated the association of intestinal bacteria and exercise performance in specific pathogen-free (SPF), germ-free (GF), and Bacteroides fragilis (BF) gnotobiotic mice. Endurance swimming time was longer for SPF and BF than GF mice, and the weight of liver, muscle, brown adipose, and epididymal fat pads was higher for SPF and BF than GF mice. The serum levels of glutathione peroxidase (GPx) and catalase were greater in SPF than GF mice. Serum superoxide dismutase activity was lower in BF than SPF and GF mice. In addition, hepatic GPx level was higher in SPF than GF and BF mice. Gut microbial status could be crucial for exercise performance and its potential action linked with the antioxidant enzyme system in athletes.

Cytoprotective Effect of American Ginseng in a Rat Ethanol Gastric Ulcer Model

Panax quinquefolium L. (American Ginseng, AG) is one of the most popular herbal medicines in the World. We aimed to investigate whether chronic (28-day) supplementation with AG could protect against ethanol-induced ulcer in gastric tissue. Furthermore, we investigated the possible molecular mechanisms leading to AG-mediated gastric mucosal protection. We randomized 32 male Wistar rats into four groups for treatment (n = 8 per group): supplementation with water (vehicle) and low-dose (AG-1X), medium-dose (AG-2X) and high-dose (AG-5X) AG at 0, 250, 500, and 1250 mg/kg, respectively. In the first experiment, animals were fed vehicle or AG treatments for 4 weeks. At day 29, 75% ethanol was given orally to each animal at 10 mL/kg to induce gastric ulceration for 2 h. In a second experiment, animals were pretreated orally with each treatment for 1 hr before a single oral administration of ethanol (70%, 10 mL/kg). Trend analysis revealed that AG treatments inhibited ethanol-induced gastric mucosal damage. AG supplementation dose-dependently decreased the pro-inflammatory levels of interleukin 1β and cyclooxygenase 2 and the expression of pro-apoptotic proteins tBid, cytochrome C, and caspases-9 and -3 and increased the levels of anti-apoptotic proteins Bcl-2, Bcl-xL and p-Bad. AG could have pharmacological potential for treating gastric ulcer.

Bioactive phytochemicals of leaf essential oils of Cinnamomum osmophloeum prevent lipopolysaccharide/ d -galactosamine (LPS/ d -GalN)-induced acute hepatitis in mice

The purpose of this study was to investigate the bioactive phytochemicals of leaf essential oils of Cinnamomum osmophloeum on lipopolysaccharide/D-galactosamine (LPS/D-GalN)-induced acute hepatitis. The results revealed that post-treatment with 100 μmol/kg trans-cinnamaldehyde, (-)-aromadendrene, T-cadinol, or α-cadinol significantly decreased the aspartate aminotransferase (AST), alanine aminotransferase (ALT), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6) levels in serum. Moreover, both T-cadinol and α-cadinol treatments decreased the expressions of cleaved caspase-3 and cleaved poly-ADP ribose polymerase (PARP) in the liver tissues when compared with the LPS/D-GalN group. Liver histopathology also showed that silymarin, trans-cinnamaldehyde, (-)-aromadendrene, T-cadinol, or α-cadinol significantly reduced the incidence of liver lesions induced by LPS/D-GalN. These results suggest that the above phytochemicals exhibit potent hepatoprotection against LPS/D-GalN-induced liver damage in mice, and their hepatoprotective effects may be due to the modulation of anti-inflammatory activities.

Metabolomics characterization of energy metabolism reveals glycogen accumulation in gut-microbiota-lacking mice

Microbiota in the gut are considered an important environmental factor associated with host metabolism and physiology. Although gut microbiota are known to contribute to hepatic lipogenesis and fat storage, little is known about how the condition influences the deposition of glycogen in the liver. To better understand and characterize the host energy metabolism in guts lacking microbiota, we compared the liver metabolome of specific pathogen-free and germ-free mice by gas chromatography-mass spectrometry combined with partial least-squares discriminant analysis. We identified 30 of 52 highly reproducible peaks in chromatograms of liver tissue extracts from the two groups of mice. The two groups showed significant differences in metabolic profile. Changes in liver metabolism involved metabolites such as amino acids, fatty acids, organic acids and carbohydrates. The metabolic profile of germ-free mice suggests that they synthesize glycogen and accumulate it in the liver through gluconeogenesis and glycogenesis. Our findings shed light on a new perspective of the role of gut microbiota in energy metabolism and will be useful to help study probiotics, obesity and metabolic diseases.

