组胺对奶牛瘤胃上皮细胞mTOR自噬通路的影响
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摘要
奶牛发生亚急性瘤胃酸中毒时短链脂肪酸在瘤胃内大量积累,使瘤胃pH长时间为5. 0～5. 5,瘤胃内菌群紊乱,使组胺酸脱羧产生组胺的细菌增多,因而瘤胃内组胺浓度上升。瘤胃内自噬发生是清除异己,将体内的有害物排出以实现细胞本身的代谢需要和某些细胞器的更新,是机体的重要功能。试验研究了不同时间及浓度组胺对奶牛瘤胃上皮细胞自噬的影响(时间点0,1,3,6,9,18 h;浓度0. 8,1. 6,2. 4 nmol/m L)。结果表明:不同浓度组胺刺激后瘤胃上皮细胞自噬雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)通路中关键蛋白磷酸化mTOR(pmTOR)和自噬降解底物(p62)表达量上升,mTOR、微管相关蛋白1轻链3(LC3)表达量下降。说明组胺高浓度时mTOR自噬通路受到抑制,使清除有害物质功能减弱,从而引起细胞损伤。
When subacute rumen acidosis occurs in dairy cows,short-chain fatty acids accumulate in the rumen,causing rumen pH to be between 5. 0 and 5. 5 for a long time. The flora in the rumen is disordered,and the bacteria that deaminate histidine to produce histamine are increased,resulting in the increase of the concentration of histamine in the rumen. The autophagy in the rumen is the important function of the body to remove the dissidents and expel the harmful substances in the body to realize the metabolic needs of the cells themselves and the renewal of some organelles. This experiment was to investigate the effects of histamine at different times and concentrations on autophagy in rumen epithelial cells of dairy cows(time points 0,1,3,6,9 and 18 h; concentrations 0. 8,1. 6 and 2. 4 nmol/m L). The results showed that the expression of key protein phosphorylated mTOR(pmTOR) and autophagy degradation substrate(p62) increased in the rumen epithelial cells after stimulation with different concentrations of histamine,mTOR and mTOR. The expression level of microtubule-associated protein 1 light chain 3(LC3) was decreased. The results showed that the mTOR autophagy pathway was inhibited at high concentrations of histamine,which weakened the function of scavenging harmful substances and caused cell damage.
When subacute rumen acidosis occurs in dairy cows,short-chain fatty acids accumulate in the rumen,causing rumen pH to be between 5. 0 and 5. 5 for a long time. The flora in the rumen is disordered,and the bacteria that deaminate histidine to produce histamine are increased,resulting in the increase of the concentration of histamine in the rumen. The autophagy in the rumen is the important function of the body to remove the dissidents and expel the harmful substances in the body to realize the metabolic needs of the cells themselves and the renewal of some organelles. This experiment was to investigate the effects of histamine at different times and concentrations on autophagy in rumen epithelial cells of dairy cows(time points 0,1,3,6,9 and 18 h; concentrations 0. 8,1. 6 and 2. 4 nmol/m L). The results showed that the expression of key protein phosphorylated mTOR(pmTOR) and autophagy degradation substrate(p62) increased in the rumen epithelial cells after stimulation with different concentrations of histamine,mTOR and mTOR. The expression level of microtubule-associated protein 1 light chain 3(LC3) was decreased. The results showed that the mTOR autophagy pathway was inhibited at high concentrations of histamine,which weakened the function of scavenging harmful substances and caused cell damage.
引文
[1] Siciliano Jones J,Murphy M R.Production of volatile fatty acids in the rumen and cecum-colon of steers as affected by forage:concentrate and forage physical form 1[J]. J Dairy Sci,1989,72(2):485-492.
[2] Irwin L N,Mitchell G,Tucker R,et al. Histamine,tyramine,tryptamine and Electrolytes during glucose induced lactic acidosis[J]. Journal of Animal Science,1979,48(2):367-374.
