白藜芦醇通过PROX1/SIRT1信号通路调控山羊肌纤维类型转化的机制研究

doi:10.11843/j.issn.0366-6964.2025.01.019

Abstract

Abstract:

The aim of this study was to analyze the molecular mechanism of resveratrol (RES) regulating myofiber type transformation by mediating PROX1 gene in goat skeletal muscle. In vivo component, twelve 180-day-old depopulated Nubian goats ((28.25±0.26) kg) were randomly divided into two groups, with 6 replicates each, the PROX1 gene expression levels of in dorsal longest muscle tissues supplemented with 0 and 150 mg·kg^-1 RES were detected by real-time fluorescence quantitative PCR (RT-qPCR) and Western blot. In vitro test, based on the previous research results, 20 μmol·L^-1 RES was identified as the optimal concentration for cell culture. PROX1 interference fragments and a combination of interference fragments with 20 μmol·L^-1 RES were transfected into goat skeletal muscle myoblasts. The expression levels of key genes in the CaN/NFAT pathway and the marker genes for the four muscle fiber types were measured at both mRNA and protein levels. Meanwhile, protein-protein docking technology was used to detect whether SIRT1 interacts with the PROX1 protein interact, and the regulatory relationship between SIRT1 and the CaN/NFAT pathway was verified by co-treatment of goat myoblasts with the SIRT1 inhibitor EX-527 and RES. The results showed that, compared with the control group, the expression of the SIRT1 gene and key genes in the CaN/NFAT pathway at the mRNA level in goat myoblasts treated with 150 mg·kg^-1 RES was significantly upregulated (P < 0.001), the expression of MYH7 protein was significantly upregulated (P < 0.01), and the expression of MYH4 protein was significantly downregulated (P < 0.01). After PROX1 gene interference, the mRNA and protein expression levels of key genes in the CaN/NFAT pathway, MYH7 and MYH4 showed opposite trends, which were altered when the interference fragments and RES co-treated the myoblasts. The protein-protein docking results indicated that SIRT1 and PROX1 proteins could interact. After myoblasts treated with the SIRT1 inhibitor, the mRNA expression levels of key genes (NFATc1, TNNC1, TNNI1, MYOZ2, SLN) in the CaN/NFAT pathway were significantly downregulated compared to the control group (P < 0.01). Following RES intervention, the mRNA expression levels of TNNC1 and TNNI1 were significantly upregulated (P < 0.01). This study initially revealed the mechanism of RES regulating the transformation of muscle fiber types through SIRT1/PROX1/CaN/NFAT pathway, which provides a theoretical basis for the subsequent improvement of livestock and poultry meat quality.

Key words: resveratrol(RES), CaN/NFAT signaling pathway, goat myoblasts, myofiber transformation

CLC Number:

S827.1

WU Shaoqiang, LIU Yufan, WEI Yirong, HUANG Yanna, JIANG Qinyang. Mechanism of Resveratrol Regulating Myofiber Type Transformation through PROX1/SIRT1 Signaling Pathway in Goats[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 201-212.

