发酵麸皮多糖对大鼠空肠组织抗氧化能力、形态结构和紧密连接蛋白表达的影响
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摘要
目的:研究发酵麸皮多糖对断奶大鼠空肠组织抗氧化能力、形态结构和紧密连接蛋白表达的影响。方法:将40只雄性断奶Wistar大鼠随机分为5组,包括对照组、VC组及发酵麸皮多糖低、中、高剂量组(每天分别灌服生理盐水、100 mg/kg mb VC及20、40、80 mg/kg mb发酵麸皮多糖,灌胃剂量均为0.4 mL),实验期为21 d。检测大鼠空肠组织中总抗氧化能力(total antioxitant capacity,T-AOC),过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活力以及谷胱甘肽(glutathione,GSH)、8-羟基脱氧鸟苷(8-hydroxy deoxyguanosine,8-OHd G)含量;采用苏木素-伊红染色法分析大鼠空肠测定绒毛高度和隐窝深度,计算绒毛高度与隐窝深度的比值;并采用实时荧光定量聚合酶链反应法测定空肠Claudin-1、Occludin和ZO-1 3种紧密连接蛋白的m RNA表达量。结果:与对照组相比,灌胃40 mg/kg mb发酵麸皮多糖能显著提高断奶大鼠空肠组织中T-AOC、CAT、SOD和GSH-Px活力以及GSH含量(P<0.05),显著降低8-OHdG含量(P<0.05);提高空肠绒毛高度和绒毛高度/隐窝深度,降低隐窝深度;提高Claudin-1、Occludin和ZO-1 3种紧密连接蛋白的mRNA表达量。结论:发酵麸皮多糖可以改善断奶大鼠空肠抗氧化能力和形态结构,提高紧密连接蛋白的表达,起到保护肠道健康的作用。
This study was conducted to investigate the effects of oral supplementation with fermented wheat bran polysaccharides(FWBPs) on antioxidant capacity,morphological structure and tight junction protein expression in the jejunum of weaned rat pups.Forty healthy weaned Wistar rats with similar body mass were randomly assigned to five groups,including control(saline),VC(positive control,100 mg/kg mb),and low-,medium-and high-dose FWBPs groups(20、40、80 mg/kg mb in a dosage of 0.4 mL).The experimental duration was 21 days.The total antioxidative capacity(T-AOC),catalase(CAT),superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) activities,and glutathione(GSH) and 8-hydroxydeoxyguanosine(8-OHdG) contents in jejunum were measured.Histopathological examination with hematoxylin and eosin(HE) staining was used to analyze villus height and crypt depth,and the ratio of villus height to crypt depth was calculated.The mRNA expression of the tight junction proteins Claudin-1,Occludin and zonula occludens-1 were determined using quantitative real-time polymerase chain reaction(qPCR).The results showed that oral administration of FWBPs at 40 mg/kg mb significantly improved T-AOC,CAT,SOD and GSH-Px activities and GSH content in jejunum(P<0.05),and significantly decreased the content of 8-OHdG(P<0.05).FWBPs increased the villus height and decreased crypt depth,thus resulting in increased villus height/crypt depth ratio.The mRNA expression of the tight junction proteins in the jejunum was significantly up-regulated in the FWBPs groups.In summary,fermented wheat bran polysaccharides can improve the antioxidant capacity and jejunal morphology and promote the expression of tight junction proteins,thereby preserving intestinal health in weaned rats.
