Sei-Jung Lee

Hypolipidemic and Antioxidative Effects of the Plant Glycoprotein (36 kDa) from Rhus verniciflua Stokes Fruit in Triton WR-1339-Induced Hyperlipidemic Mice

We investigated the hypolipidemic and antioxidative effects on male ICR mice of a glycoprotein isolated from Rhus verniciflua Stokes (RVS) fruit. The administration of the RVS glycoprotein (100 mg/kg) for two weeks resulted in a significant decrease in such plasma lipid levels as total cholesterol (TC), triglyceride (TG), and low-density lipoprotein (LDL). The levels of TC, TG and LDL in the hyperlipidemic model were significantly increased, whereas the high-density lipoprotein (HDL) level was considerably decreased. The 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase activity and the level of thiobarbituric acid-reactive substances (TBARS) were significantly elevated, whereas the production of nitric oxide (NO) was diminished. Moreover, the administration of the RVS glycoprotein prior to inducing hyperlipidemic mice suppressed the increase in the plasma lipid levels (TC, TG and LDL), and decrease in the HDL level in Triton WR-1339-induced hyperlipidemic mice. Furthermore, the RVS glycoprotein significantly inhibited the activity of HMG-CoA reductase and the levels of TBARS in the hyperlipidemic mice. In addition, the activities of detoxicant enzymes [catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx)] were gradually augmented after a supplement with the RVS glycoprotein. The results suggest that the RVS glycoprotein would be effective in preventing an increase in the plasma lipid levels and in improving the antioxidant levels. This protein might be useful as a therapeutic agent.

Hepatoprotective and hypolipidaemic effects of glycoprotein isolated from Gardenia jasminoides ellis in mice.

1 The present study was performed to investigate the hepatoprotective and hypolipidaemic effects of a 27 kDa glycoprotein isolated from Gardenia jasminoides Ellis (GJE glycoprotein) in glucose/glucose oxidase (G/GO)‐treated BNL CL.2 cells, as well as in CCl4, Triton WR‐1339 and corn oil‐treated mice. 2 In G/GO‐treated BNL CL.2 cells, the results showed that GJE glycoprotein has an inhibitory effect on G/GO‐induced cytotoxicity and intracellular reactive oxygen species production. In addition, GJE glycoprotein has an anti‐oxidant effect against the lipid peroxidation process in the Fe2+/ascorbic acid system. 3 In CCl4 (1.0 mL/kg)‐treated mice, pretreatment with GJE glycoprotein (80 mg/kg) blocked lactate dehydrogenase release and the formation of thiobarbituric acid‐reactive substances. In addition, in these mice GJE resulted in increased nitric oxide production and the activation of anti‐oxidant enzymes, accompanied by the inhibition of the cytotoxic‐related signals hepatic cytochrome c, nuclear factor‐κB and activator protein‐1. 4 In both Triton WR‐1339 (400 mg/kg) and corn oil (1.0 g/kg)‐treated mice, pretreatment with GJE glycoprotein (80 mg/kg) lowered the levels of plasma lipoproteins (triglyceride, total cholesterol and low‐density lipoprotein). 5 On the basis of these results, we assume that GJE glycoprotein can ameliorate liver function, because it has hepatoprotective and hypolipidaemic activities.

Protective effect of glycoprotein isolated from Ulmus davidiana Nakai on carbon tetrachloride‐induced mouse liver injury

