Chin-Lin Hsu

Effects of Polyphenolic Compounds on Tumor Necrosis Factor-α (TNF-α)-Induced Changes of Adipokines and Oxidative Stress in 3T3-L1 Adipocytes

Over the last few decades, obesity has become a global epidemic in both developed and developing countries. Recent studies have indicated that obesity is closely associated with chronic inflammation characterized by abnormal levels of adipocytokines and inflammatory cytokines in adipocytes. The aim of this work was to study the effects of 21 polyphenolic compounds on tumor necrosis factor-α (TNF-α)-induced changes of adipokines and oxidative stress in 3T3-L1 adipocytes. The results showed that p-coumaric acid, quercetin, and resveratrol have greater inhibition (p < 0.05) of a TNF-α-induced increase in the production of interleukin-6 (IL-6) among 21 tested polyphenolic compounds. p-Coumaric acid, quercetin, and resveratrol demonstrated inhibitions of TNF-α-induced changes in levels of monocyte chemoattractant protein-1 (MCP-1), plasminogen activator inhibitor-1 (PAI-1), and intracellular reactive oxygen species (ROS) in 3T3-L1 adipocytes. Furthermore, p-coumaric acid, quercetin, and resveratrol increased levels (p < 0.05) of secreted adiponectin, superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GPx), and glutathione S-transferase (GST) in TNF-α-treated 3T3-L1 adipocytes. These results indicate that the inhibition of TNF-α-induced changes of adipokines and oxidative stress by some polyphenolic compounds might have further implications in preventing obesity-related pathologies.

Antiobesity and Hypolipidemic Effects of Polyphenol-Rich Longan (Dimocarpus longans Lour.) Flower Water Extract in Hypercaloric-Dietary Rats

Plenty of polyphenols, i.e. phenolic acids and flavonoids, were found in longan flower water extract (LFWE) through spectrophotometric and HPLC analyses. Antiobesity and hypolipidemic effects of polyphenol-rich longan flower water extract (LFWE) were investigated in this study. Eight male rats per group were assigned randomly to one of the following dietary groups: (1) normal-caloric diet and pure water (NCD + NDW); (2) hypercaloric diet and pure water (HCD + NDW); (3) HCD and 1.25% (w/v) LFWE (HCD + 1.25% LFWE); (4) HCD and 2.5% (w/v) LFWE (HCD + 2.5% LFWE) for 9 weeks. Body weight, size of epididymal fat, serum triglyceride level and atherogenic index, and hepatic lipids were decreased (p < 0.05) in HCD rats by drinking 2.5% LFWE which may result from downregulated (p < 0.05) pancreatic lipase activity, and sterol regulatory element binding protein-1c (SREBP-1c) and fatty acid synthase (FAS) gene expressions, as well as upregulated (p < 0.05) LDL receptor (LDLR) and peroxisome proliferator-activated-receptor-alpha (PPAR-alpha) gene expressions, and also increased (p < 0.05) fecal triglyceride excretions. Therefore, polyphenol-rich LFWE indeed characterizes antiobesity and hypolipidemic effects in vivo.

Antioxidant and anti-inflammatory effects of Orthosiphon aristatus and its bioactive compounds.

Orthosiphon aristatus (Blume) Miq., which can be used as a food ingredient, is grown throughout Southeast Asia and Australia. O. aristatus is frequently used for the treatment of renal inflammation, kidney stones and dysuria. The focus of the current work was to study the antioxidant and anti-inflammatory effects of methanol, ethanol and water extracts from O. aristatus (abbreviated as MEOA, EEOA and WEOA, respectively). The evaluation of antioxidant activity was determined by total phenolics, Trolox equivalent antioxidant capacity (TEAC), oxygen-radical absorbance capacity (ORAC) and cellular antioxidant activity (CAA) assays. These assays demonstrated a relatively high antioxidant activity for MEOA and EEOA. These results revealed that EEOA had the most prominent inhibitory effect on lipopolysaccharide (LPS)-stimulated nitric oxide (NO), prostaglandin E(2) (PGE(2)) and intracellular reactive oxygen species (ROS) production in RAW 264.7 cells. A high performance liquid chromatography profile indicated that MEOA and EEOA contained both ursolic acid and oleanolic acid. Moreover, ursolic acid significantly reduced NO production in LPS-stimulated RAW 264.7 cells. Both EEOA and ursolic acid inhibited LPS-stimulated protein and mRNA expression of both inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in these cells. These results demonstrate that EEOA and its bioactive compound, ursolic acid, suppress LPS-induced NO and PGE(2) production by inhibiting ROS generation, along with reducing expression of iNOS and COX-2 in RAW 264.7 cells.

