硒对雏鸡空肠和回肠IL-1β、IL-6和TNF-α mRNA表达的影响
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摘要
通过复制缺硒雏鸡模型,对雏鸡空肠和回肠中主要细胞因子的变化进行不同时间点的检测,为硒调控肠黏膜免疫的机制研究奠定基础。选取120只1日龄的SPF雏鸡,随机分为对照组和试验组2组。对照组雏鸡饲喂全价饲料,试验组雏鸡饲喂缺硒饲料。分别在试验期7,14,21,28d对雏鸡进行剖杀并取其空肠与回肠,利用实时荧光定量PCR法检测IL-1β、IL-6和TNF-α的mRNA表达水平。结果显示,试验组与对照组相比,缺硒雏鸡空肠在试验期14,21d时,IL-1β、IL-6和TNF-α细胞因子的表达量均升高,且差异极显著(P<0.01);28d时,除IL-1β的表达量降低(P<0.05),其他细胞因子的相对表达量升高(P<0.01)。试验组与对照组相比,缺硒雏鸡回肠在试验期14,21d时,IL-1β、IL-6和TNF-α细胞因子表达量均升高(P<0.01);在28d时,IL-6和TNF-α的相对表达量升高(P<0.05)。结果表明,缺硒能够上调肠黏膜炎性细胞因子IL-1β、IL-6和TNF-αmRNA的相对表达量,这可能与硒缺乏导致雏鸡肠黏膜中氧自由基含量升高有关。
In this study,the selenium-deficient chicken model was duplicated to detect the changes of major cytokines in the jejunum and ileum of chickens at different time points,which laid the foundation for the study on the mechanism of selenium-mediated intestinal mucosal immunity.120one-day-old SPF chickens were selected and randomly divided into control group and experimental group.The control group chickens fed the full-fledged feed and the experimental group chickens fed the selenium-deficient feed.The chickens were killed on the 7th,14 th,21st,and 28 th day of the experiment,and the jejunum and ileum of the chickens were taken.The mRNA expression levels of IL-1β,IL-6and TNF-α were detected by real-time fluorescence quantitative PCR.The results showed that compared with the control group,the expression of IL-1β,IL-6and TNF-αcytokines in jejunum of selenium-deficient chickens increased significantly on the 14 th and 21 st day of the experimental period(P<0.01).On the 28 th day,the expression of IL-1βwas decreased(P<0.05),and the relative expression of other cytokines increased(P<0.01).Compared with the control group,the IL-1β,IL-6and TNF-αcytokines in the ileum of the selenium-deficient chicken were all increased at the 14 th and 21 st days of the experimental period(P<0.01).At 28 days,the relative expression of IL-6and TNF-αincreased(P<0.05).Therefore,selenium deficiency can upregulate the expression of inflammatory cytokines IL-1β,IL-6and TNF-α mRNA in intestinal mucosa,which may be related to the lack of selenium leading to the increase of oxygen free radicals in the intestinal mucosa.
In this study,the selenium-deficient chicken model was duplicated to detect the changes of major cytokines in the jejunum and ileum of chickens at different time points,which laid the foundation for the study on the mechanism of selenium-mediated intestinal mucosal immunity.120one-day-old SPF chickens were selected and randomly divided into control group and experimental group.The control group chickens fed the full-fledged feed and the experimental group chickens fed the selenium-deficient feed.The chickens were killed on the 7th,14 th,21st,and 28 th day of the experiment,and the jejunum and ileum of the chickens were taken.The mRNA expression levels of IL-1β,IL-6and TNF-α were detected by real-time fluorescence quantitative PCR.The results showed that compared with the control group,the expression of IL-1β,IL-6and TNF-αcytokines in jejunum of selenium-deficient chickens increased significantly on the 14 th and 21 st day of the experimental period(P<0.01).On the 28 th day,the expression of IL-1βwas decreased(P<0.05),and the relative expression of other cytokines increased(P<0.01).Compared with the control group,the IL-1β,IL-6and TNF-αcytokines in the ileum of the selenium-deficient chicken were all increased at the 14 th and 21 st days of the experimental period(P<0.01).At 28 days,the relative expression of IL-6and TNF-αincreased(P<0.05).Therefore,selenium deficiency can upregulate the expression of inflammatory cytokines IL-1β,IL-6and TNF-α mRNA in intestinal mucosa,which may be related to the lack of selenium leading to the increase of oxygen free radicals in the intestinal mucosa.
引文
[1]袁丽君,袁林喜,尹雪斌,等.硒的生理功能、摄入现状与对策研究进展[J].生物技术进展,2016,6(6):396-405.
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[12]FIDLER J W,NABER E C,LATSHAW J D.Effect of peroxide administration on selenium utilization,growth,deficiency symptoms,and glutathione peroxidase activity in chicks fed controlled selenium diets[J].Poult Sci,1980,59(1):141-148.
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[14]HEFNAWY A E G,TORTORA-PEREZ J L.The importance of selenium and the effects of its deficiency in animal health[J].Small Ruminant Res,2010,89(2):185-192.
[15]KANG J H,BAIK H W,CHUNG H K.Trace element deficiencies in long-term tube fed patients[J].eSPEN J,2014,9(2):e63-e68.
[16]王哲.动物营养代谢病[M].吉林:吉林科学技术出版社,1995.
[17]刘宗平.现代动物营养代谢病[M].北京:化学工业出版社,2003.
