• Title/Summary/Keyword: GalN

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Galectin-1 from redlip mullet Liza haematocheilia: identification, immune responses, and functional characterization as pattern recognition receptors (PRRs) in host immune defense system

  • Chaehyeon Lim;Hyukjae Kwon;Jehee Lee
    • Fisheries and Aquatic Sciences
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    • v.25 no.11
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    • pp.559-571
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    • 2022
  • Galectins, a family of ß-galactoside-binding lectins, have emerged as soluble mediators in infected cells and pattern recognition receptors (PRRs) responsible for evoking and regulating innate immunity. The present study aimed to evaluate the role of galectin-1 in the host immune response of redlip mullet (Liza haematocheilia). We established a cDNA database for redlip mullet, and the cDNA sequence of galectin-1 (LhGal-1) was characterized. In silico analysis was performed, and the spatial and temporal expression patterns in gills and blood in response to lipopolysaccharide polyinosinic:polycytidylic acid, and Lactococcus garvieae were estimated via quantitative real-time PCR. Functional assays were conducted using recombinant protein to investigate carbohydrate binding, bacterial binding, and bacterial agglutination activity. LhGal-1 was composed of 135 amino acids. Conserved motifs (H-NPR, -N- and -W-E-R) within the carbohydrate recognition domain were found in LhGal-1. The tissue distribution revealed that the healthy stomach expressed high levels of LhGal-1. The temporal monitoring of LhGal-1 mRNA expression in the gill and blood showed its significant upregulation in response to immune challenges with different stimulants. rLhGal-1 exhibited binding activity in response to carbohydrates and bacteria. Moreover, the agglutination of rLhGal-1 against Escherichia coli was observed. Collectively, our findings suggest that LhGal-1 may function as a PRR in redlip mullet. Furthermore, LhGal-1 can be considered a significant gene to play a protective role in redlip mullet immune system.

Expression of the FLP recombinase of the 2 $\mu$m plasmid of yeast in the cultured cells of Bombyx mori using a transient expression vector (Yeast 2 $\mu$m 플라스미드 유래 FLP recombinase 유전자의 곤충 배양세포내 발현)

  • 강석우;윤은영
    • Journal of Sericultural and Entomological Science
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    • v.39 no.1
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    • pp.36-43
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    • 1997
  • In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

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Hepatoprotective Activities of Gomisin A and Gomisin N (Gomisin A 및 Gomisin N의 간독성 보호작용)

  • Heo Jeong-Haing;Park Jin-Gu;Cheon Ho-Jun;Kim Yeong-Shik;Kang Sam-Sik;Hung Tran Manh;Bae Ki-Hwan;Lee Sun-Mee
    • Korean Journal of Pharmacognosy
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    • v.37 no.4 s.147
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    • pp.294-301
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    • 2006
  • The aim of this study was to investigate the protective activity of gomisin A and gomisin N, bioactive lignan components isolated from Schizandae Fructus, on hepatocyte injury induced by carbon tetrachloride($CCl_4$, 10 mM), t-butyl hydroperoxide(TBH, 0.5 mM), and D-galactosamine(GalN, 30 mM). Primary cultures of rat hepatocyte(18 h culture) were treated with $CCl_4$, TBH or GalN and various concentrations(0.1, 1, 10, $100{\mu}M$) of gomisin A or gomisin N. $CCl_4$ significantly increased the levels of lactate dehydrogenase(LDH), alanine aminotransferase(ALT), and aspartate aminotransferase(AST). These increases were inhibited by gomisin N. TBH significantly increased the level of AST; an increase that was inhibited by gomisin N. GalN markedly increased the levels of LDH and ALT, and these increases was significantly inhibited by both gomisin A and gomisin N. These results suggest that gomisin A and gomisin N have the hepatoprotective activity.