Anti-inflammatory effects of the roots of Alpinia pricei Hayata and its phenolic compounds.

Alpinia pricei Hayata is cultivated throughout Asia and is an endemic plant in Taiwan. The leaf and root of this plant are used for traditional wrapping of food and as a cooking substitute for fresh ginger. The aim of this work was to study the in vitro anti-inflammatory effects of ethanol extracts from A. pricei Hayata (EEAP) and its phenolic compounds. High-performance liquid chromatography (HPLC) profiling indicated that EEAP contained caffeic acid, chlorogenic acid, ferulic acid, p-hydroxybenzoic acid, rutin, apigenin, curcumin and pinocembrin. EEAP and its phenolic compounds, apigenin, curcumin, and pinocembrin, inhibited lipopolysaccharide (LPS)-stimulated nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in RAW 264.7 cells. Furthermore, EEAP, apigenin, curcumin, and pinocembrin decreased LPS-mediated induction of protein and mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in RAW 264.7 cells. In addition, EEAP and its major active compound pinocembrin inhibited LPS-induced nuclear translocation of nuclear factor-kappaB (NF-kappaB) and NF-kappaB-mediated reporter gene expression. EEAP and pinocembrin also significantly inhibited LPS-induced intracellular reactive oxygen species (ROS) production in RAW 264.7 cells. When these results are taken together, they indicate that EEAP and pinocembrin suppressed LPS-induced NO and PGE(2) production by inhibition of NF-kappaB nuclear translocation and ROS generation.

Lucidenic Acid B Induces Apoptosis in Human Leukemia Cells via a Mitochondria-Mediated Pathway

Ganoderma lucidum is known as a medicinal mushroom used in traditional Chinese medicine. In the present study, the effect of lucidenic acids (A, B, C, and N) isolated from a new G. lucidum (YK-02) on induction of cell apoptosis and the apoptotic pathway in HL-60 cells were investigated. The results demonstrated that lucidenic acids decreased cell population growth of HL-60 cells, assessed with the MTT assay. The cell cycle assay indicated that treatment of HL-60 cells with lucidenic acid A, C, and N caused cell cycle arrest in the G 1 phase. Lucidenic acid B (LAB) did not affect the cell cycle profile; however, it increased the number of early and late apoptotic cells but not necrotic cells. Treatment of HL-60 cells with LAB caused loss of mitochondria membrane potential. Moreover, the ratio of expression levels of pro- and antiapoptotic Bcl-2 family members was changed by LAB treatment. LAB-induced apoptosis involved release of mitochondria cytochrome c and subsequently induced the activation of caspase-9 and caspase-3, which were followed by cleavage of poly(ADP-ribose) polymerase (PARP). Pretreatment with a general caspase-9 inhibitor (Z-LEHD-FMK) and caspase-3 inhibitor (Z-DEVD-FMK) prevented LAB from inhibiting cell viability in HL-60 cells. Our finding may be critical to the chemopreventive potential of lucidenic acid B.

Hepatoprotection of noni juice against chronic alcohol consumption: lipid homeostasis, antioxidation, alcohol clearance, and anti-inflammation.

