饲粮中添加共轭亚油酸对猪肌肉组织miRNA表达谱的影响

doi:10.11843/j.issn.0366-6964.2018.09.012

Abstract

Abstract:

This study was conducted to investigate the molecular mechanisms of muscle metabolism regulated by conjugated linoleic acid (CLA) through affecting the expression of miRNA in porcine muscle. Twelve purebred Rongchang gilts with similar body weight were randomly divided into control and CLA groups. The pregnant gilts in CLA group were fed with 1.5% CLA diet from the beginning of pregnancy to weaning at 28 days old; After weaning, 30 piglets were respectively selected from the control and CLA groups, and the diet of piglets in original CLA group was also added 1.5% CLA. When the body weight of piglets was about 30 kg, 6 piglets from each group were slaughtered to collect the back muscle and leg muscle samples. After RNA extraction, cDNA libraries were constructed by the back muscle and leg muscle samples of the control and CLA groups; The Solexa deep sequencing was performed to analyze the effect of CLA on the miRNA expression profiles in porcine muscle tissue, the target genes prediction and function analysis for differentially expressed miRNA were carried out through the bioinformatics method. The results showed that:1)There were 44 869 982 and 45 105 806 clean reads in the back muscle and leg muscle tissues, respectively, and most of the small RNA sequence were 20-23 nt. 2) 306 and 304 known miRNAs were respectively found in the back muscle and leg muscle tissues, 295 known miRNAs were co-expressed in two tissues. 3) The expression of miRNA in porcine muscle was changed by CLA supplementation and the effect of CLA on miRNA expression in leg muscle was stronger than that in back muscle; 5 miRNAs in the back muscle and 12 miRNAs in the leg muscle were differentially expressed in corresponding CLA treatment (P<0.05), of which only the ssc-miR-224 was differentially expressed in both two tissues, thus 16 differentially expressed miRNAs were identified. 4) KEGG analysis revealed that the target genes of 16 differentially expressed miRNAs were involved in 292 biological pathways, of which 24 pathways were significantly enriched (P<0.05), including the MAPK and Notch signaling pathways, which were known to regulate muscle metabolism. 5) Six differentially expressed miRNAs were randomly selected to validate the sequencing results by qRT-PCR, the qRT-PCR expression results corresponded well with the results from the sequencing. This study suggests that CLA may regulate muscle metabolic pathways by affecting the expression of miRNA.

Key words: CLA, pig, miRNA, Solexa sequencing, muscle tissue

CLC Number:

S828.5

WANG Qi, QI Ren-li, LIU Hong, WANG Jing, HUANG Jin-xiu. Effects of Conjugated Linoleic Acid Supplementation in Diet on the Expression Profile of miRNAs in Porcine Muscle Tissue[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 1908-1918.