This study was conducted to investigate the effects of oral supplementation with fermented wheat bran polysaccharides(FWBPs) on antioxidant capacity,morphological structure and tight junction protein expression in the jejunum of weaned rat pups.Forty healthy weaned Wistar rats with similar body mass were randomly assigned to five groups,including control(saline),VC(positive control,100 mg/kg mb),and low-,medium-and high-dose FWBPs groups(20、40、80 mg/kg mb in a dosage of 0.4 mL).The experimental duration was 21 days.The total antioxidative capacity(T-AOC),catalase(CAT),superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) activities,and glutathione(GSH) and 8-hydroxydeoxyguanosine(8-OHdG) contents in jejunum were measured.Histopathological examination with hematoxylin and eosin(HE) staining was used to analyze villus height and crypt depth,and the ratio of villus height to crypt depth was calculated.The mRNA expression of the tight junction proteins Claudin-1,Occludin and zonula occludens-1 were determined using quantitative real-time polymerase chain reaction(qPCR).The results showed that oral administration of FWBPs at 40 mg/kg mb significantly improved T-AOC,CAT,SOD and GSH-Px activities and GSH content in jejunum(P<0.05),and significantly decreased the content of 8-OHdG(P<0.05).FWBPs increased the villus height and decreased crypt depth,thus resulting in increased villus height/crypt depth ratio.The mRNA expression of the tight junction proteins in the jejunum was significantly up-regulated in the FWBPs groups.In summary,fermented wheat bran polysaccharides can improve the antioxidant capacity and jejunal morphology and promote the expression of tight junction proteins,thereby preserving intestinal health in weaned rats.
引文
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[9]周素梅,曹丽,王强,等.小麦麸皮中碱法与酶法提取阿拉伯木聚糖的结构与免疫活性比较研究[C]//中国食品科学技术学会第六届年会暨第五届东西方食品业高层论坛论文摘要集.北京:中国食品科学技术学会,2010:237.
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[14]刘丽娅,赵梦丽,钟葵,等.小麦麸皮阿拉伯木聚糖体外益生活性研究[J].中国粮油学报,2016,31(10):1-5;30.DOI:10.3969/j.issn.1003-0174.2016.10.001.
[15]陈群,乐国伟,施用晖,等.氧自由基对动物消化道损伤及干预研究进展[J].中国畜牧兽医,2006,33(11):106-108.DOI:10.3969/j.issn.1671-7236.2006.11.039.
[16]安晓萍,王园,齐景伟,等.基于响应面法分析菌比和辅料对发酵麸皮多糖含量的影响[J].食品工业科技,2018,39(3):122-126.DOI:10.13386/j.issn1002-0306.2018.03.025.
[17]王园,史俊祥,段元霄,等.麸皮多糖微生物发酵工艺优化及其抗炎活性[J].食品科学,2018,39(14):192-198.DOI:10.7506/spkx1002-6630-201814029.
[18]安晓萍,王园,史俊祥,等.一种发酵麸皮多糖的提取及其对大鼠抗氧化功能的影响[J].食品工业科技,2018,39(16):281-285.DOI:10.13386/j.issn1002-0306.2018.16.050.
[19]廖波.25-OH-D3对免疫应激断奶仔猪的生产性能,肠道免疫功能和机体免疫应答的影响[D].雅安:四川农业大学,2011:28-29.
[20]王秋林,王浩毅,王树人.氧化应激状态的评价[J].中国病理生理杂志,2005,21(10):2069-2074.DOI:10.3321/j.issn:1000-4718.2005.10.049.
[21]魏涛,高玉鹏,杜忍让,等.复合酶制剂对蛋鸡血清抗氧化酶系与肠道形态学影响[J].饲料与畜牧,2011(1):13-16.
[22]段元霄,王园,史俊祥,等.麦麸阿魏酰低聚糖对敌草快致氧化应激大鼠血浆和组织抗氧化能力的影响[J].动物营养学报,2017,29(12):4648-4658.DOI:10.3969/j.issn.1006-267x.2017.12.048.
[23]毛湘冰,陈代文,胡海燕,等.饲粮中添加香菇多糖对断奶大鼠生长性能、抗氧化能力和肠道健康的影响[J].动物营养学报,2017,29(3):859-865.DOI:10.3969/j.issn.1006-267x.2017.03.016.
[24]CHEN H,MAO X B,HE J,et al.Dietary fibre affects intestinal mucosal barrier function and regulates intestinal bacteria in weaning piglets[J].British Journal of Nutrition,2013,110(10):1837-1848.DOI:10.1017/S0007114513001293.
[25]CASPARY W F.Physiology and pathophysiology of intestinal absorption[J].American Journal of Clinical Nutrition,1992,55(1):2995-3085.DOI:10.1093/ajcn/55.1.299s.
