长途运输影响安格斯育肥牛肝脏功能关键基因的筛选

doi:10.16431/j.cnki.1671-7236.2023.10.023

摘要/Abstract

摘要： 【目的】研究长途运输对育肥牛体重、屠宰率及肝脏mRNA表达谱的影响,筛选长途运输影响肝脏功能的关键基因。【方法】选用体重相近的安格斯育肥公牛22头,随机分为2组,对照组9头就地屠宰,试验组13头经长途运输后屠宰,观察长途运输对体重和屠宰率的影响;此外,两组分别采集3头体重相近牛的肝脏组织,进行转录组测序(RNA-Seq),筛选差异表达基因,并对其进行GO功能和KEGG通路富集分析。【结果】长途运输后牛活体重降低4.5%,屠宰率较对照组无显著差异(P＞0.05);RNA-Seq结果发现,长途运输组肝脏组织有182个基因显著上调表达,91个基因显著下调表达。GO功能与KEGG通路富集分析显示,182个上调表达基因显著富集在刺激反应的调节、过氧化物酶体和氧化还原酶活性,以及cAMP、MAPK和长寿调控通路等,与能量代谢及肝细胞损伤密切相关。根据GO功能与KEGG通路富集分析结果,筛选出4个与育肥牛长途运输后肝脏能量代谢变化、肝脏细胞损伤及损伤后适应性保护机制密切相关的基因:ACOX3、FOS、HSPA1A、HSPA6。【结论】安格斯育肥牛经长途运输后,活体重较运输前降低4.5%,筛选得到4个长途运输影响安格斯育肥牛肝脏功能的关键基因:ACOX3、FOS、HSPA1A与HSPA6。研究结果为今后肉牛运输应激发生机理、预防和治疗研究提供参考。

关键词: 长途运输; 屠宰率; 肝脏功能; 肉牛

Abstract: 【Objective】 This experiment was conducted to study the effects of long-distance transportation on body weight,slaughter rate and liver mRNA expression profile of fattening cattle and to screen for key genes affecting liver function in long-distance transportation.【Method】 Twenty-two Angus fattening bulls of similar weight were randomly divided into 2 groups,9 of which were slaughtered on the spot in the control group,and 13 of which were slaughtered after long-distance transportation in the experimental group,and the effect of long-distance transportation on body weight and slaughter rate was observed.In addition,three bovine liver tissues of similar weight were collected in the two groups.Transcriptome sequencing (RNA-Seq) was performed to screen differentially expressed genes,and GO functional annotation and KEGG pathway enrichment analysis were performed on them.【Result】 The live body weight of cattle decreased by 4.5% after long-distance transportation,and there was no significant difference in slaughter rate compared with the control group (P＞0.05).The RNA-Seq results showed that 182 genes were significantly upregulated and 91 genes were significantly downregulated in liver tissues of the long-distance transportation group.GO enrichment and KEGG pathway analysis showed that 182 up-regulated genes were significantly enriched in the terms of regulation of response to stimulus,peroxisome,oxidoreductase activity,cAMP signaling pathway,MAPK signaling pathway,and longevity regulating pathway.According to the results of GO enrichment and KEGG pathway analysis,four genes (ACOX3,FOS, HSPA1A,HSPA6) were screened out that were closely related to the changes of liver energy metabolism,liver cell damage and post-injury adaptive protection mechanism of fattening cattle after long-distance transportation.【Conclusion】 The live weight of Angus fattening cattle was reduced by 4.5% after long-distance transportation.Four key genes ACOX3,FOS,HSPA1A and HSPA6 affecting liver function of Angus fattening cattle after long-distance transportation were screened.The results provided a reference for future research on the mechanism,prevention and treatment of transport stress in beef cattle.

Key words: long-distance transportation; slaughter rate; liver function; beef cattle

中图分类号:

S811.2

孙文阳, 谢建亮, 施安, 陈志龙, 姜武, 王瑞刚, 侯鹏霞, 梁小军. 长途运输影响安格斯育肥牛肝脏功能关键基因的筛选[J]. 中国畜牧兽医, 2023, 50(10): 4096-4105.

