gga-miR-30a-5p的表达规律及其对鸡脂肪沉积的调控

doi:10.3864/j.issn.0578-1752.2025.15.014

摘要/Abstract

摘要：

【目的】 miR-30a-5p为miR-30家族成员，其在鸡脂肪沉积中的作用尚未见报道，通过对gga-miR-30a-5p在鸡腹部脂肪及肌内脂肪沉积中的作用研究，为解析鸡腹部脂肪和肌内脂肪生成的机制研究奠定基础。【方法】以优质黄羽肉鸡（矮小型）S3系和隐性白羽鸡（RR）为试验对象，利用荧光定量PCR检测gga-miR-30a-5p在腹部脂肪、肝脏和腿肌组织0周（0W）、2、4、8、14和16W及腹部脂肪细胞和肌内脂肪细胞增殖期、分化期的表达变化；腹部脂肪细胞及肌内脂肪细胞中分别转染gga-miR-30a-5p mimics和inhibitor，荧光定量PCR检测ga-miR-30a-5p的表达变化；油红O染色，异丙醇萃取脂滴，检测转染gga-miR-30a-5p mimics和inhibitor后腹部脂肪和肌内脂肪细胞脂滴沉积变化；利用生物信息学分析，预测gga-miR-30a-5p的作用靶基因。【结果】 gga-miR-30a-5p在不同组织（腹部脂肪、肝脏和腿肌组织）中的表达存在显著的品种差异性（P<0.05）。在腹部脂肪组织中，0周龄时gga-miR-30a-5p在隐性白羽鸡中的表达显著低于S3系鸡（P<0.05）。S3系和隐性白羽鸡0周龄时gga-miR-30a-5p表达最高，显著高于其他周龄（P<0.05），其他周龄间的表达差异不显著（P>0.05）；在肝脏组织中，16周龄时gga-miR-30a-5p在RR鸡中表达显著低于S3系鸡（P<0.05）。在S3系鸡中，16周龄肝脏组织中的gga-miR-30a-5p表达水平显著高于其他周龄（P<0.05），在RR鸡中，16周龄肝脏组织中的gga-miR-30a-5p表达水平显著高于0周龄和8周龄（P<0.05）；在腿肌组织中，16周龄时gga-miR-30a-5p在RR鸡中的表达显著低于S3系鸡（P<0.05）。gga-miR-30a-5p在S3系鸡14周龄时表达最低，显著低于2、8和16W（P<0.05），RR鸡腿肌组织各个周龄间的表达差异不显著（P>0.05）。在腹部脂肪细胞中，gga-miR-30a-5p在增殖期的表达显著低于分化期4 d和6 d的表达（P<0.05）；肌内脂肪细胞中，增殖期与分化期1 d的表达差异不显著（P>0.05），分化期4 d和6 d的表达水平显著高于增殖期（P<0.05），并且随着分化时间的延长，其表达水平逐渐升高；gga-miR-30a-5p mimics和inhibitor转染腹部脂肪细胞24 h后，gga-miR-30a-5p的表达显著升高和降低，表明gga-miR-30a-5p mimics和inhibitor已成功转染至腹部脂肪细胞；转染gga-miR-30a-5p mimics 3d后，腹部脂肪细胞脂滴沉积能力显著升高；转染gga-miR-30a-5p inhibitor，腹部脂肪细胞脂滴沉积能力则显著降低（P<0.05）；gga-miR-30a-5p mimics和inhibitor转染肌内脂肪细胞24 h后，gga-miR-30a-5p的表达显著升高和降低，表明gga-miR-30a-5p mimics和inhibitor已成功转染至肌内脂肪细胞；转染gga-miR-30a-5p mimics 3 d后，肌内脂肪细胞脂滴沉积能力显著升高；转染gga-miR-30a-5p inhibitor后，脂滴沉积能力则显著降低（P<0.05）。靶基因GO、Pathway及蛋白互作分析表明，UBE2I、UBE3C、CUL2、SOSC3和RUNX1可能是gga-miR-30a-5p调控脂肪沉积的预测靶基因。【结论】 gga-miR-30a-5p在不同组织中的表达存在显著的品种差异性；gga-miR-30a-5p具有促进腹部脂肪和肌内脂肪沉积的作用，UBE2I、UBE3C、CUL2、SOSC3和RUNX1是重要的候选靶基因。

关键词: 鸡, gga-miR-30a-5p, 腹部脂肪, 肌内脂肪

Abstract:

