猴头菇多糖对氧化应激的IPEC-J2细胞抗氧化能力及紧密连接蛋白ZO-1的影响

doi:10.11843/j.issn.0366-6964.2017.09.024

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (9): 1769-1776.doi: 10.11843/j.issn.0366-6964.2017.09.024

猴头菇多糖对氧化应激的IPEC-J2细胞抗氧化能力及紧密连接蛋白ZO-1的影响

陈新瑶¹, 张建龙², 董星¹, 陈景杰¹, 黄一帆^1*, 李健^1*

1. 福建农林大学福建省兽医中药与动物保健重点实验室, 福州 350002;
2. 中西兽医结合与动物保健福建省高等学校重点实验室, 福州 350002

收稿日期:2017-03-20 出版日期:2017-09-23 发布日期:2017-09-23
通讯作者: 黄一帆,E-mail:zjhyfang@163.com;李健,E-mail:lijian@fafu.edu.cn
作者简介:陈新瑶(1991-),男,硕士生,主要从事兽医中药药理学研究
基金资助:
国家自然科学基金（31502110）

Effects of Hericium Erinaceus Polysaccharide on Antioxidant Ability and ZO-1 Expression in IPEC-J2 Cells under Oxidative Stress

CHEN Xin-yao¹, ZHANG Jian-long², DONG Xing¹, CHEN Jing-jie¹, HUANG Yi-fan^1*, LI Jian^1*

1. Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health, Fujian Agriculture And Forestry University, Fuzhou 350002, China;
2. University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province, Fuzhou 350002, China

Received:2017-03-20 Online:2017-09-23 Published:2017-09-23

摘要/Abstract

摘要：

旨在探究在H₂O₂诱导的氧化应激损伤状态下，猴头菇多糖（HEPs）对IPEC-J2细胞的保护作用。将IPEC-J2细胞分为5组：空白组、模型组、HEPs低剂量组、HEPs中剂量组、HEPs高剂量组。采用MTT法分别选取试验用的H₂O₂和HEPs的最佳剂量；利用DCFH-DA染色观察并测定细胞内活性氧簇（ROS）的含量；通过比色法检测培养细胞上清液中丙二醛（MDA）的含量，细胞中超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活力以及乳酸脱氢酶（LDH）的释放率；应用实时荧光定量PCR（qRT-PCR）检测紧密连接蛋白ZO-1基因的转录水平。结果显示，（1）构建IPEC-J2细胞氧化应激模型的H₂O₂浓度为0.4 mmol·L^-1。（2） H₂O₂能够极显著地增加细胞内的ROS含量，诱导细胞凋亡，同时极显著地降低了SOD和CAT的活力，极显著地升高MDA含量与LDH释放率。但是，与模型组相比，经过HEPs预处理后，细胞内ROS的含量极显著地降低（P<0.01）；同时极显著地提高了SOD和CAT的活力（P<0.01），极显著地降低了MDA含量与LDH释放率（P<0.01）。（3）实时荧光定量PCR结果显示，H₂O₂能够极显著地降低ZO-1基因的转录量（P<0.01），与模型组比较，HEPs能够极显著的提高ZO-1基因的转录量（P<0.01）；Western blot结果显示ZO-1蛋白的表达量与ZO-1基因类似。可见，猴头菇多糖对H₂O₂诱导氧化损伤状态下的IPEC-J2细胞具有保护作用。

Abstract:

The objective of this study was to evaluate the protective effect of Hericium Erinaceus polysaccharide (HEPs) on intestinal epithelial cell injury induced by H₂O₂. In this study, IPEC-J2 cells were divided into five groups:the control group, the model group, the HEPs low dose treated group, the HEPs medium dose treated group, the HEPs high dose treated group. MTT method was used to select the optimal dose of H₂O₂ and HEPs, respectively; intracellular ROS was observed and determined by DCFH-DA staining; the content of malondialdehyde (MDA) in the supernatant of cultured cells and the activities of superoxide dismutase (SOD), catalase (CAT) and the release rate of lactate dehydrogenase (LDH) in cells were detected by colorimetric method; the transcription level of ZO-1 gene was detected by quantitative real-time PCR (qRT-PCR). The results showed that:(1) The 0.4 mmol·L^-1 was the most suitable H₂O₂ concentration for oxidative stress modeling. (2) The data indicated that H₂O₂ increased the intracellular ROS and induced cell apoptosis significantly, and decreased the vitalities of SOD and CAT significantly; H₂O₂ significantly increased the content of MDA and the release rate of LDH. But, compared with the model group, after HEPs pretreatment, the content of ROS in cells decreased significantly (P<0.01); at the same time, the activities of SOD and CAT were improved significantly (P<0.01), MDA content and LDH release rate were significantly decreased. (3) The results of quantitative real-time PCR displayed that H₂O₂ could significantly reduce the transcription of ZO-1 gene (P<0.01). Compared with H₂O₂ group, HEPs could significantly improve the transcription of ZO-1 genes after HEPs pretreatment (P<0.01). The results of Western blot showed that the expression of ZO-1 protein was similar to that of ZO-1 gene. Results showed that HEPs had protective effect on IPEC-J2 cells in H₂O₂ induced oxidative damage state.