[18]HANGALAPURA B N,KAISER M G,VAN DERPOEL J J,et al.Cold stress equally enhances in vivo pro-inflammatory cytokine gene expression in chicken lines divergently selected for antibody responses[J].Dev Comp Immunol,2006,30(5):503-511.
[19]MEYER O.Role of TNF-alpha and cytokines in the physiopathology of rheumatoid arthritis.Therapeutic perspectives[J].Bull Acad Natl Med,2002,187(5):935-954.
[20]仝宗喜.雏鸡硒缺乏症分子机理的研究[D].黑龙江哈尔滨:东北农业大学,2004.
[21]宋代琴,王治伦,陈静宏,等.低硒条件下T-2毒素对大鼠血液中细胞因子IL-6,IL-1β和TNF-α水平的影响[J].中国地方病防治杂志,2009,24(5):327-330.
[22]由路.硒蛋白T与鸡缺硒性免疫功能障碍的关系[D].黑龙江哈尔滨:东北农业大学,2014.
[23]ZHANG W,ZHANG R,WANG T,et al.Selenium inhibits LPS-induced pro-inflammatory gene expression by modulating MAPK and NF-κB signaling pathways in mouse mammary epithelial cells in primary culture[J].Inflammation,2014,37(2):478-485.
[2]陈彦霖,范艳丽.硒的生物学功能及富硒食品的研究进展[J].河北农业科学,2008,12(9):99-100.
[3]VALDIGLESIAS V.In vitro evaluation of selenium genotoxic,cytotoxic,and protective effects:a review[J].Arch Toxicol,2010,84(5):337-351.
[4]李晓丽,何万领,文凤云,等.酵母硒对蛋鸡血清酶活性及抗氧化能力的影响[J].中国兽医学报,2017,37(2):318-321.
[5]NAVARRO-ALARCON M,LPEZ-MARTNEZ MC.Essentiality of selenium in the human body:relationship with different diseases[J].Sci Total Environ,2000,249(1):347-371.
[6]PAPPAS A C,ZOIDIS E,SURAI P F,et al.Selenoproteins and maternal nutrition[J].Comp Biochem Physiol B Biochem Mol Biol,2008,151(4):361-372.
[7]SURAI P F.Effect of selenium and vitamin E content of the maternal diet on the antioxidant system of the yolk and the developing chick[J].Br Poult Sci,2000,41(2):235-243.
[8]荔霞,刘永明,齐志明,等.硒对小鼠血清中细胞因子水平的调控作用研究[J].中国畜牧兽医,2007,34(6):98-100.
[9]OZBAL S,ERBIL G,KOCDOR H,et al.The effects of selenium against cerebral ischemia-reperfusion injury in rats[J].Neurosci Lett,2008,438(3):265-269.
[10]尹希海,于巧玲,李灵.缺硒地区仔猪腹泻的防治[J].中国兽医杂志,2000,26(12):46-47.
[11]HANGALAPURA B N,KAISER M G,VAN DERPOEL J J,et al.Cold stress equally enhances in vivo pro-inflammatory cytokine gene expression in chicken lines divergently selected for antibody responses[J].Dev Comp Immunol,2006,30(5):503-511.
[12]FIDLER J W,NABER E C,LATSHAW J D.Effect of peroxide administration on selenium utilization,growth,deficiency symptoms,and glutathione peroxidase activity in chicks fed controlled selenium diets[J].Poult Sci,1980,59(1):141-148.
[13]LEYCK S.Pamham-Acute anti inflammatory and gastric effects of the selen organic compound ebselen[J].Agents Actions,1990,30(3/4):426-431.
[14]HEFNAWY A E G,TORTORA-PEREZ J L.The importance of selenium and the effects of its deficiency in animal health[J].Small Ruminant Res,2010,89(2):185-192.
[15]KANG J H,BAIK H W,CHUNG H K.Trace element deficiencies in long-term tube fed patients[J].eSPEN J,2014,9(2):e63-e68.
[16]王哲.动物营养代谢病[M].吉林:吉林科学技术出版社,1995.
[17]刘宗平.现代动物营养代谢病[M].北京:化学工业出版社,2003.
[18]HANGALAPURA B N,KAISER M G,VAN DERPOEL J J,et al.Cold stress equally enhances in vivo pro-inflammatory cytokine gene expression in chicken lines divergently selected for antibody responses[J].Dev Comp Immunol,2006,30(5):503-511.
[19]MEYER O.Role of TNF-alpha and cytokines in the physiopathology of rheumatoid arthritis.Therapeutic perspectives[J].Bull Acad Natl Med,2002,187(5):935-954.
[20]仝宗喜.雏鸡硒缺乏症分子机理的研究[D].黑龙江哈尔滨:东北农业大学,2004.
[21]宋代琴,王治伦,陈静宏,等.低硒条件下T-2毒素对大鼠血液中细胞因子IL-6,IL-1β和TNF-α水平的影响[J].中国地方病防治杂志,2009,24(5):327-330.
[22]由路.硒蛋白T与鸡缺硒性免疫功能障碍的关系[D].黑龙江哈尔滨:东北农业大学,2014.
[23]ZHANG W,ZHANG R,WANG T,et al.Selenium inhibits LPS-induced pro-inflammatory gene expression by modulating MAPK and NF-κB signaling pathways in mouse mammary epithelial cells in primary culture[J].Inflammation,2014,37(2):478-485.