[26]韩正康.家畜营养生理学[M].北京:农业出版社,1991:16-17.
[27]顾宪红,张宏福,佘锐萍,等.断奶日龄对消化器官形态及小肠组织化学的影响[J].家畜生态学报,2003,24(1):24-30.DOI:10.3969/j.issn.1673-1182.2003.01.008.
[28]江昌盛,周彤,张倩,等.香菇多糖对大肠杆菌攻毒大鼠空肠形态结构、上皮细胞数量和紧密连接蛋白表达的影响[J].动物营养学报,2017,29(10):3592-3597.DOI:10.3969/j.issn.1006-267x.2017.10.020.
[29]FURUSE M.Molecular basis of the core structure of tight junctions[J].Cold Spring Harbor Perspectives in Biology,2010,2(1):1-16.DOI:10.1101/cshperspect.a002907.
[30]SHEN L,WEBER C R,RALEIGH D R,et al.Tight junction pore and leak pathways:a dynamic duo[J].Annual Review of Physiology,2011,73:283-309.DOI:10.1146/annurev-physiol-012110-142150.
[2]张梅红,钟葵,刘丽娅,等.小麦麸皮营养与质量安全品质分析[J].麦类作物学报,2012,32(6):1090-1095.DOI:10.7606/j.issn.1009-1041.2012.06.014.
[3]蔺艳君,刘丽娅,钟葵,等.不同来源小麦麸皮营养成分及酚类物质含量的比较[J].现代食品科技,2014,30(12):194-200.DOI:10.13982/j.mfst.1673-9078.2014.12.033.
[4]MAES C,DELEOUR A.Structoral characterization of water-extractable and water-unextractable arabinoxylans in wheat bran[J].Journal Cereal Science,2002,35:315-326.DOI:10.1016/j.jcs.2010.09.002.
[5]王吉中,冯昕,冯永强,等.麦麸多糖提取工艺及其抑菌活性的研究[J].食品工业,2011,32(11):1-4.
[6]王忠合,钟丽娴.麦麸活性多糖的提取、组成及其抗氧化性研究[J].食品工业科技,2009,30(7):115-119.DOI:10.13386/j.issn1002-0306.2009.07.095.
[7]HIGUCHI M,OSHIDA J,ORINO K,et al.Wheat bran protects Fischer-344 rats from diquat-induced oxidative stress by activating antioxidant system:selenium as an antioxidant[J].Bioscience Biotechnology and Biochemistry,2011,75(3):496-499.DOI:10.1271/bbb.100719.
[8]YUAN X P,WANG J,YAO H Y,et al.Antioxidant activity of feruloylated oligosaccharides from wheat bran[J].Food Chemistry,2005,90(4):759-764.DOI:10.1016/j.foodchem.2004.05.025.
[9]周素梅,曹丽,王强,等.小麦麸皮中碱法与酶法提取阿拉伯木聚糖的结构与免疫活性比较研究[C]//中国食品科学技术学会第六届年会暨第五届东西方食品业高层论坛论文摘要集.北京:中国食品科学技术学会,2010:237.
[10]CAO L,LIU X Z,QIAN T X,et al.Antitumor and immunomodulatory activity of arabinoxylans:a major constituent of wheat bran[J].International Journal of Biological Macromolecules,2011,48(1):160-164.DOI:10.1016/j.ijbiomac.2010.10.014.
[11]ZHOU S M,LIU X Z,GUO Y,et al.Comparison of the immunological activities of arabinoxylans from wheat bran with alkali and xylanase-aided extraction[J].Carbohydrate Polymers,2010,81(4):784-789.DOI:10.1016/j.carbpol.2010.03.040.
[12]YANG J,MALDONADOGOMEZ M X,HUTKINS R W,et al.Production and in vitro fermentation of soluble,non-digestible,feruloylated oligo-and polysaccharides from maize and wheat brans[J].Journal of Agricultural and Food Chemistry,2014,62(1):159-166.DOI:10.1021/jf404305y.
