徐鑫;黄兴法;王秀英;汪文俊;朱惠玲;刘玉兰;张晶;
XU Xin;HUANG Xingfa;WANG Xiuying;WANG Wenjun;ZHU Huiling;LIU Yulan;ZHANG Jing;Hubei Key Laboratory of Animal Nutrition and Feed Science, School of Animal Science and Nutritional Engineering, Wuhan Polytechnic University;Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China, College of Life Sciences, South-Central University for Nationalities;

• 1:武汉轻工大学动物科学与营养工程学院动物营养与饲料科学湖北省重点实验室
• 2:中南民族大学生命科学学院武陵山区特色资源植物种质保护与利用湖北省重点实验室

摘要(Abstract)：

本试验旨在研究脂多糖(LPS)诱导仔猪发生炎症反应时肝脏和脾脏炎性细胞因子和lncRNA-47491、lncRNA-49770、lncRNA-51636表达的变化。选取8头21日龄、体重(7.15±0.42)kg的杜×长×大三元杂交断奶仔猪,随机分为对照组、LPS组,每个处理4个重复,每个重复1头猪。预试14 d后,LPS组和对照组腹腔分别注射100μg/kg体重的LPS、生理盐水,4 h后屠宰并取肝脏和脾脏组织样待测。结果表明:与对照组相比,LPS刺激使肝脏和脾脏中炎性细胞因子的mRNA表达量显著上调;LPS诱导的炎症反应中lncRNA-47491、lncRNA-49770和lncRNA-51636的表达量均显著上调;对lncRNA调控的顺式靶标基因的预测结果显示,lncRNA-47491和lncRNA-51636的靶基因有TMEM235、PGS1、TK1等,lncRNA-49770的靶基因有SRRM2、CLDN6、TNFRSF12A等,且这些基因均与炎症相关。综上,lncRNA-47491、lncRNA-49770及lncRNA-51636可能参与了机体的炎症反应,且靶标基因的预测提示其可能发挥不同的调控作用。
The purpose of this study was to investigate the changes in the expression of inflammatory cytokines and lncRNA-47491, lncRNA-49770, and lncRNA-51636 in liver and spleen induced by lipopolysaccharide(LPS) in piglets. A total of 8 weaned piglets(Duroc × Landrace × Yorkshine; weaned at 21 d of age;(7.15±0.42) kg initial body weight) were randomly divided into two groups(n=4), including control group and LPS group. After 14 days of pretest, the piglets in LPS group were injected with 100 μg/kg BW LPS and the piglets in control group were injected with an equivalent amount of sterile physiological saline solution. All piglets were slaughtered at 4 h post injections and tissue samples of liver and spleen were collected for analysis. The results showed that in liver and spleen tissues, LPS significantly increased the expression of inflammatory cytokines and lncRNA(lncRNA-47491, lncRNA-49770 and lncRNA-51636). Target gene prediction results show that the potential target genes of lncRNA-47491 and lncRNA-51636 including TMEM235, PGS1, TK1, target genes of lncRNA-49770 including SRRM2, CLDN6, TNFRSF12 A, and these genes are all associated with inflammation. Therefore, lncRNA-47491, lncRNA-49770 and lncRNA-51636 may be involved in the inflammatory response, and the prediction of these target genes revealed that they may play different regulatory roles.

关键词(KeyWords)： lncRNA;脂多糖;肝脏;脾脏;断奶仔猪
lncRNA;Lipopolysaccharide;Liver;Spleen;Weaned piglets

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31702203);; 湖北省教育厅优秀中青年科技创新团队项目(T201508)

作者(Authors): 徐鑫;黄兴法;王秀英;汪文俊;朱惠玲;刘玉兰;张晶;
XU Xin;HUANG Xingfa;WANG Xiuying;WANG Wenjun;ZHU Huiling;LIU Yulan;ZHANG Jing;Hubei Key Laboratory of Animal Nutrition and Feed Science, School of Animal Science and Nutritional Engineering, Wuhan Polytechnic University;Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China, College of Life Sciences, South-Central University for Nationalities;