This study was carried out to evaluate the hepatoprotective activity of glycoprotein isolated from the stems of Ulmus davidiana Nakai (UDN), which has been used as an anti‐inflammatory agent in folk medicine. We evaluated lipid peroxidation in glucose/glucose oxidase (G/GO)‐induced BNL CL.2 cells and measured thiobarbituric acid reactive substances (TBARS), lactate dehydrogenase (LDH), nitric oxide (NO), antioxidant enzyme (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)), activity of cytotoxic‐related signals (hepatic cytochrome c, nuclear factor‐kappa B (NF‐κB) and activator protein‐1 (AP‐1)) and levels of plasma lipids (triglyceride (TG) and total cholesterol (TC)) in carbon tetrachloride (CCl4, 1.0 mL kg−1)‐induced A/J mouse. The results in G/GO‐induced BNL CL.2 cells showed that UDN glycoprotein had a dose‐dependent inhibitory effect on lipid peroxidation. The results in carbon tetrachloride (CCl4, 1.0 mL kg−1)‐induced A/J mouse indicated that treatment with UDN glycoprotein (40 mg kg−1) lowered LDH activity and TBARS formation, and increased NO production and antioxidant enzymes activity, compared with control. Also, our finding from CCl4‐treated mice after pretreatment with UDN glycoprotein demonstrated that the activity of cytotoxic‐related signals decreased but the levels of plasma lipids increased, compared with CCl4 treatment alone. Here, we speculate that UDN glycoprotein has a protective character to CCl4‐induced mouse liver injury.

Glycoprotein isolated from Gardenia jasminoides Ellis has a scavenging activity against oxygen radicals and inhibits the oxygen radical-induced protein kinase C alpha and nuclear factor-kappa B in NIH/3T3 cells.

This study was earned out to investigate the antioxidative and anti-apoptotic effects of glycoprotein isolated from Gardenia jasminoides Ellis fruit (GJE glycoprotein), which has been used to heal hepatic and inflammatory diseases in folk medicine. GJE glycoprotein showed a single band with a molecular weight of 27kDa on the 15% sodium dodecyl sulfate polyacrylamide gel. It consists of a carbohydrate component (57.65%) and a protein component (42.35%). GJE glycoprotein has dose-dependent scavenging activities for DPPH, lipid peroxyl, superoxide anion and hydroxyl radicals in cell-free systems. We also evaluated the protective and anti-apoptotic activities of GJE glycoprotein on the glucose/glucose oxidase (G/GO)-induced or hypoxanthine/xanthine oxidase (HX/XO)-induced cytotoxicity and apoptosis systems in NIH/3T3 cells, using 3-(4,5-diinettiylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT), DNA fragmentation and H33342/ethidium bromide staining assays, respectively. Results in this experiment showed that GJE glycoprotein has dose-dependent blocking activities against G/GO- or HX/XO-induced cytotoxicity and apoptosis. In addition, we investigated whether GJE glycoprotein blocks the activation of redox-sensitive signal mediators, protein kinase C alpha (PKCα) and nuclear factor-kappa B (NF-κB) in G/GO or HX/XO-induced apoptotic NIH/3T3 cells, using a Western blot analysis and an electrophoretic mobility shift assay (EMSA). We found that 100μg/ml GJE glycoprotein has an inhibitory effect on PKCα translocation and the DNA binding activity of (NF-κB). Here, we speculate that GJE glycoprotein is a natural antioxidant and one of the modulators of apoptotic signal pathways in NIH/3T3 cells.

Rhus verniciflua Stokes glycoprotein (36kDa) has protective activity on carbon tetrachloride-induced liver injury in mice.

This study was carried out to investigate the hepatoprotective effect of the glycoprotein isolated from Rhus verniciflua Stokes (RVS), which has traditionally been used for healing of inflammatory diseases. We evaluated the activities of alanine aminotransferase (ALT), lactate dehydrogenase (LDH), thiobarbituric acid-reactive substances (TBARS), and antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx)] activities in treatment with carbon tetrachloride (CCl(4)) in vivo. When mice were treated with CCl(4) in the absence of RVS glycoprotein, the activities of ALT, LDH, and TBARS were increased, while the antioxidant enzymes activities were decreased. However, when the mice were treated with CCl(4) in the presence of RVS glycoprotein, the activities of ALT, LDH, and TBARS were significantly reduced and SOD, CAT, and GPx activities were remarkably increased. In addition, RVS glycoprotein increased the nitric oxide (NO) production and decreased the nuclear factor-kappa B (NF-κB) and activator protein-1 (AP-1) activation in CCl(4)-treated mice. Collectively, these results pointed out that RVS glycoprotein can inhibit lipid peroxidation, enhance the activities of antioxidant enzymes, increase the NO production, and decrease the NF-κB and AP-1 activations. Therefore, we speculate that RVS glycoprotein protects from liver damage through its radical scavenging ability.