【Objective】 miR-30a-5p is a member of the miR-30 family, and its role in chicken fat deposition has not been reported. This study focused on the role of gga-miR-30a-5p in chicken abdominal fat and intramuscular fat deposition, which could lay a foundation for the analysis of the mechanism of chicken abdominal fat and intramuscular fat deposition. 【Method】 This study used high-quality yellow-feathered broilers (dwarf line, S3) and recessive white-feathered chickens (RR) as experimental subjects. Quantitative real-time PCR (qPCR) was employed to detect the expression changes of gga-miR-30a-5p in abdominal fat, liver, and leg muscle tissues at 0, 2, 4, 8, 14, and 16 weeks (0, 2, 4, 8, 14, and 16W), respectively, as well as during the proliferation and differentiation stages of abdominal and intramuscular adipocytes. Abdominal and intramuscular adipocytes were transfected with gga-miR-30a-5p mimics and inhibitors, and qPCR was used to measure the expression changes of gga-miR-30a-5p. Oil Red O staining and isopropanol extraction were performed to assess lipid droplet deposition after transfection. Bioinformatics analysis was conducted to predict the target genes of gga-miR-30a-5p. 【Result】 The expression of gga-miR-30a-5p in different tissues (abdominal fat, liver, and leg muscle) showed significant breed-specific differences (P<0.05). In abdominal fat tissue, the expression of gga-miR-30a-5p in recessive white-feathered chickens at 0W was significantly lower than that in S3 chickens (P<0.05). In both S3 and RR chickens, gga-miR-30a-5p expression in abdominal fat was the highest at 0W, significantly higher than that at other weeks (P<0.05), with no significant differences among other weeks (P>0.05). In liver tissue, gga-miR-30a-5p expression in RR chickens at 16W was significantly lower than that in S3 chickens (P<0.05). In S3 chickens, gga-miR-30a-5p expression in liver tissue at 16W was significantly higher than that at other weeks (P<0.05), while in RR chickens, its expression at 16W was significantly higher than that at 0 and 8W (P<0.05). In leg muscle tissue, gga-miR-30a-5p expression in RR chickens at 16W was significantly lower than that in S3 chickens (P<0.05). In S3 chickens, gga-miR-30a-5p expression in leg muscle was the lowest at 14W, significantly lower than that at 2, 8, and 16W (P<0.05), whereas no significant differences were observed among weeks in RR chickens (P>0.05). In abdominal adipocytes, gga-miR-30a-5p expression during the proliferation stage was significantly lower than that at differentiation days 4 and 6 (P<0.05). In intramuscular adipocytes, no significant difference was observed between the proliferation stage and differentiation day 1 (P>0.05), but expression at differentiation days 4 and 6 was significantly higher than during proliferation (P<0.05), gradually increasing with prolonged differentiation. After transfection with gga-miR-30a-5p mimics or inhibitors for 24 hours, gga-miR-30a-5p expression in abdominal adipocytes significantly increased or decreased, confirming successful transfection. Transfection with gga-miR-30a-5p mimics for 3 days significantly enhanced lipid droplet deposition in abdominal adipocytes, whereas transfection with inhibitors significantly reduced deposition (P<0.05). Similarly, in intramuscular adipocytes, transfection with mimics or inhibitors for 24 hours significantly altered gga-miR-30a-5p expression. Mimics transfection for 3 days significantly increased lipid droplet deposition, while inhibitor transfection significantly decreased it (P<0.05). Target gene prediction via GO, Pathway, and protein-protein interaction analyses suggested that UBE2I, UBE3C, CUL2, SOSC3, and RUNX1 were potential target genes of gga-miR-30a-5p in regulating fat deposition.【Conclusion】 gga-miR-30a-5p exhibited significant breed-specific expression differences across tissues, which promoted abdominal and intramuscular fat deposition, with UBE2I, UBE3C, CUL2, SOSC3, and RUNX1 identified as key candidate target genes.

Key words: chicken, gga-miR-30a-5p, abdominal fat, intramuscular fat

黄华云, 刘星, 王钱保, 李瑞瑞, 杨苗苗, 李春苗, 吴兆林, 孔令琳, 赵振华. gga-miR-30a-5p的表达规律及其对鸡脂肪沉积的调控[J]. 中国农业科学, 2025, 58(15): 3134-3144.

