ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Euscaphis konishii Hayata is a traditional medicinal plant in China, and its leaves are usually used to make dishes for hepatic or gastrointestinal issues by Chinese She nationality. Pharmacological analysis showed that E. konishii leaves contain high levels of flavonoids and chromones with favorable anti-hepatoma effect. AIM OF THE STUDY: The extract from E. konishii leaves was detected to evaluate its chemical composition, and the alcoholic liver injury mice model was adopted to elucidate its hepatoprotective effects. MATERIALS AND METHODS: The total leaf extract from E. konishii was separated by polyamide column to get the flavonoid and chromone-rich extract (FCE). Single compounds from FCE was purified by gel and Sephadex LH-20 chromatography and analyzed by nuclear magnetic resonance (NMR). The chemical component of FCE was confirmed and quantified by HPLC-MS. The OH·, O2-, DPPH and ABTS + free radical assays were adopted to estimate the antioxidant activity of FCE in vitro. The alcohol-fed model mice were established to assess the hepatoprotective capacity of FCE in vivo, through biochemical determination, histopathological analysis, mitochondrial function measurement, quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) detection and Western blot determination. RESULTS: 8 flavonoids and 2 chromones were recognized in the FCEextract by both NMR and HPLC-MS. FCE represented strong free radicals scavenging activity in vitro. With oral administration, FCE (50, 100 and 200 mg/kg) dose-dependently decreased the serum levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in alcohol-fed mice. FCE gradually reduced the malondialdehyde (MDA) content, increased the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the alcohol-treated liver tissues. FCE also alleviated the hepatic inflammation, inhibited the hepatocyte apoptosis and lessened the alcohol-induced histological alteration and lipid accumulation in the liver tissues. FCE administration inhibited the overexpression of endoplasmic reticulum (ER) chaperones signaling and unfolded protein response (UPR) pathways to defense the ER-induced apoptosis. Pretreatment with FCE also restored the mitochondrial membrane potentials andadenosine triphosphate (ATP) levels, which in turn suppressed the Cytochrome C release and mitochondria-induced apoptosis. CONCLUSIONS: FCE conferred great protection against alcoholic liver injury, which might be associated with its viability through suppressing reactive oxygen species (ROS) stress and hepatocyte apoptosis.
Subject(s)
Chemical and Drug Induced Liver Injury , Flavonoids , Alanine Transaminase , Animals , Antioxidants/pharmacology , Aspartate Aminotransferases , Chemical and Drug Induced Liver Injury/drug therapy , Chemical and Drug Induced Liver Injury/metabolism , Chemical and Drug Induced Liver Injury/prevention & control , Chromones/pharmacology , Female , Flavonoids/pharmacology , Flavonoids/therapeutic use , Liver , Mice , Oxidative Stress , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Extracts/therapeutic useABSTRACT
Liver fibrosis is a crucial pathological process involved in the hepatogenic morbidity and mortality. The pericarp of Euscaphis konishii Hayata is usually used in the cooking soup to improve liver function in Southern China, and high level of phenolic compounds has been found in the E. konishii pericarp. The total phenolic compounds extracted from E. konishii pericarp (TPEP) was obtained by polyamide column chromatograph, and 9 phenolic compounds of TPEP were identified through LC/MS and NMR. TPEP exhibited strong free radicals scavenging activity in vitro, and the chronic CCl4-induced liver fibrosis mice were established to elucidate the hepatoprotective mechanism of TPEP in vivo. TPEP treatment (50, 100 and 200 mg/kg) ameliorated the oxidative stress, immune dysfunction, inflammatory response and hepatic fibrosis induced by CCl4 introduction, alleviated the histopathological alteration and hepatocyte apoptosis in the liver tissue. Pretreatment with TPEP suppressed the activation of NADPH oxidase (NOX) signaling to attenuate oxidative stress in the liver tissue. TPEP administration inhibited the translocation of NF-κB into the nucleus to prevent the expression of downstream proinflammatory cytokines. TPEP treatment downregulated the activation of TGF-ß/Smad pathway, and facilitated the degradation of extracellular matrix through enhancing matrix metalloproteinases (MMPs) activity and decreasing the expression of matrix metalloproteinase inhibitors (TIMPs). In conclusion, TPEP inhibited CCl4-induced hepatic fibrosis through its antioxidant, anti-inflammatory and anti-fibrotic activities.
Subject(s)
Liver Cirrhosis/prevention & control , Magnoliopsida/chemistry , Phenols/pharmacology , Plant Extracts/pharmacology , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/isolation & purification , Anti-Inflammatory Agents/pharmacology , Antioxidants/administration & dosage , Antioxidants/isolation & purification , Antioxidants/pharmacology , Carbon Tetrachloride , Disease Models, Animal , Dose-Response Relationship, Drug , Free Radical Scavengers/administration & dosage , Free Radical Scavengers/isolation & purification , Free Radical Scavengers/pharmacology , Male , Matrix Metalloproteinases/metabolism , Mice , Mice, Inbred ICR , NF-kappa B/metabolism , Oxidative Stress/drug effects , Phenols/administration & dosage , Phenols/isolation & purification , Plant Extracts/administration & dosageABSTRACT
BACKGROUND: Liver injury has been recognized as a primary cause of hepatic morbidity and mortality. Euscaphis konishii Hayata, also called Euscaphis fukienensis Hsu, is usually used as a detumescent and analgesic agent to improve liver function in South China, but its mechanism of action and chemical composition are unclear. OBJECTIVE: The main aim of the study was to investigate the constituent and potential hepatoprotective mechanism of the total triterpenes of E. konishii pericarp (TTEP). METHODS: The constituent of TTEP was analyzed by a series of silica gel column to get single compounds and then identified by NMR and MS. In vitro assays were conducted to test the free radical scavenging activity of TTEP. The BCG/LPS-induced immunological livery injury mice model was established to clarify the hepatoprotective effect of TTEP in vivo. RESULTS: 8 pentacyclic triterpene acids were separated and identified by NMR and MS. TTEP treatment (50, 100, and 200 mg/Kg) improved the immune function of the BCG/LPS-infected mice, dose-dependently alleviated the BCG/LPS-induced inflammation and oxidative stress, and ameliorated the hepatocyte apoptosis in the liver tissue. CONCLUSION: The pericarp of E. konishii may be further considered as a potent natural food for liver disease treatment.