References

[1] LEHNEN T E,DA SILVA M R,CAMACHO A,et al.A review on effects of conjugated linoleic fatty acid (CLA) upon body composition and energetic metabolism[J]. J Int Soc Sports Nutr,2015,12:36.
[2] WANG Y W,JONES P J H.Dietary conjugated linoleic acid and body composition[J].Am J Clin Nutr,2004,79(6):1153S-1158S.
[3] FERLAY A,BERNARD L,MEYNADIER A,et al.Production of trans and conjugated fatty acids in dairy ruminants and their putative effects on human health:a review[J].Biochimie,2017,141:107-120.
[4] QI R L,YANG F Y,HUANG J X,et al.Supplementation with conjugated linoeic acid decreases pig back fat deposition by inducing adipocyte apoptosis[J].BMC Vet Res,2014,10:141.
[5] KIM Y,KIM J,WHANG K Y,et al.Impact of conjugated linoleic acid (CLA) on skeletal muscle metabolism[J].Lipids,2016,51(2):159-178.
[6] 黄金秀, 杨飞云,刘作华,等. 共轭亚油酸对体外培养的猪骨骼肌肌纤维类型组成的影响[J]. 畜牧兽医学报,2010,41(3):295-300.
HUANG J X, YANG F Y, LIU Z H, et al. Effect of conjugated linoleic acid on the composition of myofiber types in skeletal muscle cells of pigs in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2010, 41(3):295-300. (in Chinese)
[7] VIENBERG S,GEIGER J,MADSEN S,et al.microRNAs in metabolism[J].Acta Physiol,2017,219(2):346-361.
[8] 盛熙晖, 邓桂馨, 倪和民, 等.microRNAs调控动物骨骼肌发育的研究进展[J]. 畜牧兽医学报, 2015, 46(2):179-185.
SHENG X H, DENG G X, NI H M, et al. Research progress on microRNAs regulating animal skeletal muscle development[J]. Acta Veterinaria et Zootechnica Sinica, 2015, 46(2):179-185. (in Chinese)
[9] COUTURIER A,KELLER J,MOST E,et al.Niacin in pharmacological doses alters microRNA expression in skeletal muscle of obese Zucker rats[J].PLoS One,2014,9(5):e98313.
[10] LI Y J,LI J L,ZHANG L,et al.Effects of dietary energy sources onPost mortem glycolysis,meat quality and muscle fibre type transformation of finishing pigs[J].PLoS One,2015,10(6):e0131958.
[11] PARRA P,SERRA F,PALOU A.Expression of adipose microRNAs is sensitive to dietary conjugated linoleic acid treatment in mice[J].PLoS One,2010,5(9):e13005.
[12] QI R L,CHEN Y,HUANG J X,et al.Effects of conjugated linoleic acid on the expression levels of miR-27 and miR-143 in pig adipose tissue[J].Genet Mol Res,2015,14(2):6985-6992.
[13] HUANG J X,QI R L,CHEN X L,et al.Improvement in the carcass traits and meat quality of growing-finishing Rongchang pigs by conjugated linoleic acid through altered gene expression of muscle fiber types[J].Genet Mol Res,2014,13(3):7061-7069.
[14] 王琪,齐仁立,王敬,等.从胚胎期到育肥期饲粮添加共轭亚油酸对猪肉品质、脏器指数及脂肪酸组成的影响[J].中国畜牧杂志,2017,53(2):79-84.
WANG Q,QI R L,WANG J,et al.Effects of CLA supplementation on meat quality,organ index and fatty acid composition of pigs from embryo to finishing period[J].Chinese Journal of Animal Science,2017,53(2):79-84.(in Chinese)
[15] CORINO C,PASTORELLI G,ROSI F,et al.Effect of dietary conjugated linoleic acid supplementation in sows on performance and immunoglobulin concentration in piglets[J].J Anim Sci,2009,87(7):2299-2305.
[16] ZHOU L,CHEN J,LI Z,et al.Integrated profiling of microRNAs and mRNAs:microRNAs located on Xq27.3 associate with clear cell renal cell carcinoma[J].PLoS One,2010,5(12):e15224.
[17] FRONTERA W R,OCHALA J.Skeletal muscle:a brief review of structure and function[J].Calcif Tissue Int,2015,96(3):183-195.
[18] 吴望军,陈杰,黄瑞华,等.高通量技术挖掘猪microRNA的研究进展[J].畜牧兽医学报,2013,44(12):1857-1866.
WU W J,CHEN J,HUANG R H,et al.Advances in porcine microRNA research using high-throughput technologies[J].Acta Veterinaria et Zootechnica Sinica,2013, 44(12):1857-1866.(in Chinese)
[19] GE Y J,CHEN J.microRNAs in skeletal myogenesis[J].Cell Cycle,2011,10(3):441-448.
[20] QIN L J,CHEN Y S,LIU X H,et al.Integrative analysis of porcine microRNAome during skeletal muscle development[J].PLoS One,2013,8(9):e72418.
[21] JANSEN F,STUMPF T,PROEBSTING S,et al.Intercellular transfer of miR-126-3p by endothelial microparticles reduces vascular smooth muscle cell proliferation and limits neointima formation by inhibiting LRP6[J].J Mol Cell Cardiol,2017,104:43-52.
[22] HUANG S C,WANG M,WU W B,et al.Mir-22-3p Inhibits arterial smooth muscle cell proliferation and migration and neointimal hyperplasia by targeting HMGB1 in arteriosclerosis obliterans[J].Cell Physiol Biochem,2017,42(6):2492-2506.