Figures/Tables 9

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References 35

1	HOU Y R , LIU C , SU L , et al. Dietary linseed supplementation improves meat quality and flavor of sheep by altering muscle fiber characteristics and antioxidant capacity[J]. Anim Sci J, 2023, 94 (1): e13801. doi: 10.1111/asj.13801
2	SAWANO S , MIZUNOYA W . History and development of staining methods for skeletal muscle fiber types[J]. Histol Histopathol, 2022, 37 (6): 493- 503.
3	BOTTINELLI R , REGGIANI C . Human skeletal muscle fibres: molecular and functional diversity[J]. Prog Biophys Mol Biol, 2000, 73 (2-4): 195- 262. doi: 10.1016/S0079-6107(00)00006-7
4	谢兵红, 单艳菊, 范晨宇, 等. 家禽骨骼肌肌纤维类型影响因素及其转化的研究进展[J]. 中国畜牧兽医, 2024, 51 (4): 1561- 1572.
	XIE B H , SHAN Y J , FAN C Y , et al. Research progress on influencing factors and transformation of skeletal muscle fiber types in poultry[J]. China Animal Husbandry & Veterinary Medicine, 2024, 51 (4): 1561- 1572.
5	WENG K Q , HUO W R , LI Y , et al. Fiber characteristics and meat quality of different muscular tissues from slow-and fastgrowing broilers[J]. Poult Sci, 2022, 101 (1): 101537. doi: 10.1016/j.psj.2021.101537
6	MEUNIER B , PICARD B , ASTRUC T , et al. Development of image analysis tool for the classification of muscle fibre type using immunohistochemical staining[J]. Histochem Cell Biol, 2010, 134 (3): 307- 317. doi: 10.1007/s00418-010-0733-7
7	袁继红, 龙欢, 田明芳, 等. 猪Prox1基因的染色体定位和亚细胞定位的研究[J]. 畜牧兽医学报, 2013, 44 (3): 351- 357.
	YUAN J H , LONG H , TIAN M F , et al. Study on chromosomal and subcellular localization for porcine Prox1 gene[J]. Acta Veterinaria et Zootechnica Sinica, 2013, 44 (3): 351- 357.
8	KIVELÄ R , SALMELA I , NGUYEN Y H , et al. The transcription factor Prox1 is essential for satellite cell differentiation and muscle fibre-type regulation[J]. Nat Commun, 2016, 7, 13124. doi: 10.1038/ncomms13124
9	CHEN X L , ZHANG M , JIA G , et al. L-theanine induces skeletal muscle fiber type transformation by activation of prox1/CaN signaling pathway in C2C12 myotubes[J]. Biol Chem, 2022, 403 (10): 959- 967. doi: 10.1515/hsz-2022-0165
10	PETCHEY L K , RISEBRO C A , VIEIRA J M , et al. Loss of Prox1 in striated muscle causes slow to fast skeletal muscle fiber conversion and dilated cardiomyopathy[J]. Proc Natl Acad Sci U S A, 2014, 111 (26): 9515- 9520. doi: 10.1073/pnas.1406191111
11	WU H , ROTHERMEL B , KANATOUS S , et al. Activation of MEF2 by muscle activity is mediated through a calcineurin-dependent pathway[J]. EMBO J, 2001, 20 (22): 6414- 6423. doi: 10.1093/emboj/20.22.6414
12	田佳敏. 穴位埋线对运动性疲劳模型大鼠骨骼肌的影响及CaN/NFAT信号通路的研究[D]. 呼和浩特: 内蒙古农业大学, 2023.
	TIAN J M. Effects of acupuncture point embedding on skeletal muscle of rats with exercise fatigue model and study of CaN/NFAT signaling pathway[D]. Hohhot: Inner Mongolia Agricultural University, 2023. (in Chinese)
13	单艳菊, 宋迟, 束婧婷, 等. 钙调磷酸酶催化亚基A基因(PPP3CA)在不同品种鸭发育早期肌肉中的表达及其与肌纤维特性的相关性[J]. 农业生物技术学报, 2015, 23 (2): 236- 243. doi: 10.3969/j.issn.1674-7968.2015.02.011
	SHAN Y J , SONG C , SHU J T , et al. Expression profile of protein phosphatase 3 catalytic A gene(PPP3CA) mRNA in muscles of two duck breeds (Anas platyrhynchos domestica) and its relationship with myofiber traits during early development[J]. Journal of Agricultural Biotechnology, 2015, 23 (2): 236- 243. doi: 10.3969/j.issn.1674-7968.2015.02.011
14	PAN S , KOYANO-NAKAGAWA N , TSURUTA L , et al. Molecular cloning and functional characterization of murine cDNA encoding transcription factor NFATc[J]. Biochem Biophys Res Commun, 1997, 240 (2): 314- 323. doi: 10.1006/bbrc.1997.7381
15	RAO A , LUO C , HOGAN P G . Transcription factors of the NFAT family: regulation and function[J]. Annu Rev Immunol, 1997, 15, 707- 747. doi: 10.1146/annurev.immunol.15.1.707
16	SCHULZ R A , YUTZEY K E . Calcineurin signaling and NFAT activation in cardiovascular and skeletal muscle development[J]. Dev Biol, 2004, 266 (1): 1- 16. doi: 10.1016/j.ydbio.2003.10.008
17	DU J J , SHEN L Y , ZHANG P W , et al. The regulation of skeletal muscle fiber-type composition by betaine is associated with NFATc1/MyoD[J]. J Mol Med (Berl), 2018, 96 (7): 685- 700. doi: 10.1007/s00109-018-1657-2
18	HASAN M , BAE H . An overview of stress-induced resveratrol synthesis in grapes: perspectives for resveratrol-enriched grape products[J]. Molecules, 2017, 22 (2): 294. doi: 10.3390/molecules22020294
