ABSTRACT
This study was designed to investigate the antioxidative, antiinflammatory and metabolism-regulating effects of gastrodin (GSTD) in the treatment of nonalcoholic fatty liver disease (NAFLD). Oleic acid (OA) was used to induce steatosis in HL-7702 cells; a high-fat or high-fat and high-cholesterol diet was used to induce NAFLD in mice and rats. Our results showed that GSTD significantly increased hepatic superoxide dismutase (SOD) but decreased reactive oxygen species (ROS)/malondialdehyde (MDA) and reduced the mRNA levels of proinflammatory cytokines both in vitro and in vivo. GSTD promoted the phosphorylation of nuclear factor erythroid-2-related factor-2 (Nrf2) at serine (Ser) 40, stimulated its nuclear translocation and increased hepatic expression of heme oxygenase-1 (HO-1). GSTD activated AMP-activated protein kinase (AMPK), suppressed hepatic steatosis, lowered serum triglyceride (TG)/glucose and decreased body weight gain in animals with NAFLD. The stimulating effects of GSTD on the Nrf2 pathway as well as its antioxidative/antiinflammatory activities were abolished by compound C in OA-treated HL-7702 cells. In summary, our results demonstrate that GSTD activates the AMPK/Nrf2 pathway, ameliorates oxidative stress/proinflammatory response and improves lipid metabolism in NAFLD. Our findings may support the future clinical application of GSTD for the treatment of NAFLD to reduce hepatic steatosis, oxidative stress and proinflammatory response.
Subject(s)
AMP-Activated Protein Kinases/metabolism , Benzyl Alcohols/therapeutic use , Gastrodia/chemistry , Glucosides/therapeutic use , Inflammation/prevention & control , NF-E2-Related Factor 2/metabolism , Non-alcoholic Fatty Liver Disease/drug therapy , Oxidative Stress/drug effects , Animals , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Antioxidants/pharmacology , Antioxidants/therapeutic use , Benzyl Alcohols/pharmacology , Glucosides/pharmacology , Heme Oxygenase-1/metabolism , Hep G2 Cells , Humans , Inflammation/metabolism , Male , Malondialdehyde/blood , Mice, Inbred C57BL , Non-alcoholic Fatty Liver Disease/chemically induced , Non-alcoholic Fatty Liver Disease/metabolism , Non-alcoholic Fatty Liver Disease/pathology , Phytotherapy , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolism , Triglycerides/bloodABSTRACT
It is known that beta-adrenoceptor (AR) in the basolateral nucleus of amygdala (BLA) plays an essential role in fear memory formation. However, the cellular and subcellular distributions of beta1- and beta2-ARs in the BLA and their roles in fear memory formation are poorly understood. Here, we report that both beta1- and beta2-ARs are predominantly expressed in BLA neurons but not in astrocytes. beta1-AR is distributed in the cell membrane and cytoplasm of neurons, whereas beta2-AR is localized not only in the cell membrane and cytoplasm but also in the nucleus. Intra-BLA infusion of the beta1-AR antagonist metoprolol and atenolol or the beta2-AR antagonist ICI118551 and butoxamine produces a severe deficit in 24-h auditory fear memory, leaving 1-h memory intact. Western-blot analysis reveals that the protein level of cytoplasmic beta1-AR significantly increases 2- and 4-h postconditioning, whereas that of cytoplasmic or nuclear beta2-AR is unchanged. The present results indicate that beta1- and beta2-ARs in the BLA have differential subcellular localizations and both are required for the consolidation of auditory fear memory.
Subject(s)
Amygdala/metabolism , Fear/physiology , Memory/physiology , Receptors, Adrenergic, beta-1/metabolism , Receptors, Adrenergic, beta-2/metabolism , Acoustic Stimulation , Adrenergic beta-Antagonists/pharmacology , Amygdala/drug effects , Animals , Astrocytes/metabolism , Blotting, Western , Fluorescent Antibody Technique , Male , Microscopy, Confocal , Neurons/metabolism , Protein Transport/drug effects , Protein Transport/physiology , Rats , Rats, Sprague-Dawley , Receptors, Adrenergic, beta-1/drug effects , Receptors, Adrenergic, beta-2/drug effectsABSTRACT
OBJECTIVE: To analyze the chemical constituents of supercritical carbon dioxide extraction products from Cnidium monieri. METHOD: Four-factor and three-level orthogonal experimental design was used to optimize the SFE conditions as guided by the content of total coumarins in the extract. The chemical constituents were separated and identified by recrystalization. RESULT: Optimum extraction process was established: 25 MPa as extraction pressure, 50 degrees C as extraction temperature, 6.5 MPa as separation pressure and 60 degrees C as separation temperature. CONCLUSION: Changes in extraction pressure, temperature, time, pulverized degree and separation pressure affect the extracting results remarkably. The two kinds of chemical constituents were separated by recrystallization from C. monieri and identified by the methods of UV, IR, MS, NMR.