HUANG HuaYun, LIU Xing, WANG QianBao, LI RuiRui, YANG MiaoMiao, LI ChunMiao, WU ZhaoLin, KONG LingLin, ZHAO ZhenHua. Expression Pattern of gga-miR-30a-5p and Its Regulation of Abdominal Fat and Intramuscular Fat Deposition in Chicken[J]. Scientia Agricultura Sinica, 2025, 58(15): 3134-3144.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2025.15.014

https://www.chinaagrisci.com/CN/Y2025/V58/I15/3134

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参考文献 32

[1]	陈鑫, 李国强, 孟美瑶, 徐凌燕. 燃烧我的卡路里: 产热脂肪的前世今生. 自然杂志, 2019, 41(6): 423-430.
	CHEN X, LI G Q, MENG M Y, XU L Y. Burning caloric: The stories of thermogenic fat. Chinese Journal of Nature, 2019, 41(6): 423-430. (in Chinese)
[2]	马青山, 王长法, 刘桂芹, 李艳. miRNA调节乳腺组织脂肪代谢的研究进展. 中国畜牧杂志, 2021, 57(5): 40-47.
	MA Q S, WANG C F, LIU G Q, LI Y. Research progress on the role of microRNA in regulation of mammary gland fat metabolism. Chinese Journal of Animal Science, 2021, 57(5): 40-47. (in Chinese)
[3]	田慧月, 王志秀, 周婷婷, 梁文双, 严丹, 董丙强, 常国斌, 陈国宏. 非编码RNAs在家禽脂肪沉积中的作用研究进展. 中国家禽, 2022, 44(3): 85-89.
	TIAN H Y, WANG Z X, ZHOU T T, LIANG W S, YAN D, DONG B Q, CHANG G B, CHEN G H. Research progress on the role of non-coding RNAs in poultry fat deposition. China Poultry, 2022, 44(3): 85-89. (in Chinese)
[4]	CHEN F F, XIONG Y, PENG Y, GAO Y, QIN J, CHU G Y, PANG W J, YANG G S. miR-425-5p inhibits differentiation and proliferation in porcine intramuscular preadipocytes. International Journal of Molecular Sciences, 2017, 18(10): 2101.
[5]	李博. miR-27a靶向CPT1B基因调控绵羊肌间脂肪沉积的研究[D]. 杨凌: 西北农林科技大学, 2022.
	LI B. Regulation of intermuscular fat deposition in sheep by miR-27a targeting CPT1B gene[D]. Yangling: Northwest A & F University, 2022. (in Chinese)
[6]	XU H Y, SHAO J, YIN B Z, ZHANG L M, FANG J C, ZHANG J S, XIA G J. Bovine bta-microRNA-1271 promotes preadipocyte differentiation by targeting activation transcription factor 3. Biochemistry (Moscow), 2020, 85(7): 749-757.
[7]	张珂. 鸡肌肉发育与肌内脂肪沉积关键miRNA鉴定及调控机制解析[D]. 郑州: 河南农业大学, 2024.
	ZHANG K. Identification and regulatory mechanism of key miRNAs in chicken muscle development and intramuscular fat deposition[D]. Zhengzhou: Henan Agricultural University, 2024. (in Chinese)
[8]	池润清, 韩海银, 王鹏, 李凯扬, 储明星, 刘玉芳. 雌激素介导circZNF423作为ceRNA调控oar-miR-541-3p/CALM3通路促进绵羊成肌细胞增殖. 中国农业科学, 2024, 57(3): 597-612. doi: 10.3864/j.issn.0578-1752.2024.03.013.
	CHI R Q, HAN H Y, WANG P, LI K Y, CHU M X, LIU Y F. Estrogen mediates CircZNF423 as a sponge for oar-miR-541-3p to target CALM3 for regulating myoblast proliferation in sheep. Scientia Agricultura Sinica, 2024, 57(3): 597-612. doi: 10.3864/j.issn.0578-1752.2024.03.013. (in Chinese)
[9]	王鹏, 刘子嶷, 刘玉芳, 储明星. mi R-535靶向GAB2基因通过激活PI3K/AKT信号通路促进山羊颗粒细胞增殖. 中国农业科学, 2023, 56(23): 4757-4771. doi: 10.3864/j.issn.0578-1752.2023.23.016.
	WANG P, LIU Z Y, LIU Y F, CHU M X. Mi R-535 targets the GAB2 gene to promote goat granulosa cell proliferation through activation of the PI3K/AKT signaling pathway. Scientia Agricultura Sinica, 2023, 56(23): 4757-4771. doi: 10.3864/j.issn.0578-1752.2023.23.016. (in Chinese)
[10]	JEONG B C, KANG I H, KOH J T. microRNA-302a inhibits adipogenesis by suppressing peroxisome proliferator-activated receptor γ expression. FEBS Letters, 2014, 588(18): 3427-3434.