[3]徐俊,苏衍菁,张养东,等.亚急性瘤胃酸中毒对瘤胃异常代谢产物和微生物组成的影响[J].中国奶牛,2012(9):17-20.
[4]王婷婷.亚急性瘤胃酸中毒病牛瘤胃组胺对瘤胃上皮细胞炎性通路的影响[D].长春:吉林大学,2015.
[5] Ahrens F A. Histamine,lactic acid,and hypertonicity as factors in the development of rumenitis in cattle[J]. American journal of veterinary research,1967,28(126):1335-1342.
[6] Ferstl R,Akdis C A,O'Mahony L. Histamine regulation of innate and adaptive immunity[J]. Frontiers in Bioscience(Landmark edition),2011,17:40-53.
[7]王之江.抑制组胺H3R受体可改善小鼠网膜下腔出血后早期脑损伤中的炎症反应:可能通过激活自噬抑制炎症[D].杭州:浙江大学,2017.
[8] Ding Suling,Abudupataer Mieradilijiang,Zhou Zheliang,et al.Histamine deficiency aggravates cardiac injury through miR-206/216b-Atg13 axis-mediated autophagic-dependant apoptosis[J]. Cell Death&Disease,2018,9(6):694.
[9] Zhang Z G,Li X B,Gao L,et al.An updated method for the isolation and culture of primary calf hepatocytes[J]. Vet J,2012,191(3):323-326.
[10] Grummer R R.Impact of changes in organic nutrient metabolism on feeding the transition cow[J]. Anim Sci,1995,73(9):2820-2833.
[11]贾哲,张才,吴庭才,等.奶牛脂肪肝细胞模型的建立[J].中国兽医学报,2018,38(5):1023-1028.
[12]舒适,肖鑫焕,王刚,等.奶牛亚临床低血钙症的蛋白质组学分析[J].中国兽医学报,2017,37(9):1756-1762.
[13]赵阳,钱伟东,曹宇,等.高产奶牛产后卵巢囊肿的血液临床病理学变化及其预警评估[J].中国兽医学报,2017,37(8):1600-1604.
[14]赖金伦,刘玉辉,陈颖钰,等.中药防治奶牛乳腺炎的临床应用及作用机制研究进展[J].中国兽医学报,2017,37(2):381-385.
[15] Drackley J R.Biology of dairy cows during the transition period:The final frontier[J]. Dairy Sci,1999,82:2259-2273.
[16]赵立香,宋晓静,郭梦尧,等.应用套式PCR-DGGE分析诊断奶牛产后急性子宫内膜炎致病菌[J].吉林农业大学学报,2013,35(5):591-596.
[17]姜明鑫,杨连玉.过瘤胃胆碱和蛋氨酸对泌乳早期奶牛生产性能和健康状况影响的研究现状[J].经济动物学报,2013,17(4):228-231,235.
[18]钱伟东,李昌盛,白云龙,等.高产奶牛产后卵巢静止血液临床病理学变化及其预警评估[J].中国兽医学报,2018,38(7):1400-1405.
[19] Akdis C A,Blaser K. Histamine in the immune regulation of allergic inflammation[J]. J Allergy Clin Immunol,2003,112(1):15-22.
[20]史楠,张燕,李晋峰,等.黄芪甲苷调控LC3/m TOR自噬通路在大鼠脑缺血再灌注损伤中的作用[J].检验医学与临床,2018,15(22):3363-3366.
[21] Kuma A,Hatano M,Matsui M,et al. The role of autophagy during the early neonatal starvation period[J]. Nature,2004432(7020):1032-1036.
[22] Yan Haijing,Zhang Xiangnan,Hu Weiwei,et al. Histamine H3 receptors aggravate cerebral ischaemic injury by histamine-independent mechanisms[J]. Nature Communications,2014,5:3334.
[2] Irwin L N,Mitchell G,Tucker R,et al. Histamine,tyramine,tryptamine and Electrolytes during glucose induced lactic acidosis[J]. Journal of Animal Science,1979,48(2):367-374.