[23] PENG Y D,XIANG H,CHEN C,et al.MiR-224 impairs adipocyte early differentiation and regulates fatty acid metabolism[J].Int J Biochem Cell Biol,2013, 45(8):1585-1593.
[24] STACHOWIAK M,SZYDLOWSKI M,FLISIKOWSKI K,et al.Polymorphism in 3' untranslated region of the pigPPARA gene influences its transcript level and is associated with adipose tissue accumulation[J].J Anim Sci,2014,92(6):2363-2371.
[25] WANG Z X,LI Q G,CHAMBA Y,et al.Identification of genes related to growth and lipid deposition from transcriptome profiles of pig muscle tissue[J].PLoS One, 2017, 12(2):e0172930.
[26] 陈晨,胡雄贵,朱吉,等.猪脂肪发育相关miRNAs的功能研究进展[J].畜牧兽医学报,2015,46(12):2117-2126.
CHEN C,HU X G,ZHU J,et al.Progress on the research of miRNAs associated with fat development in pigs[J].Acta Veterinaria et Zootechnica Sinica,2015,46(12):2117-2126.(in Chinese)
[27] CASAS-AGUSTENCH P,FERNANDES F S,DO CARMO M G T,et al.Consumption of distinct dietary lipids during early pregnancy differentially modulates the expression of microRNAs in mothers and offspring[J].PLoS One,2015,10(2):e0117858.
[28] JEONG B C,KANG I H,HWANG Y C,et al.MicroRNA-194 reciprocally stimulates osteogenesis and inhibits adipogenesis via regulating COUP-TFⅡ expression[J]. Cell Death Dis,2014,5:e1532.
[29] YU J W,CHEN Y S,QIN L M,et al.Effect of miR-205 on 3T3-L1 preadipocyte differentiation through targeting to glycogen synthase kinase 3 beta[J].Biotechnol Lett,2014,36(6):1233-1243.
[30] YU Y H,LI X,LIU L Y,et al.miR-628 promotes burn-induced skeletal muscle atrophy via targeting IRS1[J].Int J Biol Sci,2016,12(10):1213-1224.
[31] ZHANG Y,WANG R J,DU W J,et al.Downregulation of miR-151-5p contributes to increased susceptibility to arrhythmogenesis during myocardial infarction with estrogen deprivation[J].PLoS One,2013,8(9):e72985.
[32] HENDGEN-COTTA U B,MESSIHA D,ESFELD S,et al.Inorganic nitrite modulates miRNA signatures in acute myocardialin vivo ischemia/reperfusion[J].Free Radic Res,2017,51(1):91-102.
[33] SELIM K A,ABDELRASOUL H,ABOELMAGD M,et al.The role of the MAPK signaling,topoisomerase and dietary bioactives in controlling cancer incidence[J]. Diseases,2017,5(2):13.
[34] KYOSSEVA S V.Targeting MAPK signaling in age-related macular degeneration[J].Ophthalmol Eye Dis,2016,8:23-30.
[35] SEGALÉS J,PERDIGUERO E,MUÑOZ-CÁNOVES P.Regulation of muscle stem cell functions:a focus on the p38 MAPK signaling pathway[J].Front Cell Dev Biol,2016,4:91.
[36] QI R,LIU H,WANG Q,et al.Expressions and regulatory effects of P38/ERK/JNK Mapks in the adipogenic trans-differentiation of C2C12 myoblasts[J].Cell Physiol Biochem,2017,44(6):2467-2475.
[37] BRIEN P,PUGAZHENDHI D,WOODHOUSE S,et al.p38α MAPK regulates adult muscle stem cell fate by restricting progenitor proliferation during postnatal growth and repair[J].Stem Cells,2013,31(8):1597-1610.
[38] LEE J H,TACHIBANA H,MORINAGA Y,et al.Modulation of proliferation and differentiation of C2C12 skeletal muscle cells by fatty acids[J].Life Sci,2009,84(13-14):415-420.
[39] HUANG W C,TU R S,CHEN Y L,et al.Conjugated linoleic acids suppress inflammatory response and ICAM-1 expression through inhibition of NF-κB and MAPK signaling in human bronchial epithelial cells[J].Food Function,2016,7(4):2025-2033.
[40] BI P P,YUE F,SATO Y,et al.Stage-specific effects of Notch activation during skeletal myogenesis[J].eLife,2016,5:e17355.
[41] QIN L L,XU J,WU Z F,et al.Notch1-mediated signaling regulates proliferation of porcine satellite cells (PSCs)[J].Cell Signal,2013,25(2):561-569.
[42] PAJVANI U B,QIANG L,KANGSAMAKSIN T,et al.Inhibition of Notch uncouples Akt activation from hepatic lipid accumulation by decreasing mTorc1 stability[J]. Nat Med,2013,19(8):1054-1060.
[43] MASUDA S,TANAKA M,INOUE T,et al.Chemokine (C-X-C motif) ligand 1 is a myokine induced by palmitate and is required for myogenesis in mouse satellite cells[J].Acta Physiol,2017,222(3):e12975.

Effects of Conjugated Linoleic Acid Supplementation in Diet on the Expression Profile of miRNAs in Porcine Muscle Tissue

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