Ginseng-derived type I rhamnogalacturonan polysaccharide binds to galectin-8 and antagonizes its function

  • Yi Zheng;Yunlong Si;Xuejiao Xu;Hongming Gu;Zhen He;Zihan Zhao;Zhangkai Feng;Jiyong Su;Kevin H. Mayo;Yifa Zhou;Guihua Tai
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.202-210
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    • 2024
  • Background: Panax ginseng Meyer polysaccharides exhibit various biological functions, like antagonizing galectin-3-mediated cell adhesion and migration. Galectin-8 (Gal-8), with its linker-joined N- and C-terminal carbohydrate recognition domains (CRDs), is also crucial to these biological processes, and thus plays a role in various pathological disorders. Yet the effect of ginseng-derived polysaccharides in modulating Gal-8 function has remained unclear. Methods: P. ginseng-derived pectin was chromatographically isolated and enzymatically digested to obtain a series of polysaccharides. Biolayer Interferometry (BLI) quantified their binding affinity to Gal-8, and their inhibitory effects on Gal-8 was assessed by hemagglutination, cell migration and T-cell apoptosis. Results: Our ginseng-derived pectin polysaccharides consist mostly of rhamnogalacturonan-I (RG-I) and homogalacturonan (HG). BLI shows that Gal-8 binding rests primarily in RG-I and its β-1,4-galactan side chains, with sub-micromolar KD values. Both N- and C-terminal Gal-8 CRDs bind RG-I, with binding correlated with Gal-8-mediated function. Conclusion: P. ginseng RG-I pectin β-1,4-galactan side chains are crucial to binding Gal-8 and antagonizing its function. This study enhances our understanding of galectin-sugar interactions, information that may be used in the development of pharmaceutical agents targeting Gal-8.

Effects of Panax ginseng on Galactosamine-induced Cytotoxicity in Primary Cultured Rat Hepatocytes (인삼 분획물이 Galactosamine에 의하여 손상된 일차배양한 흰쥐의 간세포에 미치는 영향)

  • Song, Jin-Ho;Park, Mi-Jung;Kim, Eun;Kim, Young-Choong
    • YAKHAK HOEJI
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    • v.34 no.5
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    • pp.341-347
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    • 1990
  • The anti-hepatotoxic activity of Panax ginseng was studied using galactosamine (GalN)-induced cytotoxicity in primary cultured rat hepatocytes. Panax ginseng was fractionated into dammarane glycosides and protein fractions. The dammarane glycosides was further fractionated into panaxadiol and panaxatriol glycosides fractions. The protein fraction was further fractionated into four groups according to the molecular weight; larger than 10,000 dalton, between 5,000 and 10,000 dalton, between 1,000 and 5,000 dalton and between 500 and 1,000 dalton. A significant lowering action on the elevated glutamicpyruvic transaminase (GPT) activity in the culture medium of hepatocytes treated with 1.5 mM GalN was noticed with all four protein fractions studied at the concentration of both $50\;{\mu}g/ml$ and $100\;{\mu}g/ml$. However, the effect of dammarane glycosides fractions was not significant. It was noted that the addition of $100\;{\mu}g/ml$ of protein fractions smaller than 5,000 dalton significantly enhanced the syntheses of protein and RNA in the damaged hepatocytes induced by the treatment of 1.5 mM GalN. Dammarane glycosides fractions significantly enhanced protein synthesis at the concentration of $100\;{\mu}g/ml$ in the damaged hepatocytes by treatment of 1.5 mM GalN.

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Hepatoprotective Effect of Ethanol Extract of xBrassicaraphanus on Liver Injury in Rats Treated by D-galactosamine (배무채 에탄올층의 D-galactosamine 간손상에 대한 보호효과)

