不同壳聚糖浓度培养液对断奶仔猪外周血淋巴细胞中花生四烯酸代谢的影响

李俊良; 史彬林; 闫素梅; 金鹿; 徐元庆; 李倜宇; 郭祎玮; 郭晓宇

doi:10.3969/j.issn.1006-267x.2014.01.024

动物营养学报 >

2014 , Vol. 26 >Issue 1: 184 - 189

DOI: https://doi.org/10.3969/j.issn.1006-267x.2014.01.024

饲料营养 Feed Science and Technology

不同壳聚糖浓度培养液对断奶仔猪外周血淋巴细胞中花生四烯酸代谢的影响

李俊良 ,
史彬林 ,
闫素梅 ,
金鹿 ,
徐元庆 ,
李倜宇 ,
郭祎玮 ,
郭晓宇

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内蒙古农业大学动物科学学院, 呼和浩特 010018

收稿日期: 2013-08-14

网络出版日期: 2013-12-24

基金资助

国家自然科学基金项目（31060310）

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Effects of Different Chitosan Concentrations in Medium on Arachidonic Acid Metabolism in Peripheral Blood Lymphocytes of Weaner Piglets

LI Junliang ,
SHI Binlin ,
YAN Sumei ,
JIN Lu ,
XU Yuanqing ,
LI Tiyu ,
GUO Yiwei ,
GUO Xiaoyu

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College of Animal Science, Inner Mongolia Agricultural University, Huhhot 010018, China

Received date: 2013-08-14

Online published: 2013-12-24

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摘要

为了研究壳聚糖对断奶仔猪外周血淋巴细胞（PBLs）中花生四烯酸（AA）代谢的影响，试验分离提取断奶仔猪PBLs，在含有0、40、80、160和320 μg/mL壳聚糖的培养液中进行培养48 h后，测定培养液上清液中胞浆型磷脂酶A2（cPLA2）、环氧合酶-2（COX-2）和5-脂氧化酶（5-LOX）的活性及AA、前列腺素E2（PGE2）和白三烯B4（LTB4）的浓度；测定细胞中cPLA2、COX-2和5-LOX mRNA的相对表达量。结果表明：随着壳聚糖添加水平的增加，淋巴细胞培养液中的cPLA2、COX-2和5-LOX的活性及AA、PGE2和LTB4浓度呈显著的二次曲线变化（P<0.05）；淋巴细胞中的cPLA2、COX-2和5-LOX mRNA的相对表达量与壳聚糖添加水平也呈显著的二次剂量依赖关系（P<0.05）。上述所有指标的最大值出现在壳聚糖的添加水平为80~160 μg/mL时，而当壳聚糖的添加水平增加到320 μg/mL时其值有不同程度的降低。结果提示，在本试验中，适当水平的壳聚糖可以促进断奶仔猪PBLs内AA代谢网络途径中参与炎症或免疫调节的主要介质PGE2和LTB4的产生、关键控制酶cPLA2、COX-2和5-LOX的活性及其基因表达，这可能是其发挥免疫调节功能的原因之一。

关键词： 壳聚糖; 断奶仔猪; 外周血淋巴细胞; 花生四烯酸代谢; 免疫功能

本文引用格式

李俊良 , 史彬林 , 闫素梅 , 金鹿 , 徐元庆 , 李倜宇 , 郭祎玮 , 郭晓宇 . 不同壳聚糖浓度培养液对断奶仔猪外周血淋巴细胞中花生四烯酸代谢的影响[J]. 动物营养学报, 2014 , 26(1) : 184 -189 . DOI: 10.3969/j.issn.1006-267x.2014.01.024

Abstract

This experiment was conducted to investigate the effects of different levels of chitosan in medium on the metabolism of arachidonic acid in peripheral blood lymphocytes of weaner piglet. The peripheral blood lymphocytes from weaner piglets were cultured in medium supplemented with 0 (control), 40, 80, 160 and 320 μg chitosan/mL medium, respectively. At the end of 48 h incubation, supernatants were collected for analyzing the activities of cytosolic phospholipase A2 (cPLA2), cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) as well as the concentrations of arachidonic acid (AA), prostaglandin E2 (PGE2) and leukotriene B4 (LTB4). The cells were harvested and analyzed for the mRNA expressions of cPLA2, COX-2 and 5-LOX genes. Results showed that chitosan increased the activities of cPLA2, COX-2 and 5-LOX as well as the concentrations of AA, PGE2 and LTB4 in supernatants in a quadratic dose-dependent manner (P<0.05). With the chitosan supplemental level increasing, the mRNA expressions of cPLA2, COX-2 and 5-LOX genes in peripheral blood lymphocytes was enhanced in a quadratic manner (P<0.05). In addition, the supplementation of 80 to 160 μg/mL chitosan to the medium had higher values of all above indexes compared with those in other treatments. However, positive effects of chitosan tended to be suppressed when its supplemental level was increased to 320 μg/mL. It is concluded that appropriate level of chitosan can increase the concentrations of AA, PGE2 and LTB4, and improve the activities of cPLA2, COX-2 and 5-LOX as well as the mRNA expressions of cPLA2, COX-2 and 5-LOX genes in peripheral blood lymphocytes, which may be one of the reasons for chitosan to regulate the immune functions of weaner piglets.

