Phil-Sun Oh

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.

Allergy-related cytokines (IL-4 and TNF-α) are induced by Di(2-ethylhexyl) phthalate and attenuated by plant-originated glycoprotein (75 kDa) in HMC-1 cells

Phthalate esters as plasticizers have been widespread in the environment and may be associated with development of allergic diseases such as asthma and atopic dermatitis. In this study, we demonstrated that the CTB glycoprotein attenuates allergic reactions caused by di(2‐ethylhexyl) phthalate (DEHP) in human mast cells (HMC‐1). This experiment evaluated degranulation of histamine and β‐hexosaminidase as well as activities of protein kinase C (PKC), stress‐activated protein kinase/c‐Jun N‐terminal kinase (SAPK/JNK), activator protein (AP)‐1 and interleukin (IL)‐4 and tumor necrosis factor (TNF)‐α using immunoblotting and reverse transcription‐polymerase chain reaction (RT‐PCR). Our results revealed that the CTB glycoprotein in the presence of DEHP inhibits degranulation of mast cell, translocation of PKC from cytosol to membrane, and phosphorylation of SAPK/JNK in HMC ‐1 cells. We also found that the CTB glycoprotein (100 μg mL−1) has suppressive effects on transcriptional activation of AP‐1, and on the expression of IL‐4 and TNF‐α in DEHP‐treated HMC‐1 cells. We suggest that the CTB glycoprotein inhibits degranulation of mast cells and expressions of cytokines in HMC‐1 cells.

IgE, COX-2, and IL-4 are Expressed by DEHP through p38 MAPK and Suppressed by Plant Glycoprotein (75 kDa) in ICR Mice

The aim of the present study was to evaluate the inhibitory effect of glycoprotein isolated from Cudrania tricuspidata Bureau (CTB glycoprotein) on di(2-ethylhexyl) phthalate (DEHP)-induced allergic inflammatory response in mice. We evaluated the activity of β-hexosaminidase, expression of cyclooxygenase (COX)-2, p38 mitogen-activated protein kinase (MAPK), and activator protein (AP)-1, and production of immunoglobulin (Ig)E and interleukin (IL)-4 in DEHP-treated RBL-2H3 cells and ICR mice. Our results revealed that the CTB glycoprotein inhibited the activity of β-hexosaminidase and production of IgE and IL-4 in serum from DEHP-treated mice. We also found that the CTB glycoprotein reduced arachidonic acid release, COX-2 expression, and AP-1 transcriptional activation through p38 MAPK phosphorylation in DEHP-treated RBL-2H3 cells. The activation of AP-1 was completely blocked by treatment with p38 MAPK inhibitor (SKF86002). The results from these experiments indicate that CTB glycoprotein effectively protects against the allergic inflammation response, mainly through downregulation of MAPK/AP-1 in the mast cell degranulation stage. In conclusion, we suggest that the CTB glycoprotein may be one component of health supplements for the prevention of allergic inflammation.

Anti-inflammatory activity of ethanol extract from Geranium sibiricum Linne.

ETHNOPHARMACOLOGICAL RELEVANCE Geranium sibiricum (Geraniaceae) Linne (GSL) is used to heal various disorders of the diarrhea and the intestinal inflammation as an herbal agent in East Asia. AIMS OF THE STUDY The purpose of the present study is to determine whether the ethanol (EtOH) extract of GSL regulates the inflammatory reaction stimulated by phorbol-12-myristate 13-acetate plus calcium ionophore A23187 (PMACI) in human mast cells (HMC-1). MATERIALS AND METHODS Western blot was used for activation of mitogen activated protein kinase (MAPK), transcription factors, induced nitric oxide synthase (iNOS), and cyclooxygenase (COX)-2 proteins. EMSA was for DNA binding activity. RT-PCR was used for gene expression. RESULTS EtOH extract of GSL (EGS) inhibits the expression of extracellular signal-regulated kinase (ERK), one of a MAPK, nuclear transcription factors involving nuclear factor (NF)-kappaB and Activator protein (AP)-1, COX-2 and iNOS. The results indicated that EGS decreased gene expression of interleukin (IL)-1beta and COX-2 in PMACI stimulated HMC-1 cells. CONCLUSION Hence, we speculate that EGS can use as a potent anti-inflammatory agent for inflammatory allergic diseases.

Modulatory effects of phytoglycoprotein (75 kDa) on allergic inflammatory cytokines in Di(2‐ethylhexyl) phthalate (DEHP)‐stimulated RBL‐2H3 cells