[11]	KIM S Y, KIM A Y, LEE H W, SON Y H, LEE G Y, LEE J W, LEE Y S, KIM J B. miR-27a is a negative regulator of adipocyte differentiation via suppressing PPARgamma expression. Biochemical and Biophysical Research Communications, 2010, 392(3): 323-328.
[12]	WANG J, GUAN X, GUO F, ZHOU J, CHANG A, SUN B, CAI Y, MA Z, DAI C, LI X, WANG B. miR-30e reciprocally regulates the differentiation of adipocytes and osteoblasts by directly targeting low-density lipoprotein receptor-related protein 6. Cell Death & Disease, 2013, 4(10): e845.
[13]	HU F, WANG M, XIAO T, YIN B Q, HE L Y, MENG W, DONG M J, LIU F. miR-30 promotes thermogenesis and the development of beige fat by targeting RIP140. Diabetes, 2015, 64(6): 2056-2068. doi: 10.2337/db14-1117 pmid: 25576051
[14]	SAHA P K, HAMILTON M P, RAJAPAKSHE K, PUTLURI V, FELIX J B, MASSCHELIN P, COX A R, BAJAJ M, PUTLURI N, COARFA C, HARTIG S M. miR-30a targets gene networks that promote browning of human and mouse adipocytes. American Journal of Physiology Endocrinology and Metabolism, 2020, 319(4): e667-e677.
[15]	HE H R, LI D M, TIAN Y T, WEI Q Y, AMEVOR F K, SUN C J, YU C L, YANG C W, DU H R, JIANG X S, et al. miRNA sequencing analysis of healthy and atretic follicles of chickens revealed that miR-30a-5p inhibits granulosa cell death via targeting Beclin1. Journal of Animal Science and Biotechnology, 2022, 13(1): 55.
[16]	黄华云, 李瑞瑞, 张子晓, 王钱保, 黄正洋, 李春苗, 赵振华. gga-miR-30d-5p表达分析及其对鸡脂肪沉积的调控作用. 南方农业学报, 2023, 54(11): 3359-3368.
	HUANG H Y, LI R R, ZHANG Z X, WANG Q B, HUANG Z Y, LI C M, ZHAO Z H. Expression analysis of gga-miR-30d-5p and its regulation of fat deposition in chickens. Journal of Southern Agriculture, 2023, 54(11): 3359-3368. (in Chinese)
[17]	OUYANG J H, XIE L, NIE Q H, ZENG H, PENG Z J, ZHANG D X, ZHANG X Q. The effects of different sex-linked dwarf variations on Chinese native chickens. Journal of Integrative Agriculture, 2012, 11(9): 1500-1508.
[18]	ZHENG J X, LIU Z Z, YANG N. Deficiency of growth hormone receptor does not affect male reproduction in dwarf chickens. Poultry Science, 2007, 86(1): 112-117. pmid: 17179424
[19]	朱星浩, 陈青, 邵冰豪, 郭钰君, 张向丽, 杜鹏飞, 朱瑶, 黄艳群, 陈文. 杂合型伴性矮小基因对正常体型鸡脂肪沉积的影响. 中国农业科学, 2021, 54(1): 213-223. doi: 10.3864/j.issn.0578-1752.2021. 01.016.
	ZHU X H, CHEN Q, SHAO B H, GUO Y J, ZHANG X L, DU P F, ZHU Y, HUANG Y Q, CHEN W. Effect of the heterozygous sex- linked dwarf gene on fat deposition in normal type chickens. Scientia Agricultura Sinica, 2021, 54(1): 213-223. doi: 10.3864/j.issn.0578-1752.2021.01.016. (in Chinese)
[20]	YU T Y, TIAN X K, LI D, HE Y L, YANG P Y, CHENG Y, ZHAO X, SUN J C, YANG G S. Transcriptome, proteome and metabolome analysis provide insights on fat deposition and meat quality in pig. Food Research International, 2023, 166: 112550.
[21]	YU H W, YU S C, GUO J T, WANG J F, MEI C G, ABBAS RAZA S H, CHENG G, ZAN L S. Comprehensive analysis of transcriptome and metabolome reveals regulatory mechanism of intramuscular fat content in beef cattle. Journal of Agricultural and Food Chemistry, 2024, 72(6): 2911-2924. doi: 10.1021/acs.jafc.3c07844 pmid: 38303491
[22]	苏健, 苏恒. 脂肪细胞分化过程中MAPK信号通路的调控机制. 生命的化学, 2016, 36(2): 252-256.
	SU J, SU H. The regulation mechanism of MAPK signal pathway in adipocyte differentiation. Chemistry of Life, 2016, 36(2): 252-256. (in Chinese)
