强冷应激对阿勒泰羔羊血清IgG、皮质醇及小肠免疫相关细胞的影响
|
摘要
为了探讨较强寒冷刺激对阿勒泰羔羊血清IgG和皮质醇含量的影响及小肠黏膜免疫屏障的适应特征。把阿勒泰羔羊置于-30～-35℃分别进行0,6 h、1,4,12,24 d的冷刺激,利用酶联免疫吸附试验(ELISA)对阿勒泰羔羊血清中IgG和皮质醇的含量进行测定;通过石蜡切片和HE染色对阿勒泰羔羊小肠的显微结构进行观察;并通过改良甲苯胺蓝染色鉴定小肠中的甲苯胺蓝染色阳性细胞。发现冷应激后IgG质量浓度都比正常组明显降低,其中冷应激4 d与正常组质量浓度差异极显著(P<0.01),冷应激后皮质醇质量浓度比正常组表达上升,其中冷应激6 h时与正常组差异显著(P<0.05),而24 d与正常组差异极显著(P<0.01);IgG与皮质醇浓度在4,12和24 d呈较强正相关,6 h和1 d呈负相关,说明在强冷应激时,绵羊机体免疫机能的调节及抗炎症反应等受到了抑制。冷应激后十二指肠、空肠和回肠的显微结构发生了相应变化,其中最明显的是肠绒毛变短且变粗,回肠内淋巴小结数量增多;对小肠免疫相关细胞而言,同一温度下各肠段中细胞数量呈趋势变化,上皮内淋巴细胞和杯状细胞的数量在十二指肠、空肠和回肠中呈上升趋势,而肥大细胞数量呈下降趋势;冷应激后淋巴细胞数量较常温有所减少,杯状细胞和肥大细胞数量较常温下都有所增加,说明在强冷应激下机体通过激素的释放努力适应环境变化的同时,也会使小肠免疫功能下降。
In order to investigate the effect of strong cold stimulation on serum IgG and cortisol content in Altay lambs,and the adaptation characteristics of intestinal mucosal immune barriers,the Altay lambs were cold-stimulated for 0,6 h,1,4,12 and 24 d from-30℃ to-35℃.The content of IgG and cortisol in the serum were detected by enzyme-linked immunoabsorbent assay(ELISA).Paraffin section and HE staining was applied to ovserve the structure of the small intestine.The special positive cells in the small intestine were identified by a modified toluidine blue staining method.The IgG concentration after cold stress was lower than that of room temperature(P<0.01),while the cortisol concentration was higher than that of room temperature,with a significant difference at 6 h and(P<0.05),24 d(P<0.01);The content of IgG and cortisol showed a strong positive correlation at 4,12 and 24 d,and a negative correlation at 6 h and 1 d.The results showed that under cold stress,sheep immune function regulation and anti-inflammatory response were inhibited.The microstructure of the duodenum,jejunum,and ileum after cold stress displayed the changes,with the most obvious being the shortening and thickening of the intestinal villi,and the increased number of lymphoid nodules in the ileum;under the same temperature,the of small intestine-immune related cells showed a trend,of wich the number of lymph cells and goblet cells increased in the duodenum,jejunum,and ileum,while the number of mast cells showed a decreasing;after cold stress,the number of lymphocytes decreased compared with normal temperature,and the number of goblet cells and mast cells was slightly increased.This results indicated that the lambs tried to adapt the environmental changes through hormone release under strong cold stress,and it will also make the intestinal immune function decreased.
In order to investigate the effect of strong cold stimulation on serum IgG and cortisol content in Altay lambs,and the adaptation characteristics of intestinal mucosal immune barriers,the Altay lambs were cold-stimulated for 0,6 h,1,4,12 and 24 d from-30℃ to-35℃.The content of IgG and cortisol in the serum were detected by enzyme-linked immunoabsorbent assay(ELISA).Paraffin section and HE staining was applied to ovserve the structure of the small intestine.The special positive cells in the small intestine were identified by a modified toluidine blue staining method.The IgG concentration after cold stress was lower than that of room temperature(P<0.01),while the cortisol concentration was higher than that of room temperature,with a significant difference at 6 h and(P<0.05),24 d(P<0.01);The content of IgG and cortisol showed a strong positive correlation at 4,12 and 24 d,and a negative correlation at 6 h and 1 d.The results showed that under cold stress,sheep immune function regulation and anti-inflammatory response were inhibited.The microstructure of the duodenum,jejunum,and ileum after cold stress displayed the changes,with the most obvious being the shortening and thickening of the intestinal villi,and the increased number of lymphoid nodules in the ileum;under the same temperature,the of small intestine-immune related cells showed a trend,of wich the number of lymph cells and goblet cells increased in the duodenum,jejunum,and ileum,while the number of mast cells showed a decreasing;after cold stress,the number of lymphocytes decreased compared with normal temperature,and the number of goblet cells and mast cells was slightly increased.This results indicated that the lambs tried to adapt the environmental changes through hormone release under strong cold stress,and it will also make the intestinal immune function decreased.