DOI: 10.19556/j.0258-7033.20190704-03

参考文献(References)：

• [1]Sabroe I, Parker L C, Dower S K, et al. The role of TLR activation in inflammation[J]. J Pathology, 2008, 214(2):126-135.
• [2]Kornienko A E, Guenzl P M, Barlow D P, et al. Gene regulation by the act of long non-coding RNA transcription[J]. BMC Biol,2013, 11(1):59.
• [3]Dai L, Zhang G, Cheng Z, et al. Knockdown of LncRNA MALAT1contributes to the suppression of inflammatory responses by upregulating miR-146a in LPS-induced acute lung injury[J]. Connect Tissue Res, 2018, 59(6):581-592.
• [4]Du M, Yuan L, Tan X, et al. The LPS-inducible lncRNA Mirt2is a negative regulator of inflammation[J]. Nat Commun, 2017,8(1):2049.
• [5]Wu H, Liu J, Li W, et al. LncRNA-HOTAIR promotes TNF-αproduction in cardiomyocytes of LPS-induced sepsis mice by activating NF-κB pathway[J]. Biochem Biophys Res Commun,2016, 471(1):240-246.
• [6]Liu Y, Chen F, Odle J, et al. Fish oil enhances intestinal integrity and inhibits TLR4 and NOD2 signaling pathways in weaned pigs after LPS challenge[J]. J Nutr, 2012, 142(11):2017-2024.
• [7]Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and 2-ΔΔCt method[J]. Methods, 2001, 25(4):402-408.
• [8]Leng W, Liu Y, Shi H, et al. Aspartate alleviates liver injury and regulates mRNA expressions of TLR4 and NOD signaling—related genes in weaned pigs after lipopolysaccharide challenge[J]. J Nutr Biochem, 2014, 25(6):592-599.
• [9]Xu L, Kwak M, Zhang W, et al. Time-dependent effect of E.coli LPS in spleen DC activation in vivo:alteration of numbers,expression of co-stimulatory molecules, production of proinflammatory cytokines, and presentation of antigens[J]. Mol Immunol, 2017, 85:205-213.
• [10]Yang G, Lu X, Yuan L. LncRNA:A link between RNA and cancer[J]. Biochim Biophys Acta, 2014, 1839(11):1097-1109.
• [11]Tan H T T, Hagner S, Ruchti F, et al. Tight junction, mucin, and inflammasome-related molecules are differentially expressed in eosinophilic, mixed, and neutrophilic experimental asthma in mice[J]. Allergy, 2019, 74(2):294-307.
• [12]郑文伟,何晓娟,贾良良,等.跳骨片含药血清抑制脂多糖诱导的软骨细胞炎症反应的作用机制研究[J].中医正骨,2017, 29(8):8-16.
• [13]孙悝,吕小艳,刘小云,等.呼吸系统炎症与肿瘤病人血清细胞质胸苷激酶1水平比较[J].蚌埠医学院学报, 2017,42(6):814-815.
• [14]Wang C, Zheng X, Shen C. MicroRNA-203 suppresses cell proliferation and migration by targeting BIRC5 and LASP1 in human triple-negative breast cancer cells[J]. J Exp Clin Cancer Res, 2012, 31(1):58.
• [15]Ungvári I, Hadadié, Virág V, et al. Implication of BIRC5 in asthma pathogenesis[J]. Int Immunol, 2012, 24(5):293-301.
• [16]Pi?eiro-Hermida S, López I P, Alfaro-Arnedo E, et al. IGF1R deficiency attenuates acute inflammatory response in a bleomycininduced lung injury mouse model[J]. Sci Rep, 2017, 7(1):4290.
• [17]Lewis J, Milner D, Lewis A, et al. Up-regulation of claudin-6in the distal lung impacts secondhand smoke-induced inflammation[J]. Int J Environ Res Public Health, 2016, 13(10):1018.
• [18]吴永刚,洪礼传,冯洁,等. CLDN9基因与垂体瘤样细胞瘤侵袭相关性的实验研究[J].中华神经外科杂志, 2016,32(12):1278-1282.
• [19]Liu K, Wang X J, Li Y N, et al. Tongxinluo reverses the hypoxiasuppressed claudin-9 in cardiac microvascular endothelial cells[J]. Chin Med J(Engl), 2016, 129(4):442-447.
• [20]Mykk?nen O T, Kirjavainen P V, Pulkkinen L, et al. Reduction of subacute endotoxemia could mediate beneficial effects of vaccinium myrtillus in metabolic syndrome-a pbmc gene expression study[J]. Acta Horticulturae, 2014(1017):369-379.
• [21]Van Kirk C A, Van Guilder H D, Young M, et al. Agerelated alterations in retinal neurovascular and inflammatory transcripts[J]. Mol Vis, 2011, 17(142):1261-1274.