L-Arginine Affects Immune Stressed Parameters and mRNA Expression of &beta;-Defensins Genes in Porcine Fetal Skin Fibroblasts

LUO Xue; ZHANG Chunyong; AN Qingcong; LI Meiquan; CHEN Kelin; GUO Rongfu

doi:10.3969/j.issn.1006-267x.2013.05.019

2013 , Vol. 25 >Issue 5: 1037 - 1044

DOI: https://doi.org/10.3969/j.issn.1006-267x.2013.05.019

Molecular Nutrition

L-Arginine Affects Immune Stressed Parameters and mRNA Expression of β-Defensins Genes in Porcine Fetal Skin Fibroblasts

LUO Xue ,
ZHANG Chunyong ,
AN Qingcong ,
LI Meiquan ,
CHEN Kelin ,
GUO Rongfu

Expand

Key Laboratory of Animal Nutrition and Feed Science of Yunnan Province, Yunnan Agricultural University, Kunming 650201, China

Received date: 2012-11-06

Online published: 2013-04-12

Fold

Abstract

This experiment was conducted to investigate the effects of L-arginine (L-Arg) on the immune stress parameters and mRNA expressions of β-defensins genes in porcine fetal skin fibroblasts (PFSF). The cultured 4 to 5 generations of PFSF were used, and immune stress model was induced by lipopolysaccharides (LPS). Adding different concentrations (0, 35, 70, 140, 280 and 560 μg/mL) of L-arginine in DMEM/F12 culture medium. After being cultured for 24 hours, the activities of lactate dehydrogenase (LDH), nitric oxide synthase (NOS) and lysozyme (LSZ) and nitric oxide (NO) content in culture medium were determined, and the mRNA expressions of β-defensin-1 (pBD-1), β-defensin-2 (pBD-2) and β-defensin-3 (pBD-3) genes were detected by the method of real-time qPCR. The results showed as follows: 1) L-arginine could reduce LDH activity in culture medium, and that in normal cells and stressed cells reached the lowest when L-arginine concentrations were 70 and 140 μg/mL, respectively; 2) L-arginine could significantly increase NO content in culture medium (P<0.01), and that in normal cells and stressed cells both reached the highest when L-arginine concentration was 140 μg/mL; 3) L-arginine could significantly increase NOS activity in normal cells (P<0.05 or P<0.01), but had no significant effects on stressed cells (P>0.05), when L-arginine concentrations were 35 and 70 μg/mL, NOS activity in stressed cells was significantly higher than that in normal cells; 4) the supplementation of 560 μg/mL L-arginine can significantly increase LSZ activity in culture medium (P>0.01); 5) the supplementation of 70 μg/mL L-arginine significantly increased the mRNA expressions of pBD-1 and pBD-2 genes, and the supplementation of 140 μg/mL L-arginine significantly increased the mRNA expression of pBD-3 gene. The results indicate that the proper concentration of L-arginine can decrease LDH activity, but increase NO content and the acitivities of NOS and LSZ in PFSF, and up-regulate the mRNA expressions of β-defensins genes; the optimal concentration of L-arginine is 70 to 140 μg/mL in the present study.

Key words： L-arginine; lipopolysaccharide; immnue stress; β-defensins; gene expression

Cite this article

LUO Xue , ZHANG Chunyong , AN Qingcong , LI Meiquan , CHEN Kelin , GUO Rongfu . L-Arginine Affects Immune Stressed Parameters and mRNA Expression of β-Defensins Genes in Porcine Fetal Skin Fibroblasts[J]. Chinese Journal of Animal Nutrition, 2013 , 25(5) : 1037 -1044 . DOI: 10.3969/j.issn.1006-267x.2013.05.019

References

[1] YANG D A,BIRAGYN L,KWAK W,et al.Mammalian defensins in immunity:more than just microbicidal[J].Trends in Immunology,2002,23(16):291-296.

[2] AONO S,LI C,ZHANG G,et al.Molecular and functional characterization of bovine beta-defensin-1[J].Veterinary Immunology and Immunopathology,2006,113(1/2):181-190.

[3] FEHLBAUM P,RAO M,ZASLOFF M,et al.An essential amino acid induces epithelial beta-defensin expression[J].Proceedings of the National Academy of Sciences of the United States of America,2000,97(23):12723-12728.

[4] 孟国权,刘玉兰,车政权,等.L-精氨酸对脂多糖诱导的断奶仔猪肠道黏膜屏障损伤的缓解作用[J].动物营养学报,2010,22(3):647-652.

[5] SHERMAN H,CHAPNIK N,FROY O.Albumin and amino acids upregulate the expression of human beta-defensin 1[J].Molecular Immunology,2006,43:1617-1623.

[6] PFAFFL M W.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Research,2001,29:2002-2007.

[7] HIBBS J B,TAINTOR R R,VAVRIN Z.Mcrophage cytotoxicity:role for L-arginine deminase and imino nitrogen oxidation to nitrite[J].Science,1987,235:473-476.

[8] 郑萍,田刚,毛湘冰,等.一氧化氮及其对营养物质代谢的调节[J].动物营养学报,2011,23(6):893-900.

[9] ALDERTON W K,COOPER C E,KNOWLES R G.Nitric oxide synthases:structure,fuction and inhibition[J].The Journal of Biochemistry,2001,357:593-615.

[10] LASKIN D L,DEL VALLE M R,HECK D E,et al.Hepatic nitric oxide production foloowing acute endotoxemia in rats is mediated by increased inducible nitric oxide synthase gene expression[J].Heptology,1995,22:223-224.

[11] MORRIS S M,Jr.Enzymes of arginine metabolism[J].The Journal of Nutrtion,2004,134:2743S-2747S.

[12] 骆雪,云南特有猪种β-defensin基因表达特点及外源活性物质的调节作用[D].硕士学位论文.昆明:云南农业大学,2012.

[13] JANEWAY C J,MEDZHITOV R.Innate immune recognition[J].Annual Review of Immunology,2002,20:197-216.

[14] DIAMOND G,RUSSELL J P,BEVINS C L.Inducible expression of an antibiotic peptide gene in lipopolysaccharide-challenged tracheal epitheial cell[J].Proceedings of the National Academy of Sciences of the United States of America,1996,93:5156-5160.

[15] 齐莎日娜.猪β-防御素基因表达特点及精氨酸的调节作用[D].博士学位论文.雅安:四川农业大学,2010.

[16] CAVERLY J M,DIAMOND G,GALLUP J M,et al.Coordinated expression of tracheal antimicrobial peptide and inflammatory-response elements in the lungs of neonatal calves with acute bacterial pneumonia[J].Infection and Immunity,2003,71:2950-2955.

[17] MEYERHOLZ D K,GALLUP J M,GRUBOR B M,et al.Developmental expression and distribution of sheep beta-defensin-2[J].Developmental and Comparative Immunology,2004,28(2):171-178.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References