SUN Wenyang, XIE Jianliang, SHI An, CHEN Zhilong, JIANG Wu, WANG Ruigang, HOU Pengxia, LIANG Xiaojun. Screening of Key Genes Affecting Liver Function in Angus Fattening Cattle After Long-distance Transportation[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 4096-4105.

参考文献

[1] VAN ENGEN N K,COETZEE J F.Effects of transportation on cattle health and production:A review[J].Animal Health Research Reviews,2018,19(2):142-154.
[2] DELIC N,ALEKSIC S,PETROVIC M M,et al.Methods for determining stress syndrome in beef cattle and its relevance to quality of meat[J].Biotechnology in Animal Husbandry,2014,30:37-44.
[3] GONZALEZ L A,SCHWARTZKOPF-GENSWEIN K S,BRYAN M,et al.Factors affecting body weight loss during commercial long haul transport of cattle in North America[J].Journal of Animal Science,2012,90(10):3630-3639.
[4] LITTLEJOHN B P,NEUENDORFF D A,CARROLL J A,et al.Effects of prenatal transportation stress on days of age at first calving in brahman heifers and their calves' growth and temperament at weaning[J].Journal of Animal Science,2016,95:44.
[5] GRUBER S,TATUM J,ENGLE T,et al.Relationships of behavioral and physiological symptoms of preslaughter stress to beef longissimus muscle tenderness[J].Journal of Animal Science,2010,88(3):1148-1159.
[6] 李妍,高艳霞,许利民,等.长途运输应激对西门塔尔牛体重、血液生化指标和细胞因子含量的影响[J].中国畜牧兽医,2018,45(5):1234-1240. LI Y,GAO Y X,XU L M,et al.Effect of long-distance transportation stress on the body weight,blood biochemical indexes and cytokine contents of Simmental cattle[J].China Animal Husbandry & Veterinary Medicine,2018,45(5):1234-1240.(in Chinese)
[7] 芦春莲,李妍,曹玉凤,等.肉牛宰前运输应激对其血液理化指标及免疫机能的影响[J].中国兽医学报,2016,36(7):1173-1177. LU C L,LI Y,CAO Y F,et al.Effects of preslaughter transportation stress on blood physiological and biochemical indexes and immunity function of beef cattle[J].Chinese Journal of Veterinary Science,2016,36(7):1173-1177.(in Chinese)
[8] 许利凡,贺丛,王绿林,等.运输应激对夏南牛血清促肾上腺皮质激素、皮质醇、白介素-6、肿瘤坏死因子-α水平的影响[J].黑龙江畜牧兽医,2014,15:88-90. XU L F,HE C,WANG L L,et al.Effects of transportation stress on the level serum adrenocortical hormone,cortisol,interleukin-6 and tumor necrosis factor-alpha in Xianan cattle[J].Heilongjiang Animal Science and Veterinary Medicine,2014,15:88-90.(in Chinese)
[9] 刘延鑫,王义翠,孙宇,等.运输应激牛不同组织HSPs mRNA和蛋白表达研究[J].黑龙江畜牧兽医,2018,13:75-78. LIU Y X,WANG Y C,SUN Y,et al.Study on the HSPs mRNA and protein expression of different tissues in transport stressed beef cattle[J].Heilongjiang Animal Science and Veterinary Medicine,2018,13:75-78.(in Chinese)
[10] 杨高丰.运输对夏南牛外周血淋巴细胞HSPs基因表达的影响及HSP70基因的克隆[D].郑州:河南农业大学,2012. YANG G F.Expression of HSPs in the peripheral blood lymphocytes of Xianan beef cattle by transportation durations and cloning of HSP70 gene[D].Zhengzhou:Henan Agricultural University,2012.(in Chinese)
[11] 马爱霞,齐艳君,陈凤梅,等."健牛散"对肉牛运输应激综合征的影响[J].动物医学进展,2021,42(12):61-65. MA A X,QI Y J,CHEN F M,et al.The effect of "Jianniusan" on beef cattle transport stress syndrome[J].Progress in Veterinary Medicine,2021,42(12):61-65.(in Chinese)
[12] ZHAO H D,TANG X Q,WU M L,et al.Transcriptome characterization of short distance transport stress in beef cattle blood[J].Frontiers in Genetics,2021,12:616388.
[13] TITORENKO V I,RACHUBINSKI R A.Dynamics of peroxisome assembly and function[J].Trends in Cell Biology,2001,11(1):22-29.