19	REPOSSI G , DAS U N , EYNARD A R . Molecular basis of the beneficial actions of resveratrol[J]. Arch Med Res, 2020, 51 (2): 105- 114. doi: 10.1016/j.arcmed.2020.01.010
20	ZHANG S B , PAN P , XIE H Y , et al. Resveratrol improves meat quality traits by activating the lncRNAs-KEAP1-NRF2 axis in pigs[J]. Meat Sci, 2024, 209, 109411. doi: 10.1016/j.meatsci.2023.109411
21	SHEN Y J , JIANG Y H , ZHANG S B , et al. The effect of dietary supplementation with resveratrol on growth performance, carcass and meat quality, blood lipid levels and ruminal microbiota in fattening goats[J]. Foods, 2022, 11 (4): 598. doi: 10.3390/foods11040598
22	HUANG B Q , ZHANG W G , WEI L T , et al. Resveratrol down-regulates endothelin type B receptors in vascular smooth muscle cells via Sirt1/ERK1/2/NF-кB signaling pathways[J]. Eur J Pharmacol, 2018, 840, 44- 49. doi: 10.1016/j.ejphar.2018.09.022
23	张静月, 董鹏程, 左惠心, 等. 白藜芦醇通过SIRT1/PGC-1α影响牛肌管细胞线粒体生物发生和肌纤维类型转化[J]. 食品科学, 2024, 45 (4): 1- 9.
	ZHANG J Y , DONG P C , ZUO H X , et al. Effect of resveratrol on mitochondrial biogenesis and muscle fiber type transformation in bovine myotubes via SIRT1/PGC-1α[J]. Food Science, 2024, 45 (4): 1- 9.
24	刘雨帆, 邹菊红, 何海恩, 等. 努比亚山羊(Capra nubiana)PROX1基因克隆及功能研究[J]. 基因组学与应用生物学, 2024, 43 (7): 1214- 1224.
	LIU Y F , ZOU J H , HE H E , et al. Cloning and functional study of PROX1 gene in Nubian goat (Capra nubiana)[J]. Genomics and Applied Biology, 2024, 43 (7): 1214- 1224.
25	庾蕾, 刘建平, 庄志雄, 等. 实时RT-PCR基因表达相对定量REST^ⓒ软件分析与2^(-ΔΔCT)法比较[J]. 热带医学杂志, 2007, 7 (10): 956- 958. doi: 10.3969/j.issn.1672-3619.2007.10.006
	YU L , LIU J P , ZHUANG Z X , et al. Quantitative analysis of real-time PCR expression production by REST^ⓒ and 2^(-ΔΔCT)[J]. Journal of Tropical Medicine, 2007, 7 (10): 956- 958. doi: 10.3969/j.issn.1672-3619.2007.10.006
26	张宁, 康燕, 任文义, 等. 饲粮添加琥珀酸对滩羊肉品质及肌纤维类型的影响[J]. 动物营养学报, 2023, 35 (10): 6538- 6545. doi: 10.12418/CJAN2023.598
	ZHANG N , KANG Y , REN W Y , et al. Effects of dietary succinic acid on meat quality and muscle fiber type of Tan Sheep[J]. Journal of Animal Nutrition, 2023, 35 (10): 6538- 6545. doi: 10.12418/CJAN2023.598
27	LJUBICIC V , BURT M , LUNDE J A , et al. Resveratrol induces expression of the slow, oxidative phenotype in mdx mouse muscle together with enhanced activity of the SIRT1-PGC-1α axis[J]. Am J Physiol Cell Physiol, 2014, 307 (1): C66- C82. doi: 10.1152/ajpcell.00357.2013
28	孙静, 左惠心, 刘昀阁, 等. 白藜芦醇通过Akt/FoxO信号通路促进牛肌纤维类型转化的研究[J/OL]. 食品与发酵工业, doi: 10.13995/j.cnki.11-1802/ts.038911.
	SUN J, ZUO H X, LIU Y G, et al. The study of resveratrol promotes the transformation of bovine muscle fiber types through Akt/FoxO signaling pathway[J/OL]. Food and Fermentation Industry, doi: 10.13995/j.cnki.11-1802/ts.038911.(in Chinese)
29	张彩云, 康相涛. 白藜芦醇对肉仔鸡抗氧化能力和肉品质的影响[J]. 江苏农业学报, 2011, 27 (3): 587- 591. doi: 10.3969/j.issn.1000-4440.2011.03.022
	ZHANG C Y , KANG X T . Effects of dietary resveratrol on antioxidant ability and meat quality in broilers[J]. Jiangsu Journal of Agricultural Sciences, 2011, 27 (3): 587- 591. doi: 10.3969/j.issn.1000-4440.2011.03.022
30	ZHANG C , LUO J Q , YU B , et al. Dietary resveratrol supplementation improves meat quality of finishing pigs through changing muscle fiber characteristics and antioxidative status[J]. Meat Sci, 2015, 102, 15- 21. doi: 10.1016/j.meatsci.2014.11.014
31	CAO D F , WANG M Z , QIU X Y , et al. Structural basis for allosteric, substrate-dependent stimulation of SIRT1 activity by resveratrol[J]. Genes Dev, 2015, 29 (12): 1316- 1325. doi: 10.1101/gad.265462.115
32	ZHANG J Y , LI J Q , LIU Y G , et al. Effect of resveratrol on skeletal slow-twitch muscle fiber expression via AMPK/PGC-1α signaling pathway in bovine myotubes[J]. Meat Sci, 2023, 204, 109287. doi: 10.1016/j.meatsci.2023.109287
33	WANG R T , YUAN W D , LI L , et al. Resveratrol ameliorates muscle atrophy in chronic kidney disease via the axis of SIRT1/FoxO1[J]. Phytother Res, 2022, 36 (8): 3265- 3275. doi: 10.1002/ptr.7499
34	黄静. SIRT1抑制剂EX-527通过抑制mTOR的激活减轻内毒素诱导的急性肺损伤[D]. 重庆: 重庆医科大学, 2018.
	HUANG J. The SIRT1 inhibitor EX-527 suppresses mTOR activation and alleviates acute lung injury in mice with endotoxiemia[D]. Chongqing: Chongqing Medical University, 2018. (in Chinese)
35	杨冠青, 刘娟, 杨阳, 等. 猪TNNC1基因和TNNC2基因在肌肉组织中的发育性表达研究[J]. 国外畜牧学(猪与禽), 2020, 40 (5): 21- 26.
	YANG G Q , LIU J , YANG Y , et al. Developmental expression of porcine TNNC1 and TNNC2 genes in muscle tissue[J]. Animal Science Abroad (Pigs and Poultry), 2020, 40 (5): 21- 26.