This study investigated the inhibitory effect of a glycoprotein isolated from Cudrania tricuspidata Bureau (CTB glycoprotein) on di(2‐ethylhexyl) phthalate (DEHP)‐induced mast cell degranulation and related signaling cascade in RBL‐2H3 cells. This experiment evaluated the intracellular Ca2+ level, and the activities of protein kinase C (PKC), mitogen‐activated protein kinase (MAPK), transcription factor, and the cytokines in DEHP‐treated RBL‐2H3 cells. Our results revealed that the CTB glycoprotein in the presence of DEHP inhibits the release of histamine and expression of interleukin (IL)‐4, IL‐6, and TNF‐α in RBL‐2H3 cells. We also found that the CTB glycoprotein inhibits the intracellular Ca2+ level, translocation of PKC from cytosol to membrane and the phosphorylation of ERK1/2 in cells. Moreover, the CTB glycoprotein (100 µg/ml) has suppressive effects on transcriptional activation of nuclear factor (NF)‐κB in DEHP‐treated RBL‐2H3 cells. The activation of NF‐κB was collectively blocked by treatment with PKC inhibitor (staurosporine) as well as ERK1/2 inhibitor (PD98059), respectively. The results from these experiments indicated that the CTB glycoprotein inhibits release of histamine and expressions of IL‐4, IL‐6, and TNF‐α via down regulations of PKC/MAPK and NF‐κB on the stage of mast cell degranulation induced by DEHP. Moreover, oral administration of CTB glycoprotein (10–20 mg/kg) inhibited compound 48/80‐mediated systemic reaction in mice. In conclusion, we speculated that the CTB glycoprotein might be one component for preparation of health supplements for prevention of allergic immune disorders. J. Cell. Biochem. 109: 124–131, 2010.

Blocking of intracellular ROS production by phytoglycoprotein (30 kDa) causes anti-proliferation in bisphenol A-stimulated Chang liver cells

Dioscorea batatas Decne (DBD) is traditionally used to heal inflammatory disease as a folk medicine. It was reported that a glycoprotein (DBD glycoprotein) with a molecular weight of 30 kDa was isolated from DBD and consists of carbohydrate (83.75%) and protein (16.25%) moieties. The previous results showed that it has a strong scavenging activity against hydroxyl radicals without any pro‐oxidant activity in the cell‐free system. The purpose of the present study was to show whether or not the DBD glycoprotein inhibits cell proliferation‐related signal transduction stimulated by bisphenol A (BPA, an environmental hormone) in Chang liver cells. The results in this study indicated that DBD glycoprotein (200 µg ml−1) has suppressive effects on abnormal cell viability, production of intracellular reactive oxygen species (ROS) and nitric oxide (NO) in BPA (50 µm)‐induced Chang liver cells by blocking the phosphorylation of mitogen‐activated protein kinase (MAPK) and activating protein‐1 (AP‐1) activity. In addition, DBD glycoprotein (200 µg ml−1) normalized the activity of catalase (CAT) and glutathione peroxidase (GPx). Consequently, DBD glycoprotein inhibits the expression of proliferating cell nuclear antigen (PCNA, cell proliferation maker) stimulated by BPA. Therefore, it is speculated that DBD glycoprotein protects against carcinogenic events caused by BPA in Chang liver cells. Copyright

Protective activity of 30kDa phytoglycoprotein from glucose/glucose oxidase-induced cell death in primary cultured mouse thymocytes.

Dioscorea batatas Decne (DBD) has been traditionally used as herbal agent in folk medicine. DBD glycoprotein with a molecular weight of 30kDa consists of carbohydrate (83.75%) and protein (16.25%), and has a strong anti-oxidative activity. To understand the protection from thymocytes death, we evaluated the activity changes of pro-apoptotic factors [cytochrome c, caspase 3, poly(ADP-ribose) polymerase (PARP), AP-1, NF-κB and nitric oxide (NO)] by DBD glycoprotein from glucose/glucose oxidase (G/GO)-induced cell death in primary cultured mouse thymocytes. In the activity of the apoptotic related proteins [cytochrome c, caspase 3 and PARP], the results showed that DBD glycoprotein (200μg/ml) has an inhibitory effect on cytochrome c release into cytosol, caspase 3 activation and PARP cleavage in thymocytes. In the transcription factors (AP-1 and NF-κB) activity and NO production, the activities of NF-κB and NO production significantly decreased after DBD glycoprotein (200μg/ml) treatment for 4h in G/GO-induced thymocytes, compared with the control. Therefore, we speculate that DBD glycoprotein is one of the natural compounds for the protection of thymocytes that can produce cytokines.

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.

Glycoprotein isolated from Dioscorea batatas Decne modulates expressions of IL-4 and IL-10 in primary-cultured mouse lymphocytes.

In the present study, we investigated the anti‐allergic activity of a glycoprotein isolated from Dioscorea batatas Decne (DBD glycoprotein, 30 kDa) on ovalbumin (OVA, 100 µg ml−1)‐induced T helper (Th) type‐2 response in primary cultured mouse lymphocytes. Our results revealed that the DBD glycoprotein (200 µg ml−1) significantly attenuated the expressions of interleukin (IL)‐4 and IL‐10, whereas enhanced the expression of interferon (IFN)‐γ in OVA (100 µg ml−1)‐treated primary cultured lymphocytes. We also found that the DBD glycoprotein has inhibitory effects on phosphorylations of signal transducer and activator of transcription (STAT)‐6, p44/42 mitogen‐activated protein kinase (MAPK) and p38 MAPK in primary cultured lymphocytes. Interestingly, the DBD glycoprotein suppressed the transcriptional activation of GATA‐binding protein 3 (GATA‐3), whereas it enhanced the activity of t‐box expressed in T cells (T‐bet) in OVA‐stimulated lymphocytes. The results from these experiments indicated that the DBD glycoprotein inhibits expressions of IL‐4 and IL‐10 through modulation of GATA‐3, STAT‐6, p44/42 MAPK and p38 MAPK in mouse lymphocytes. Therefore, we speculated that the DBD glycoprotein might be one component for preparation of nutraceutical health supplements for prevention of Th2 cell response‐related immune disorder. Copyright