  • Rhee, Yun-Hee;Lee, Eun-ok;Lee, Hyo-Jung;Shim, Bum-Sang;Ahn, Kyoo-Seok;Choi, Jong-Won;Lee, Soo-Seong;Yoon, Byong-Su;Kim, Sung-Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.6
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    • pp.1388-1393
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    • 2007
  • The protective effect of xBrassicoraphanus (BR) on liver inhury was evaluated in the rats with liver injury induced by i.p. injection of D-galactosamine (GalN) following 2 week oral treatment of ethanol extract of xBrassicoraphanus (EBR). EBR (200 mg/kg) significantly suppressed the levels of ALT, AST, SDH, ${\gamma}-GT$, ALP, LDH and lipid peroxidation compared with GalN treated control, while EBR at 100 mg/kg significantly suppressed AST and ${\gamma}-GT$. Similarly, EBR at 200 mg/kg significantly attenuated the levels of Phase I enzymes such as XO, AO, AH and AD as well as significantly increased the levels of Phase II enzymes such as SOD, catalase and GSH-Px in the GalN treated rats. Taken together, these results indicate that the ethanol extract of xBrassicoraphanus may have a hepatoprotective effect against GalN induced liver injury, suggesting the ethanol extract of xBrassicoraphanus can be applied as hepatoprotective functional food. However, its mechanism should be further studied in molecular and cellular view points.

Acanthopanax koreanum Nakai modulates the immune response by inhibiting TLR 4-dependent cytokine production in rat model of endotoxic shock

  • Jung, Myung-Gi;Do, Gyeong-Min;Shin, Jae-Ho;Ham, Young Min;Park, Soo-Yeong;Kwon, Oran
    • Nutrition Research and Practice
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    • v.7 no.6
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    • pp.460-465
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    • 2013
  • The hepatoprotective activity of Acanthopanax koreanum Nakai extract (AE) was investigated against D-Galactosamine/Lipopolysaccharide (D-GalN/LPS)-induced liver failure rats compared with that of acanthoic acid (AA) isolated from AE. Although D-GalN/LPS (250 mg/kg body weight/$10{\mu}g/kg$ body weight, i.p.) induced hepatic damage, pretreatments with AE (1 and 3% AE/g day) and AA (0.037% AA, equivalent to 3% AE/g day) alleviated the hepatic damage. This effect was the result of a significant decrease in the activity of alanine transaminase. Concomitantly, both the nitric oxide and IL-6 levels in the plasma were significantly decreased by high-dose AE (AE3) treatment compared to the GalN/LPS control (AE0). This response resulted from the regulation of pro-inflammatory signaling via a decrease in TLR4 and CD14 mRNA levels in the liver. While a high degree of necrosis and hemorrhage were observed in the AE0, pretreatment with AE3 and AA reduced the extent of hepatocyte degeneration, necrosis, hemorrhage and inflammatory cell infiltrates compared to the AE0. In conclusion, these results suggest that especially high-dose AE are capable of alleviating D-GalN/LPS-induced hepatic injury by decreasing hepatic toxicity, thereby mitigating the TLR 4-dependent cytokine release. The anti-inflammatory effect of AE could be contributing to that of AA and AE is better than AA.

Protective effect of ultrasonication-processed ginseng berry extract on the D-galactosamine/lipopolysaccharide-induced liver injury model in rats

  • Nam, Yoonjin;Bae, Jinhyung;Jeong, Ji Hoon;Ko, Sung Kwon;Sohn, Uy Dong
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.540-548
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    • 2018
  • Background: Acute hepatic failure is a life-threatening critical condition associated with rapid deterioration of liver function and liver transplantation. Several studies have shown that Panax ginseng Mayer has antidiabetic and hepatoprotective effects. However, the hepatoprotective effect of ginseng berry is still unveiled. In this study, we evaluated the hepatoprotective effects of ultrasonication-processed ginseng berry extract (UGBE) on acute hepatic failure model in rats. Methods: Ginseng berry extract (GBE) was ultrasonically processed. The GBE, silymarin, and UGBE were orally administered to male Sprague-Dawley rats for 4 wk. Twenty-four h after the last administration, rats were challenged with D-galactosamine (D-GalN)/lipopolysaccharide (LPS). Results: After ultrasonication, the component ratio of ginsenosides Rg2, Rg3, Rh1, Rh4, Rk1, Rk3, and F4 in GBE had been elevated. Administration of UGBE significantly increased the survival rate of D-GalN/LPS-challenged rats. Pretreatment with UGBE significantly decreased serum alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels in D-GalN/LPS-challenged rats in a dose-dependent manner. The levels of enzymatic markers for oxidative stress (superoxide dismutase, glutathione peroxidase, catalase, and glutathione) were increased by UGBE treatment in a dose-dependent manner. Tumor necrosis factor alphalevel, inducible nitric oxide synthase activities, and nitric oxide productions were reduced by UGBE treatment. In addition, hemeoxygenase-1 levels in liver were also significantly increased in the UGBE-treated group. The protein expression of toll-like receptor 4 was decreased by UGBE administration. Hematoxylin and eosin staining results also supported the results of this study showing normal appearance of liver histopathology in the UGBE-treated group. Conclusion: UGBE showed a great hepatoprotective effect on D-GalN/LPS-challenged rats via the toll-like receptor 4 signaling pathway.