中图分类号:

S856.4

陈新瑶, 张建龙, 董星, 陈景杰, 黄一帆, 李健. 猴头菇多糖对氧化应激的IPEC-J2细胞抗氧化能力及紧密连接蛋白ZO-1的影响[J]. 畜牧兽医学报, 2017, 48(9): 1769-1776.

CHEN Xin-yao, ZHANG Jian-long, DONG Xing, CHEN Jing-jie, HUANG Yi-fan, LI Jian. Effects of Hericium Erinaceus Polysaccharide on Antioxidant Ability and ZO-1 Expression in IPEC-J2 Cells under Oxidative Stress[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1769-1776.

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参考文献

[1] 吴永魁, 胡仲明. 动物应激医学及应激的分子调控机制[J]. 中国兽医学报, 2005, 25(5):557-560.
WU Y K, HU Z M. Animal stress medicine and the molecular mechanism of stress[J]. Chinese Journal of Veterinary Science, 2005, 25(5):557-560. (in Chinese)
[2] 顾立伟, 刘杨. 关于中国养猪业的思考[J]. 中国畜牧杂志, 2013, 49(2):50-54.
GU L W, LIU Y. Reflections on China's pig industry[J]. Chinese Journal of Animal Science, 2013, 49(2):50-54. (in Chinese)
[3] 刘亚华, 张杰, 代正, 等. 山莨菪碱对心搏骤停猪氧化应激致心肌线粒体损伤的保护作用[J]. 中华危重病急救医学, 2013, 25(5):290-293.
LIU Y H, ZHANG J, DAI Z, et al. Protection of anisodamine on the mitochondrial injury induced by oxidative stress in swine with cardiac arrest[J]. Chinese Critical Care Medicine, 2013, 25(5):290-293. (in Chinese)
[4] HE R R, LI Y, LI X D, et al. A new oxidative stress model, 2, 2-azobis (2-amidinopropane) dihydrochloride induces cardiovascular damages in chicken embryo[J]. PLoS One, 2013, 8(3):e57732.
[5] PIEKARSKI A L, KONG B W, LASSITER K, et al. Cell bioenergetics in Leghorn male hepatoma cells and immortalized chicken liver cells in response to 4-hydroxy 2-nonenal-induced oxidative stress[J]. Poult Sci, 2014, 93(11):2870-2877.
[6] WANG W W, WU Z L, LIN G, et al. Glycine stimulates protein synthesis and inhibits oxidative stress in pig small intestinal epithelial cells[J]. J Nutr, 2014, 144(10):1540-1548.
[7] WANG Y, TONG J, CHANG B, et al. Effects of alcohol on intestinal epithelial barrier permeability and expression of tight junction-associated proteins[J]. Mol Med Rep, 2014, 9(6):2352-2356.
[8] AL-SADI R, YE D M, BOIVIN M, et al. Interleukin-6 modulation of intestinal epithelial tight junction permeability is mediated by JNK pathway activation of claudin-2 gene[J]. PLoS One, 2014, 9(3):e85345.
[9] 高红梅. ETEC对断奶腹泻仔猪胃肠道紧密连接蛋白ZO-1的影响[D]. 武汉:华中农业大学, 2010.
GAO H M. Effects of ETEC with gastrointestinal tight junction protein ZO-1 on early weanling and diarrhea pigs[D]. Wuhan:Huazhong Agricultural University, 2010. (in Chinese)
[10] ZHU Y, CHEN Y, LI Q, et al. Preparation, characterization, and anti-Helicobacter pylori activity of Bi³⁺-Hericium erinaceus polysaccharide complex[J]. Carbohydr Polym, 2014, 110:231-237.
[11] ZAN X Y, CUI F J, LI Y H, et al. Hericium erinaceus polysaccharide-protein HEG-5 inhibits SGC-7901 cell growth via cell cycle arrest and apoptosis[J]. Int J Biol Macromol, 2015, 76:242-253.
[12] 邵梦茹. 猴头菇多糖对胃肠黏膜保护作用的实验研究[D]. 广州:广州中医药大学, 2014:3-4.