[14] MI J,KIRCHNER E,CRISTOBAL S.Quantitative proteomic comparison of mouse peroxisomes from liver and kidney[J].Proteomics,2007,7(11):1916-1928.
[15] ISLINGER M,VOELKL A,FAHIMI H D.The peroxisome:An update on mysteries 2.0[J].Histochemistry and Cell Biology,2018,150(5):443-471.
[16] RAUCH N,RUKHLENKO O S,KOLCH W,et al.MAPK kinase signalling dynamics regulate cell fate decisions and drug resistance[J].Current Opinion in Structural Biology,2016,41:151-158.
[17] LI Z L,ZHAO Q W,LU Y J,et al.DDIT4 s-nitrosylation aids p38-MAPK signaling complex assembly to promote hepatic reactive oxygen species production[J].Advanced Science,2021,8(18):e2101957.
[18] FERDINANDUSSE S,DENIS S,VAN ROERMUND C W T,et al.A novel case of ACOX2 deficiency leads to recognition of a third human peroxisomal acyl-CoA oxidase[J].Biochimica et Biophysica Acta- Molecular Basis of Disease,2018,1864(3):952-958.
[19] CHINENOV Y,KERPPOLA T K.Close encounters of many kinds:Fos-Jun interactions that mediate transcription regulatory specificity[J].Oncogene,2001,20(19):2438-2452.
[20] BAKIRI L,HAMACHER R,GRANA O,et al.Liver carcinogenesis by FOS-dependent inflammation and cholesterol dysregulation[J].Journal of Experimental Medicine,2017,214(5):1387-1409.
[21] SHAN W,QING P,WEI N,et al.HSPA1A protects cells from thermal stress by impeding ESCRT-0-mediated autophagic flux in epidermal thermoresistance[J].Journal of Investigative Dermatology,2021,141(1):48-58.
[22] ROSENZWEIG R,NILLEGODA N B,MAYER M P,et al.The Hsp70 chaperone network[J].Nature Reviews Molecular Cell Biology,2019,20(11):665-680.
[23] FERNANDEZ-FERNANDEZ M R,GRAGERA M,OCHOA-IBARROLA L,et al.Hsp70-a master regulator in protein degradation[J].FEBS Letters,2017,591(17):2648-2660.
[24] QU B G,JIA Y G,LIU Y C,et al.The detection and role of heat shock protein 70 in various nondisease conditions and disease conditions:A literature review[J].Cell Stress Chaperones,2015,20(6):885-892.
[25] KUBOKI S,SCHUSTER R,BLANCHARD J,et al.Role of heat shock protein 70 in hepatic ischemia-reperfusion injury in mice[J].American Journal of Physiology-Gastrointestinal and Liver Physiology,2007,292(4):G1141-1149.
[26] LI L B,ZHANG T R,LI Z,et al.Schisandrin B attenuates acetaminophen-induced hepatic injury through heat-shock protein 27 and 70 in mice[J].Journal of Gastroenterology and Hepatology,2014,29(3):640-647.
[27] LIU J L,DU S Y,KONG Q Y,et al.HSPA12A attenuates lipopolysaccharide-induced liver injury through inhibiting caspase-11-mediated hepatocyte pyroptosis via PGC-1α-dependent acyloxyacyl hydrolase expression[J].Cell Death and Differentiation,2020,27(9):2651-2667.
[28] LIU Y H,YU M J,CUI J W,et al.Heat shock proteins took part in oxidative stress-mediated inflammatory injury via NF-κB pathway in excess manganese-treated chicken livers[J].Ecotoxicology and Environmental Safety,2021,226:112833.
[29] GOMEZ-PASTOR R,BURCHFIEL E T,THIELE D J.Regulation of heat shock transcription factors and their roles in physiology and disease[J].Nature Reviews Molecular Cell Biology,2018,19(1):4-19.
[30] BI J B,ZHANG J,KE M Y,et al.HSF2BP protects against acute liver injury by regulating HSF2/HSP70/MAPK signaling in mice[J].Cell Death and Disease,2022,13(9):830.