Ganoderma tsugae hepatoprotection against exhaustive exercise-induced liver injury in rats

Several studies have been shown that accelerated apoptosis is involved in post-exercise lymphocytopenia and tissue damage after high-intensity exercise. Ganoderma tsugae (GT) is one of the well-known medicinal mushrooms that possess various pharmacological functions. This mushroom has traditionally been used for health promotion purposes. This study investigates the hepatoprotective effects of GT on exhaustive exercise-induced liver damage. Twenty-four male Sprague-Dawley rats were randomly divided into four groups and designated as exhaustive exercise only (E), exhaustive exercise with low dosage (EL), medium dosage (EM) and high dosage (EH) GT at 0, 0.1875, 0.9375 and 1.875 g/kg/day, respectively. After 30 days all rats were euthanized immediately after an exhaustive running challenge on a motorized treadmill. The rat livers were immediately harvested. Evidence of apoptotic liver cell death was revealed using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and caspases mediated cascade events. DNA fragmentation, an apoptosis process, can be examined using TUNEL assay. A few TUNEL-positive hepatocytes, compared to the exercise only group, were observed in the livers from exhaustive animals supplemented with GT. Immunoblot analysis also showed that caspase-6-mediated specific cleavage of lamin A/C was increased significantly in the livers of group E, but was significantly decreased in the EM and EH groups. Our observations demonstrate that GT possesses anti-apoptotic and hepatoprotective potential after exhaustive exercise.

Effect of Exercise Training on Skeletal Muscle SIRT1 and PGC-1α Expression Levels in Rats of Different Age.

The protein deacetylase sirtuin 1 (SIRT1) and activate peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) pathway drives the muscular fiber-type switching, and can directly regulate the biophysiological functions of skeletal muscle. To investigate whether 12-week swimming exercise training modulates the SIRT1/PGC-1α pathway associated proteins expression in rats of different age. Male 3-month-old (3M), 12-month-old (12M) and 18-month-old (18M) Sprague-Dawley rats were used and assigned to sedentary control (C) or 12-week swimming exercise training (E) and divided into six groups: 3MC (n = 8), 12MC (n = 6), 18MC (n = 8), 3ME (n = 8), 12ME (n = 5) and 18ME (n = 6). Body weight, muscle weight, epididymal fat mass and muscle morphology were performed at the end of the experiment. The protein levels of SIRT1, PGC-1α, AMPK and FOXO3a in the gastrocnemius and soleus muscles were examined. The SIRT1, PGC-1α and AMPK levels in the gastrocnemius and soleus muscles were up-regulated in the three exercise training groups than three control groups. The FOXO3a level in the 12ME group significantly increased in the gastrocnemius muscles than 12MC group, but significantly decreased in the soleus muscles. In 3-, 12- and 18-month-old rats with and without exercise, there was a significant main effect of exercise on PGC-1α, AMPK and FOXO3a in the gastrocnemius muscles, and SIRT1, PGC-1α and AMPK in the soleus muscles. Our result suggests that swimming training can regulate the SIRT1/PGC-1α, AMPK and FOXO3a proteins expression of the soleus muscles in aged rats.

Hypolipidemic effects and safety of Lactobacillus reuteri 263 in a hamster model of hyperlipidemia.

We aimed to verify the beneficial effects of probiotic strain Lactobacillus reuteri 263 (Lr263) on hypolipidemic action in hamsters with hyperlipidemia induced by a 0.2% cholesterol and 10% lard diet (i.e., high-cholesterol diet (HCD)). Male Golden Syrian hamsters were randomly divided into two groups: normal (n = 8), standard diet (control), and experimental (n = 32), a HCD. After a two-week induction followed by a six-week supplementation with Lr263, the 32 hyperlipidemic hamsters were divided into four groups (n = 8 per group) to receive vehicle or Lr263 by oral gavage at 2.1, 4.2, or 10.5 × 109 cells/kg/day for 6 weeks, designated the HCD, 1X, 2X and 5X groups, respectively. The efficacy and safety of Lr263 supplementation were evaluated by lipid profiles of serum, liver and feces and by clinical biochemistry and histopathology. HCD significantly increased serum levels of total cholesterol (TC), triacylglycerol (TG) cholesterol, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), LDL-C/HDL-C ratio, hepatic and fetal TC and TG levels, and degree of fatty liver as compared with controls. Lr263 supplementation dose dependently increased serum HDL-C level and decreased serum TC, TG, LDL-C levels, LDL-C/HDL-C ratio, hepatic TC and TG levels, and fecal TG level. In addition, Lr263 supplementation had few subchronic toxic effects. Lr263 could be a potential agent with a hypolipidemic pharmacological effect.