[3]徐俊,苏衍菁,张养东,等.亚急性瘤胃酸中毒对瘤胃异常代谢产物和微生物组成的影响[J].中国奶牛,2012(9):17-20.
[4]王婷婷.亚急性瘤胃酸中毒病牛瘤胃组胺对瘤胃上皮细胞炎性通路的影响[D].长春:吉林大学,2015.
[5] Ahrens F A. Histamine,lactic acid,and hypertonicity as factors in the development of rumenitis in cattle[J]. American journal of veterinary research,1967,28(126):1335-1342.
[6] Ferstl R,Akdis C A,O'Mahony L. Histamine regulation of innate and adaptive immunity[J]. Frontiers in Bioscience(Landmark edition),2011,17:40-53.
[7]王之江.抑制组胺H3R受体可改善小鼠网膜下腔出血后早期脑损伤中的炎症反应:可能通过激活自噬抑制炎症[D].杭州:浙江大学,2017.
[8] Ding Suling,Abudupataer Mieradilijiang,Zhou Zheliang,et al.Histamine deficiency aggravates cardiac injury through miR-206/216b-Atg13 axis-mediated autophagic-dependant apoptosis[J]. Cell Death&Disease,2018,9(6):694.
[9] Zhang Z G,Li X B,Gao L,et al.An updated method for the isolation and culture of primary calf hepatocytes[J]. Vet J,2012,191(3):323-326.
[10] Grummer R R.Impact of changes in organic nutrient metabolism on feeding the transition cow[J]. Anim Sci,1995,73(9):2820-2833.
[11]贾哲,张才,吴庭才,等.奶牛脂肪肝细胞模型的建立[J].中国兽医学报,2018,38(5):1023-1028.
[12]舒适,肖鑫焕,王刚,等.奶牛亚临床低血钙症的蛋白质组学分析[J].中国兽医学报,2017,37(9):1756-1762.
[13]赵阳,钱伟东,曹宇,等.高产奶牛产后卵巢囊肿的血液临床病理学变化及其预警评估[J].中国兽医学报,2017,37(8):1600-1604.
[14]赖金伦,刘玉辉,陈颖钰,等.中药防治奶牛乳腺炎的临床应用及作用机制研究进展[J].中国兽医学报,2017,37(2):381-385.
[15] Drackley J R.Biology of dairy cows during the transition period:The final frontier[J]. Dairy Sci,1999,82:2259-2273.
[16]赵立香,宋晓静,郭梦尧,等.应用套式PCR-DGGE分析诊断奶牛产后急性子宫内膜炎致病菌[J].吉林农业大学学报,2013,35(5):591-596.
[17]姜明鑫,杨连玉.过瘤胃胆碱和蛋氨酸对泌乳早期奶牛生产性能和健康状况影响的研究现状[J].经济动物学报,2013,17(4):228-231,235.
[18]钱伟东,李昌盛,白云龙,等.高产奶牛产后卵巢静止血液临床病理学变化及其预警评估[J].中国兽医学报,2018,38(7):1400-1405.
[19] Akdis C A,Blaser K. Histamine in the immune regulation of allergic inflammation[J]. J Allergy Clin Immunol,2003,112(1):15-22.
[20]史楠,张燕,李晋峰,等.黄芪甲苷调控LC3/m TOR自噬通路在大鼠脑缺血再灌注损伤中的作用[J].检验医学与临床,2018,15(22):3363-3366.
[21] Kuma A,Hatano M,Matsui M,et al. The role of autophagy during the early neonatal starvation period[J]. Nature,2004432(7020):1032-1036.
[22] Yan Haijing,Zhang Xiangnan,Hu Weiwei,et al. Histamine H3 receptors aggravate cerebral ischaemic injury by histamine-independent mechanisms[J]. Nature Communications,2014,5:3334.