SHAO M R. Protective role of Hericium erinaceus polysaeeharides on gastrointestinal mucosal barrier function[D]. Guangzhou:Guangzhou University of Chinese Medicine, 2014:3-4. (in Chinese)
[13] 黄丽娜, 姜慧慧, 王洁, 等. 猴头菇多糖对呼肠孤病毒感染番鸭肠道黏膜免疫及抗氧化能力的影响[J]. 畜牧兽医学报, 2015, 46(9):1671-1677.
HUANG L N, JIANG H H, WANG J, et al. Effects of Hericium erinaceus polysaccharide on Intestinal mucosal immune and antioxidant capacity of Muscovy ducks infected with reovirus[J]. Acta Veterinaria et Zootechnica Sinica, 2015, 46(9):1671-1677. (in Chinese)
[14] 孙雪宁, 陈景杰, 黄诗杭, 等. 猴头菇多糖对MDRV感染雏番鸭肠道修复作用与MAdCAM-1分泌量关系的研究[J]. 中国家禽, 2016, 38(9):24-28.
SUN X N, CHEN J J, HUANG S H, et al. Relationship between intestinal tract restoration by Hericium erinaceus polysaccharide and MAdCAM-1 secretion in MDRV-infected Muscovy ducklings[J]. China Poultry, 2016, 38(9):24-28. (in Chinese)
[15] 陈新瑶, 董星, 陈景杰, 等. 响应曲面法优化猴头菇粗多糖的提取工艺[J]. 中国农业科技导报, 2017, 19(3):131-136.
CHEN X Y, DONG X, CHEN J J, et al. Optimization of extraction process for crude polysaccharide from Hericium erinaceus by response surface methodology[J]. Journal of Agricultural Science and Technology, 2017, 19(3):131-136. (in Chinese)
[16] ZHU L H, ZHAO K L, CHEN X L, et al. Impact of weaning and an antioxidant blend on intestinal barrier function and antioxidant status in pigs[J]. J Anim Sci, 2012, 90(8):2581-2589.
[17] PASZTI-GERE E, CSIBRIK-NEMETH E, SZEKER K, et al. Acute oxidative stress affects IL-8 and TNF-α expression in IPEC-J2 porcine epithelial cells[J]. Inflammation, 2012, 35(3):994-1004.
[18] ZHAO X M, XIANG H L, BAI X Y, et al. Porcine parvovirus infection activates mitochondria-mediated apoptotic signaling pathway by inducing ROS accumulation[J]. Virol J, 2016, 13:26.
[19] ALFONSO-PRIETO M, BIARNÉS X, VIDOSSICH P, et al. The molecular mechanism of the catalase reaction[J]. J Am Chem Soc, 2009, 131(33):11751-11761.
[20] KIN H, ZHAO Z Q, SUN H Y, et al. Postconditioning attenuates myocardial ischemia-reperfusion injury by inhibiting events in the early minutes of reperfusion[J]. Cardiovasc Res, 2004, 62(1):74-85.
[21] ANDERSON J M, VAN ITALLIE C M, FANNING A S. Setting up a selective barrier at the apical junction complex[J]. Curr Opin Cell Biol, 2004, 16(2):140-145.
[22] MATTER K, BALDA M S. Functional analysis of tight junctions[J]. Methods, 2003, 30(3):228-234.
[23] STEVENSON B R, SILICIANO J D, MOOSEKER M S, et al. Identification of ZO-1:A high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia[J]. J Cell Biol, 1986, 103(3):755-766.
[24] NASU Y, IDO A, TANOUE S, et al. Hepatocyte growth factor stimulates the migration of gastric epithelial cells by altering the subcellular localization of the tight junction protein ZO-1[J]. J Gastroenterol, 2013, 48(2):193-202.

猴头菇多糖对氧化应激的IPEC-J2细胞抗氧化能力及紧密连接蛋白ZO-1的影响

Effects of Hericium Erinaceus Polysaccharide on Antioxidant Ability and ZO-1 Expression in IPEC-J2 Cells under Oxidative Stress

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