Chronic alcohol consumption leads to steatohepatitis and cirrhosis. Naturally fermented noni juice (NJ) contains polyphenols, polysaccharides, and some trace minerals. This study explored protective effects of NJ against chronic alcohol consumption. Mice were assigned randomly to one of the following groups: (1) control, control liquid diet and distilled water; (2) alcohol, alcohol liquid diet and distilled water; (3) Alc+NJ_1X, alcohol liquid diet and 5 mL NJ/kg BW; (4) Alc+NJ_2X, alcohol liquid diet and 10 mL NJ/kg BW; (5) Alc+NJ_3X, alcohol and 15 mL NJ/kg BW for 4 weeks. NJ decreased (p < 0.05) serum AST, ALT, and alcohol levels and liver lipids, as well as increased (p < 0.05) daily fecal lipid outputs in alcohol-diet fed mice. NJ supplementation not only down-regulated (p < 0.05) lipogenesis but also up-regulated (p < 0.05) fatty acid β-oxidation in livers of alcohol-diet fed mice. NJ also accelerated alcohol clearance via increased (p < 0.05) hepatic ADH and ALDH activities. NJ increased (p < 0.05) hepatic TEAC and GSH levels but decreased (p < 0.05) TBARS value and TLR2/4, P38, ERK 1/2, NFκB P65, iNOS, COX-2, TNF-α, and IL-1β expressions in alcohol-diet fed mice. NJ promotes hepatoprotection against alcohol-induced injury due to regulations of lipid homeostasis, antioxidant status, alcohol metabolism, and anti-inflammatory responses.

Anti-inflammatory effects of phenolic compounds isolated from the flowers of Nymphaea mexicana Zucc.

Nymphaea mexicana Zucc. is an aquatic plant species which belongs to the family Nymphaea and is commonly known as the yellow water lily. The aim of this work was to study the in vitro antiinflammatory effects of phenolic compounds isolated from the flowers of Nymphaea mexicana Zucc. Seven phenolic compounds including vanillic acid, 4-methoxy-3,5-dihydroxybenzoic acid, (2R,3R)-3,7-dihydroxyflavanone, naringenin (4), kaempferol 3-O-(3-O-acetyl-a-L-rhamnopyranoside), kaempferol 3-O-(2-O-acetyl-a-L-rhamnopyranoside), and quercetin 3-(30 0-acetylrhamnoside) (7) were isolated from the flowers of Nymphaea mexicana Zucc. These results revealed that compound 4 has the most prominent inhibitory effect on the LPS-stimulated nitric oxide (NO), monocyte chemotactic protein-1 (MCP-1), and tumor necrosis factor-alpha (TNF-a) production in RAW 264.7 macrophages. In addition, compound 4 also inhibited LPS-mediated induction of protein expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), and phospho-ERK in RAW 264.7 macrophages. Thus, compound 4 from the flowers of Nymphaea mexicana Zucc. may provide a potential therapeutic approach for inflammation-associated disorders.

Anticancer effects of Alpinia pricei Hayata roots.

The leaves and roots of Alpinia pricei Hayata are used as a traditional wrapping for food and as a cooking substitute for fresh ginger. Our previous study showed that ethanol extracts from the roots of A. pricei Hayata (EEAP) and its phenolic compounds have anti-inflammatory effects. The aims of this work were to further study the in vitro anticancer activity of EEAP and its active compounds with respect to various cancer cells. The results from an MTT assay demonstrated that EEAP decreased the cell population growth of CH27, HL-60, and A549 cells. Flow cytometric analysis of HL-60 cells exposed to EEAP showed that the number of apoptotic cells increased in a time- and dose-dependent manner. Western blot data revealed that EEAP stimulated an increase in the level of protein expression of Fas, FasL, caspase-8, and tBid. Moreover, the ratio of the expression levels of pro- and anti-apoptotic Bcl-2 family members was changed after treatment with EEAP. EEAP-induced apoptosis involved the release of mitochondrial cytochrome c and subsequently induced the activation of caspase-9 and caspase-3, which were followed by the cleavage of poly(ADP-ribose) polymerase (PARP). The results also demonstrated that phenolic compounds (caffeic acid, apigenin, curcumin, and pinocembrin) from EEAP decreased the rate of population growth of HL-60 cells. Treatment of HL-60 cells with these phenolic compounds caused the loss of mitochondrial membrane potential. Our finding could provide critical information regarding the chemopreventive potential of ethanol extracts from A. pricei Hayata. These results also demonstrate that the EEAP-induced apoptotic ability in HL-60 cells might be related to the phenolic compounds.