引文
[1]DHANALAKSHMI S,DEVI R S,SRIKUMAR R,et al.Protective effect of triphala on cold stress-induced behavioral and biochemical abnormalities in rats[J].Yakugaku Zasshi,2007,127(11):1863-1867.
[2]HANGALAPURA B,NIEUWLAND M,BUYSE J,et al.Effect of duration of cold stress on plasma adrenal and thyroid hormone levels and immune responses in chicken lines divergently selected for antibody responses[J].Poul Sci,2004,83(10):1644-1649.
[3]吴永魁,计红,李乾学,等.冷应激对断乳仔猪ACTH及皮质醇的影响[J].应用与环境生物学报,2005,11(4):477-478.
[4]邵钺馨,单春花,王超,等.奶牛生产中冷热应激的研究进展[J].黑龙江畜牧兽医,2018,6(7):38-41.
[5]李建锋.不同放牧季节藏绵羊肠道免疫调节及组织形态学变化的研究[D].青海西宁:青海大学,2013.
[6]薛华山.阿勒泰羊品种资源现状及发展思路[J].当代畜牧,2016(5):39.
[7]马兰,常兰.不同海拔藏绵羊小肠上皮内淋巴细胞,杯状细胞的数量和分布比较及其意义[J].青海医学院学报,2012,33(2):99-102.
[8]张晖,肖红,张家禾,等.宁都黄鸡小肠显微结构和甲苯胺蓝染色阳性细胞分布特点[J].中国兽医学报,2013,33(7):1064-1068.
[9]MA P,CˇERVINKOVM,FUNDA D,et al.Enhancement by mild cold stress of the antibody forming capacity in euthymic and athymic hairless mice[J].Folia Microbiol,1998,43(5):521-523.
[10]刘宇,马红,汪亮,等.肾上腺素对猪季节性基因表达的影响[J].吉林农业大学学报,2017,39(3):322-325.
[11]姜冬梅,李士泽,杨焕民,等.冷应激家兔血压与血清细胞因子的变化及其相互关系[J].环境与健康杂志,2007(8):577-580.
[12]刘璐.冷应激对饥饿断奶羔羊生理生化指标及内分泌的影响[D].新疆乌鲁木齐:新疆农业大学,2014.
[13]黄丽波,李双武,孙牧文,等.不同强度冷应激对白凤雏鸡血清生化指标的影响[J].天津农学院学报,2017,24(3):59-63.
[14]李玮,刘蕤,马尧,等.重度冷应激对三河牛血液生化指标及相关基因表达的影响[J].畜牧兽医学报,2015,46(8):1463-1470.
[15]白丹丹,敖日格乐,王纯洁,等.慢性冷热应激对三河牛血液生化指标及相关基因表达的影响[J].中国农业大学学报,2017,22(8):50-56.
[16]CARR D J,WOOLLEY T W,BLALOCK J E.Phentolamine but not propranolol blocks the immunopotentiating effect of cold stress on antigen-specific igm production in mice orally immunized with sheep red blood cells[J].Brain Behav Immun,1992,6(1):50-63.
[17]SAPOLSKY R M,ROMERO L M,MUNCK A U.How do glucocorticoids influence stress responses?integrating permissive,suppressive,stimulatory,and preparative actions[J].Endocrine Rev,2000,21(1):55-89.
[18]KASPER L H,BUZONI-GATEL D.Ups and downs of mucosal cellular immunity against protozoan parasites[J].Infect Immun,2001,69(1):1-8.
[19]JOHNSON C,KUDSK K.Nutrition and intestinal mucosal immunity[J].Clin Nutr,1999,18(6):337-344
[20]胡艳艳,刘小伟.肠道杯状细胞结构和功能的研究进展[J].临床与病理杂志,2015,33(5):424-430.
[21]姜楠,胡易,陈耀星,等.肥大细胞在动物肠道黏膜免疫屏障中的作用[J].中国畜牧兽医,2011,38(8):167-170.