ZPDC glycoprotein (24 kDa) induces apoptosis and enhances activity of NK cells in N-nitrosodiethylamine-injected Balb/c

Natural killer (NK) cells have anti-tumor activity in hepatocellular carcinoma (HCC) using secreting granules and cytotoxic ability. Recently, we isolated glycoprotein from Zanthoxylum piperitum DC (ZPDC) has anti-oxidant effect and anti-cancer effect. The objective of this study was to determine whether ZPDC glycoprotein enhances activity of NK cells and induces apoptosis of liver cancer cells in diethylnitrosamine (DEN)-treated Balb/c mice. This study evaluated the secreting of perforin and granzyme B and cytotoxicity of NK cells, interleukin (IL)-2 and IL-12, apoptosis-related factors (bid, cytochrome c, and caspase-3) in liver tissue using Immunoblot and ELISA. The results demonstrated that ZPDC glycoprotein (20mg/kg, BW) induces secretion of perforin and granzyme B and NK cells activity. Also, it induces expression of apoptosis-related factors (bid, cytochrome c, and caspase-3) in liver tissues. Collectively, ZPDC glycoprotein may have potential applications to prevent hepatocarcinogenesis without immunosuppression.

A 116-kDa phytoglycoprotein inhibits aberrant crypt foci formation through modulation of manganese superoxide dismutase, inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-kappa B, activator protein-1, and proliferating cell nuclear antigen in 1,2-dimethylhydrazine/dextran sodium sulfate-treated ICR mice.

The 116-kDa Ulmus davidiana Nakai (UDN) glycoprotein is a naturally occurring phytoglycoprotein found in the stem of UDN. In this study, we investigated the chemopreventive effect of UDN glycoprotein on inflammation-mediated colorectal carcinogenesis induced by 10 mg/kg 1,2-dimethylhydrazine and 2% dextran sodium sulfate in ICR mice. Consumption of UDN glycoprotein (0.01 and 0.02%) significantly reduced the frequency of colonic aberrant crypt foci, the expression of colonic proliferating cell nuclear antigen, and the release of plasma lactate dehydrogenase without any cytotoxic activity at the initiation stage of colorectal carcinogenesis in 1,2-dimethylhydrazine/dextran sodium sulfate-treated mice. In addition, UDN glycoprotein has antioxidative effects on the formation of plasma thiobarbituric acid reactive substances and on the production of plasma inducible nitric oxide, accompanying the normalizing effects on the activity of colonic antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) in the mice. UDN glycoprotein intake also remarkably attenuated the expression of inflammation-related factors (inducible nitric oxide synthase and cyclooxygenase-2) and the DNA-binding activity of redox-sensitive transcription factors (nuclear factor-kappa B and activator protein-1) in the mice. Collectively, the results suggest that UDN glycoprotein has chemopreventive potential at the initiation stage of colorectal cancer by reducing the factors responsible for oxidative stress, inflammation, and carcinogenesis.

Preventive effects of ZPDC glycoprotein (24 kDa) on hepatotoxicity induced by mercury chloride in vitro and in vivo