[23]	ZHAO C Z, WEN Z W, GAO Y F, XIAO F, YAN J Z, WANG X T, MENG T T. Pantothenic acid alleviates fat deposition and inflammation by suppressing the JNK/P38 MAPK signaling pathway. Journal of Medicinal Food, 2024, 27(9): 834-843. doi: 10.1089/jmf.2023.k.0292 pmid: 38949913
[24]	HE X D, SHI J Y, BU L N, ZHOU S T, WU K X, LIANG G, XU X T, WANG A Z. Ursodeoxycholic acid alleviates fat embolism syndrome- induced acute lung injury by inhibiting the p38 MAPK/NF-κB signalling pathway through FXR. Biochemical Pharmacology, 2024, 230(Pt 1): 116574.
[25]	RYU M, KIM M, JUNG H Y, KIM C H, JO C. Effect of p38 inhibitor on the proliferation of chicken muscle stem cells and differentiation into muscle and fat. Animal Bioscience, 2023, 36(2): 295-306.
[26]	SUN X, LIU X Y, WANG C H, REN Z Z, YANG X J, LIU Y L. Deciphering mechanisms of adipocyte differentiation in abdominal fat of broilers. Journal of Agricultural and Food Chemistry, 2024, 72(45): 25403-25413. doi: 10.1021/acs.jafc.4c06867 pmid: 39483088
[27]	COX A R, CHERNIS N, KIM K H, MASSCHELIN P M, SAHA P K, BRILEY S M, SHARP R, LI X, FELIX J B, SUN Z, MOORE D D, PANGAS S A, HARTIG S M. Ube2i deletion in adipocytes causes lipoatrophy in mice. Molecular Metabolism, 2021, 48: 101221.
[28]	SUPAKANKUL P, MEKCHAY S. Effect of UBE3C polymorphisms on intramuscular fat content and fatty acid composition in Duroc pigs. Genetics and Molecular Research, 2016, 15(3): gmr8415.
[29]	WANG Q, LI H X, TAJIMA K, VERKERKE A R P, TAXIN Z H, HOU Z S, COLE J B, LI F, WONG J, ABE I, PRADHAN R N, YAMAMURO T, YONESHIRO T, HIRSCHHORN J N, KAJIMURA S. Post-translational control of beige fat biogenesis by PRDM16 stabilization. Nature, 2022, 609(7925): 151-158.
[30]	LOPEZ-YUS M, FRENDO-CUMBO S, DEL MORAL-BERGOS R, GARCIA-SOBREVIELA M P, BERNAL-MONTERDE V, RYDÉN M, LORENTE-CEBRIAN S, ARBONES-MAINAR J M. CRISPR/ Cas9-mediated deletion of adipocyte genes associated with NAFLD alters adipocyte lipid handling and reduces steatosis in hepatocytes in vitro. American Journal of Physiology Cell Physiology, 2023, 325(5): c1178-c1189.
[31]	WANG Y J, LIU W T, LIU Y D, CUI J L, ZHAO Z W, CAO H, FU Z, LIU B. Long noncoding RNA H19 mediates LCoR to impact the osteogenic and adipogenic differentiation of mBMSCs in mice through sponging miR-188. Journal of Cellular Physiology, 2018, 233(9): 7435-7446. doi: 10.1002/jcp.26589 pmid: 29663375
[32]	XIAO F, TANG C Y, TANG H N, WU H X, HU N, LI L, ZHOU H D. Long non-coding RNA 332443 inhibits preadipocyte differentiation by targeting Runx1 and p38-MAPK and ERK1/2-MAPK signaling pathways. Frontiers in Cell and Developmental Biology, 2021, 9: 663959.

通路 Pathway	P值P value	富集基因Enriched gene
MAPK信号通路 MAPK signaling pathway	0.0001	BDNF, NFATC3, CACNA1D, CACNA1C, CRKL, RAP1B, CACNB2, PPM1A, PPP3CA, MAPK8, PPP3CB, PPP3R1, GDNF, TAOK1, RASA1, RASA2, NF1, RAPGEF2, KRAS, SOS1, MAP4K4, MAP3K5
泛素介导的蛋白水解 Ubiquitin mediated proteolysis	0.0012	UBE2F, UBE2I, UBE3C, CUL2, UBE2D3, NEDD4L, WWP1, CBLB, UBE2J1, SOCS3, HERC2, NEDD4, TRIP12
多聚酶抑制复合体 Polycomb repressive complex	0.0025	CBX8, PCGF5, EED, ASXL3, PCGF3, UBE2D3, LCOR, BCOR, YAF2
紧密连接 Tight junction	0.0337	RAP2C, MAPK8, NEDD4, ACTN1, NEDD4L, RAPGEF2, CACNA1D, AMOTL2, RAB8A, MAP3K5, RUNX1