[22]左愈臻.牦牛小肠结构特点及黏膜免疫功能细胞分布变化研究[D].甘肃兰州:甘肃大学,2011.
[23]张鹏飞.藏羚羊小肠黏膜结构,黏膜免疫相关细胞和5-羟色胺免疫反应细胞的研究[D].青海西宁:青海大学,2012.
[2]HANGALAPURA B,NIEUWLAND M,BUYSE J,et al.Effect of duration of cold stress on plasma adrenal and thyroid hormone levels and immune responses in chicken lines divergently selected for antibody responses[J].Poul Sci,2004,83(10):1644-1649.
[3]吴永魁,计红,李乾学,等.冷应激对断乳仔猪ACTH及皮质醇的影响[J].应用与环境生物学报,2005,11(4):477-478.
[4]邵钺馨,单春花,王超,等.奶牛生产中冷热应激的研究进展[J].黑龙江畜牧兽医,2018,6(7):38-41.
[5]李建锋.不同放牧季节藏绵羊肠道免疫调节及组织形态学变化的研究[D].青海西宁:青海大学,2013.
[6]薛华山.阿勒泰羊品种资源现状及发展思路[J].当代畜牧,2016(5):39.
[7]马兰,常兰.不同海拔藏绵羊小肠上皮内淋巴细胞,杯状细胞的数量和分布比较及其意义[J].青海医学院学报,2012,33(2):99-102.
[8]张晖,肖红,张家禾,等.宁都黄鸡小肠显微结构和甲苯胺蓝染色阳性细胞分布特点[J].中国兽医学报,2013,33(7):1064-1068.
[9]MA P,CˇERVINKOVM,FUNDA D,et al.Enhancement by mild cold stress of the antibody forming capacity in euthymic and athymic hairless mice[J].Folia Microbiol,1998,43(5):521-523.
[10]刘宇,马红,汪亮,等.肾上腺素对猪季节性基因表达的影响[J].吉林农业大学学报,2017,39(3):322-325.
[11]姜冬梅,李士泽,杨焕民,等.冷应激家兔血压与血清细胞因子的变化及其相互关系[J].环境与健康杂志,2007(8):577-580.
[12]刘璐.冷应激对饥饿断奶羔羊生理生化指标及内分泌的影响[D].新疆乌鲁木齐:新疆农业大学,2014.
[13]黄丽波,李双武,孙牧文,等.不同强度冷应激对白凤雏鸡血清生化指标的影响[J].天津农学院学报,2017,24(3):59-63.
[14]李玮,刘蕤,马尧,等.重度冷应激对三河牛血液生化指标及相关基因表达的影响[J].畜牧兽医学报,2015,46(8):1463-1470.
[15]白丹丹,敖日格乐,王纯洁,等.慢性冷热应激对三河牛血液生化指标及相关基因表达的影响[J].中国农业大学学报,2017,22(8):50-56.
[16]CARR D J,WOOLLEY T W,BLALOCK J E.Phentolamine but not propranolol blocks the immunopotentiating effect of cold stress on antigen-specific igm production in mice orally immunized with sheep red blood cells[J].Brain Behav Immun,1992,6(1):50-63.
[17]SAPOLSKY R M,ROMERO L M,MUNCK A U.How do glucocorticoids influence stress responses?integrating permissive,suppressive,stimulatory,and preparative actions[J].Endocrine Rev,2000,21(1):55-89.
[18]KASPER L H,BUZONI-GATEL D.Ups and downs of mucosal cellular immunity against protozoan parasites[J].Infect Immun,2001,69(1):1-8.
[19]JOHNSON C,KUDSK K.Nutrition and intestinal mucosal immunity[J].Clin Nutr,1999,18(6):337-344
[20]胡艳艳,刘小伟.肠道杯状细胞结构和功能的研究进展[J].临床与病理杂志,2015,33(5):424-430.
[21]姜楠,胡易,陈耀星,等.肥大细胞在动物肠道黏膜免疫屏障中的作用[J].中国畜牧兽医,2011,38(8):167-170.
[22]左愈臻.牦牛小肠结构特点及黏膜免疫功能细胞分布变化研究[D].甘肃兰州:甘肃大学,2011.
[23]张鹏飞.藏羚羊小肠黏膜结构,黏膜免疫相关细胞和5-羟色胺免疫反应细胞的研究[D].青海西宁:青海大学,2012.