Anticancer Effects of Alpinia pricei Hayata Roots

The leaves and roots of Alpinia pricei Hayata are used as a traditional wrapping for food and as a cooking substitute for fresh ginger. Our previous study showed that ethanol extracts from the roots of A. pricei Hayata (EEAP) and its phenolic compounds have anti-inflammatory effects. The aims of this work were to further study the in vitro anticancer activity of EEAP and its active compounds with respect to various cancer cells. The results from an MTT assay demonstrated that EEAP decreased the cell population growth of CH27, HL-60, and A549 cells. Flow cytometric analysis of HL-60 cells exposed to EEAP showed that the number of apoptotic cells increased in a time- and dose-dependent manner. Western blot data revealed that EEAP stimulated an increase in the level of protein expression of Fas, FasL, caspase-8, and tBid. Moreover, the ratio of the expression levels of pro- and anti-apoptotic Bcl-2 family members was changed after treatment with EEAP. EEAP-induced apoptosis involved the release of mitochondrial cytochrome c and subsequently induced the activation of caspase-9 and caspase-3, which were followed by the cleavage of poly(ADP-ribose) polymerase (PARP). The results also demonstrated that phenolic compounds (caffeic acid, apigenin, curcumin, and pinocembrin) from EEAP decreased the rate of population growth of HL-60 cells. Treatment of HL-60 cells with these phenolic compounds caused the loss of mitochondrial membrane potential. Our finding could provide critical information regarding the chemopreventive potential of ethanol extracts from A. pricei Hayata. These results also demonstrate that the EEAP-induced apoptotic ability in HL-60 cells might be related to the phenolic compounds.

Alleviative effects of deep-seawater drinking water on hepatic lipid accumulation and oxidation induced by a high-fat diet

Background: Hepatic steatosis is defined as excessive amounts of triglyceride and other fats inside liver cells and has become an emergent liver disease in developed and developing countries. Methods: Deep seawater (DSW)300, DSW900, and DSW1500 drinking waters were formulated via a combination of reverse osmosis and electrodialysis. Hamsters on a high‐fat diet were assigned to drink the following solutions: (1) normal distilled water, (2) DSW300, (3) DSW900, or (4) DSW1500. Serum, liver, and fecal biochemical values, expression of hepatic genes related to fatty‐acid homeostasis, as well as liver antioxidative levels were measured after a 6‐week feeding period. Additionally, hematoxylin and eosin staining was used to investigate the liver histopathology. Results: Serum/liver lipids, liver sizes, liver malondialdehyde content, and serum aspartate aminotransferase and alanine aminotransferase of high‐fat diet hamsters were reduced (p < 0.05) by drinking DSW, while daily fecal lipid and bile acid outputs were increased (p < 0.05). DSW drinking water maintained (p < 0.05) higher liver glutathione and Trolox equivalent antioxidant capacity levels. Although hepatic sterol regulatory element‐binding protein‐1c, acetyl‐CoA carboxylase, fatty acid synthase, and malic enzyme gene expression were not (p > 0.05) altered, DSW drinking water upregulated (p < 0.05) hepatic peroxisome proliferator‐activated receptor‐alpha, retinoid X receptor alpha, and uncoupling protein‐2 gene expression in high‐fat diet hamsters. The lipid droplets in livers were also reduced in DSW‐drinking‐water groups as compared to those only drinking distilled water. Conclusion: DSW shows a preventive effect on development of hepatosteatosis induced by a high‐fat diet.