[13]周迎春.小麦麸皮低聚糖生产工艺优化及其功能特性研究[D].郑州:河南工业大学,2014:26-38.
[14]刘丽娅,赵梦丽,钟葵,等.小麦麸皮阿拉伯木聚糖体外益生活性研究[J].中国粮油学报,2016,31(10):1-5;30.DOI:10.3969/j.issn.1003-0174.2016.10.001.
[15]陈群,乐国伟,施用晖,等.氧自由基对动物消化道损伤及干预研究进展[J].中国畜牧兽医,2006,33(11):106-108.DOI:10.3969/j.issn.1671-7236.2006.11.039.
[16]安晓萍,王园,齐景伟,等.基于响应面法分析菌比和辅料对发酵麸皮多糖含量的影响[J].食品工业科技,2018,39(3):122-126.DOI:10.13386/j.issn1002-0306.2018.03.025.
[17]王园,史俊祥,段元霄,等.麸皮多糖微生物发酵工艺优化及其抗炎活性[J].食品科学,2018,39(14):192-198.DOI:10.7506/spkx1002-6630-201814029.
[18]安晓萍,王园,史俊祥,等.一种发酵麸皮多糖的提取及其对大鼠抗氧化功能的影响[J].食品工业科技,2018,39(16):281-285.DOI:10.13386/j.issn1002-0306.2018.16.050.
[19]廖波.25-OH-D3对免疫应激断奶仔猪的生产性能,肠道免疫功能和机体免疫应答的影响[D].雅安:四川农业大学,2011:28-29.
[20]王秋林,王浩毅,王树人.氧化应激状态的评价[J].中国病理生理杂志,2005,21(10):2069-2074.DOI:10.3321/j.issn:1000-4718.2005.10.049.
[21]魏涛,高玉鹏,杜忍让,等.复合酶制剂对蛋鸡血清抗氧化酶系与肠道形态学影响[J].饲料与畜牧,2011(1):13-16.
[22]段元霄,王园,史俊祥,等.麦麸阿魏酰低聚糖对敌草快致氧化应激大鼠血浆和组织抗氧化能力的影响[J].动物营养学报,2017,29(12):4648-4658.DOI:10.3969/j.issn.1006-267x.2017.12.048.
[23]毛湘冰,陈代文,胡海燕,等.饲粮中添加香菇多糖对断奶大鼠生长性能、抗氧化能力和肠道健康的影响[J].动物营养学报,2017,29(3):859-865.DOI:10.3969/j.issn.1006-267x.2017.03.016.
[24]CHEN H,MAO X B,HE J,et al.Dietary fibre affects intestinal mucosal barrier function and regulates intestinal bacteria in weaning piglets[J].British Journal of Nutrition,2013,110(10):1837-1848.DOI:10.1017/S0007114513001293.
[25]CASPARY W F.Physiology and pathophysiology of intestinal absorption[J].American Journal of Clinical Nutrition,1992,55(1):2995-3085.DOI:10.1093/ajcn/55.1.299s.
[26]韩正康.家畜营养生理学[M].北京:农业出版社,1991:16-17.
[27]顾宪红,张宏福,佘锐萍,等.断奶日龄对消化器官形态及小肠组织化学的影响[J].家畜生态学报,2003,24(1):24-30.DOI:10.3969/j.issn.1673-1182.2003.01.008.
[28]江昌盛,周彤,张倩,等.香菇多糖对大肠杆菌攻毒大鼠空肠形态结构、上皮细胞数量和紧密连接蛋白表达的影响[J].动物营养学报,2017,29(10):3592-3597.DOI:10.3969/j.issn.1006-267x.2017.10.020.
[29]FURUSE M.Molecular basis of the core structure of tight junctions[J].Cold Spring Harbor Perspectives in Biology,2010,2(1):1-16.DOI:10.1101/cshperspect.a002907.
[30]SHEN L,WEBER C R,RALEIGH D R,et al.Tight junction pore and leak pathways:a dynamic duo[J].Annual Review of Physiology,2011,73:283-309.DOI:10.1146/annurev-physiol-012110-142150.