Hepatoprotective Activities of Daidzin, Daidzein, Genistein and Puerarin in Primary Cultured Rat Hepatocytes (흰쥐의 일차배양 간세포에서 Daidzin, Daidzein, Genistein 및 Puerarin의 간 보호 활성 평가)

  • Park, Jin-Goo;Cheon, Ho-Joon;Kim, Yeong-Shik;Kang, Sam-Sik;Choi, Jae-Sue;Lee, Sun-Mee
    • YAKHAK HOEJI
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    • v.51 no.2
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    • pp.115-125
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    • 2007
  • The aim of this study was to investigate the protective activities of daidzin, daidzein, genistein or puerarin, active isoflavonoids of Puerariae Radix, on the hepatocyte injury induced by carbon tetrachloride (CCl$_4$, 10 mM), tert-butyl hydroperoxide (TBH, 0.5 mM) and D-galactosamine (GalN, 30 mM). Primary cultures of rat hepatocytes (18 hr cultured) were treated with CCl$_4$, TBH or GalN and various concentrations (0.1, 1, 10 and 100 ${\mu}$M) of daidzin, daidzein, genistein or puerarin. CCl$_4$ significantly increased the levels of lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The increase in LDH level was attenuated by daidzein, genistein and puerarin. Puerarin also inhibited the increase in AST level induced by CCl$_4$. The increases in LDH and ALT levels induced by TBH were significantly attenuated by daidzin and genistein treatments. GalN markedly increased the levels of LDH, ALT and AST These increases were significantly attenuated by daidzein. Daidzin also inhibited the increases in LDH and AST levels induced by GalN. The increases in LDH and ALT levels were attenuated by genistein and puerarin, respectively. These results suggest that daidzin and daidzein possess hepatoprotective activities.

Transcriptional activation of human GM3 synthase (hST3Gal V) gene by valproic acid in ARPE-19 human retinal pigment epithelial cells

  • Song, Na-Ree;Kim, Seok-Jo;Kwon, Haw-Young;Son, Sung-Wook;Kim, Kyoung-Sook;Ahn, Hee-Bae;Lee, Young-Choon
    • BMB Reports
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    • v.44 no.6
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    • pp.405-409
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    • 2011
  • The present study demonstrated that valproic acid (VPA) transcriptionally regulates human GM3 synthase (hST3Gal V), which catalyzes ganglioside GM3 biosynthesis in ARPE-19 human retinal pigment epithelial cells. For this, we characterized the promoter region of the hST3Gal V gene. Functional analysis of the 5'-flanking region of the hST3Gal V gene revealed that the -177 to -83 region functions as the VPA-inducible promoter and that the CREB/ATF binding site at -143 is crucial for VPA-induced expression of hST3Gal V in ARPE-19 cells. In addition, the transcriptional activity of hST3Gal V induced by VPA in ARPE-19 cells was inhibited by SP600125, a c-Jun N-terminal kinase (JNK) inhibitor. In summary, our results identified the core promoter region in the hST3Gal V promoter and for the first time demonstrated that ATF2 binding to the CREB/ATF binding site at -143 is essential for transcriptional activation of hST3Gal V in VPA-induced ARPE-19 cells.