Key words： chitosan; weaner piglet; peripheral blood lymphocytes; arachidonic acid metabolism; immune function

参考文献

[1] LI L L, YIN F G, ZHANG B, et al.Dietary supplementation with Atractylodes macrophala Koidz polysaccharides ameliorate metabolic status and improve immune function in early-weaned pigs[J].Livestock Science, 2011, 142(1/2/3):33-41.

[2] WANG X Q, FENG Y, SHU G, et al.Effect of dietary supplementation with hydrolyzed wheat gluten on growth performance, cell immunity and serum biochemical indices of weaned piglets(Sus scrofa)[J].Agricultural Sciences in China, 2011, 10(6):938-945.

[3] 韩杰, 边连全, 刘显军, 等.刺五加多糖对断奶仔猪血液免疫指标的影响[J].动物营养学报, 2012, 24(12):2444-2449.

[4] KONG X F, YIN Y L, HE Q H, et al.Dietary supplementation with Chinese herbal powder enhances ileal digestibilities and serum concentrations of amino acids in young pigs[J].Amino Acids, 2009, 37(4):573-582.

[5] WANG M Q, DU Y J, WANG C, et al.Effects of copper-loaded chitosan nanoparticles on intestinal microflora and morphology in weaned piglets[J].Biological Trace Element Research, 2012, 49(2):184-189.

[6] 张彩芬, 史彬林, 金晓, 等.壳聚糖对仔猪生长性能及血清中类胰岛素生长因子-Ⅰ和生长激素水平的影响[J].动物营养学报, 2008, 20(2):191-195.

[7] 刘莹, 陈政, 尚尔昌, 等.花生四烯酸代谢网络研究:从关键酶的单靶标抑制剂到多靶标抑制剂[J].药学学报, 2009, 44(3):231-241.

[8] 唐春华, 陈韬.花生四烯酸生物活性及其对机体的免疫作用[J].畜禽业, 2009(6):20-22.

[9] 彭碧文, 黄清玲, 林建银, 等.外源性花生四烯酸在弓形虫入侵巨噬细胞中的信号作用[J].中国寄生虫学与寄生虫病杂志, 2003, 21(5):296-299.

[10] 袁成凌, 姚建铭, 余增亮.花生四烯酸及其代谢物的生物学作用[J].中国药物化学杂志, 2000, 10(1):75-78.

[11] BIANEO I D, BALSINDE J, BELTRAMO D M, et al.Chitosan-induced phospholipase A2 activation and arachidonic acid mobilization in P388D1 macrophages[J].FEBS Letters, 2000, 466(2/3):292-294.

[12] USAMI Y, OKAMOTO Y, TAKAYAMA T, et al.Chitin and chitosan stimulate canine polymorphonuclear cells to release leukotriene B₄ and prostaglandin E₂[J].Journal of Biomedical Materials Research, 1998, 42(4):517-522.

[13] 李慧英, 史彬林, 闫素梅, 等.壳聚糖对肉仔鸡血清花生四烯酸含量、磷脂酶A2活性及小肠胞内磷脂酶A2 mRNA表达的影响[J].动物营养学报, 2009, 21(4):580-584.

[14] KENNETH J L, THOMAS D S.Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCt method[J].Methods, 2001, 25(4):402-408.

[15] 罗大力, 张幼怡, 韩启德.花生四烯酸的生物活性及其钙信号转导作用[J].生理科学进展, 2002, 33(3):251-254.

[16] 杨磊, 孟庆雄.磷脂酶A2的研究进展[J].食品与药品, 2007, 9(7):52-54.

[17] 李慧英.壳聚糖对肉仔鸡免疫功能的影响及其分子机理的研究[D].博士学位论文.呼和浩特:内蒙古农业大学, 2009:54-63.

[18] CHOA Y S, LEEA S H, KIMA S K.Aminoethyl-chitosan inhibits LPS-induced inflammatory mediators, iNOS and COX-2 expression in RAW264.7 mouse macrophages[J].Process Biochemistry, 2011, 46(2):465-470.

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