Mercury is a potent environmental contaminant that exerts toxic effect on various vital organs in the human body. Recently, we isolated glycoprotein from Zanthoxylum piperitum DC (ZPDC), which has antioxidant and anticancer effects. In the present study, we determined the preventive effects of ZPDC glycoprotein on hepatic damage induced by mercury chloride (HgCl2). We evaluated the activities of lactate dehydrogenase (LDH), alanine aminotransferase (ALT), antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], extracellular signal‐regulated kinase (ERK)1/2, p38 mitogen‐activated protein kinase (MAPK), cyclo‐oxygenase (COX‐2), inducible nitric oxide synthetase (iNOS), and activator protein (AP‐1) and the quantitative expressions of nuclear factor E2‐related factor (Nrf2), heme oxygenase (HO‐1), metallothionein (MT) and reduced glutathione (GSH) in mercury‐chloride‐exposed (50 μM and 10 mg/kg body weight) primary cultured hepatocytes and ICR mice, using biochemical assays, radioactivity and immunoblot analysis. The results demonstrated that ZPDC glycoprotein decreased the levels of LDH, ALT, HO‐1 and MT, whereas it increased the activities of hepatic antioxidant enzymes (SOD, CAT and GPx) and reduced GSH in mercury‐chloride‐exposed primary cultured hepatocytes. Also, it suppressed arachidonic acid release and expression of ERK, p38 MAPK, COX‐2, iNOS, AP‐1 and Nrf‐2 in primary cultured hepatocytes and ICR mice exposed to mercury chloride. Collectively, ZPDC glycoprotein may have potential applications to prevent hepatotoxicity induced by mercury chloride. Copyright

ZPDC glycoprotein inhibits inflammation-related cytokine and protein via nuclear factor-kappa B in dextran sulfate sodium-stimulated ICR mouse

The purpose of this study is to investigate the anti-inflammatory potentials of a 24-kDa glycoprotein isolated from Zanthoxylum piperitum DC fruit (ZPDC glycoprotein) in dextran sulfate sodium (DSS)-stimulated ICR mouse colitis. ZPDC glycoprotein was administered to mice at 10 and 20 mg/kg for 7 days and then the mice were co-treated with 5% DSS for another 7 days in presence of ZPDC glycoprotein and killed on day 15. The results showed that ZPDC glycoprotein has inhibitory effects on levels of disease activity index and large intestine shortening in DSS-treated mice. In addition, ZPDC glycoprotein suppresses the formation of thiobarbituric acid reactive substances, production of inducible nitric oxide, and release of lactate dehydrogenase in DSS-treated mice plasma. Interestingly, we found that consumption of ZPDC glycoprotein (20 mg/kg) significantly inhibited the expressions of interleukin-1beta and tumor necrosis factor-alpha, inducible nitric oxide synthase, and cyclooxygenase-2 via modulation of transcriptional activity of nuclear factor-kappa B in DSS-treated mice colon. Collectively, these results suggest that ZPDC glycoprotein is useful for prevention of inflammatory gastrointestinal diseases.

Phytoglycoprotein (24 kDa) inhibits expression of PCNA via PKCα and MAPKs in oxygen radical-stimulated Chang liver cells

The purpose of this study was to investigate the inhibitory effect of 24-kDa glycoprotein isolated from Zanthoxylum piperitum DC fruit (ZPDC glycoprotein) on glucose/glucose oxidase (G/GO)- or hypoxanthine/xanthine oxidase (HX/XO)-induced cell proliferation in Chang liver cells. We found that ZPDC glycoprotein has significant scavenging effect on the production of intracellular H2O2 without cytotoxicity in G/GO- or HX/XO-treated in Chang liver cells. In the G/GO or HX/XO-stimulated protein kinases activity, ZPDC glycoprotein inhibited translocation of protein kinase C alpha (PKCalpha) to membrane and phosphorylation of extracellular signal-regulated kinase, p38 MAP kinase and c-Jun N-terminal kinase, respectively. In the G/GO or HX/XO-stimulated transcriptional activity, ZPDC glycoprotein also blocked the DNA binding activities of nuclear factor-kappa B and activator protein-1 and attenuated the activities of p50, p65, c-Jun and c-Fos, respectively. Finally, in the G/GO or HX/XO-stimulated cell proliferation, the activity of proliferating cell nuclear antigen was significantly blocked by treatment with ZPDC glycoprotein as well as protein kinase C inhibitor and mitogen-activated protein kinase inhibitors. On the basis of these results, we speculate that this glycoprotein is one of the natural antioxidants and of the modulators on abnormal activation of cell proliferation-related molecules in Chang liver cells.