基因 Gene	登录号 Accession number	引物序列(5′→3′) Primer sequence	产物长度/bp Amplification length
MYH7	XM_018054604	F: AGTTCGTCAAGGCCACGATT R: CCTCCTTCACGGTCACTGTC	87
MYH2	XM_018064659	F: GATGGTGGAGAGGAGGGAGT R: TCAGCAGGGGCTTGATCTTG	113
MYH4	XM_018064656	F: GCACCAGCGCCTAATCAATG R: TCGGGATAGCTGAGACACCA	115
MYH1	XM_018064657	F: AATCCTGGGCTTCACTTCGG R: CTGGTTCTGCTTGCTCCTCA	116
SIRT1	NM_001314319	F: ATTTTCCATGGCGCTGAGGT R: GGACTCTGGCATGTTCCACT	98
NFATc1	XM_018039660	F: GTCTGCAACGGGAAGAGGAA R: TCGTAATCATCGCTGGGCTC	98
TNNC1	NM_001285572	F: TGACATCTACAAGGCTGCGG R: CTGCCATCCTCATCCACCTC	201
TNNI1	XM_018060884	F: GCCAACCTCAAGTCCGTGAA R: CCTCCACGTTCTTCCTCCAG	82
MYOZ2	NM_001287568	F: GTCATGCACCTGGGCAAAAA R: GGCACCACGGTTACTGAGAT	81
SLN	XM_005689416	F: GATCCACCCGGGAGATCTGT R: ATAGGACCTCACAAGGAGCCA	80
PROX1	XM_013970386	F: CTCTCTAGTACAGGCTCCGAAG R: GAATGGCTCATACCCCGGAT	191
GAPDH	XM_005680968	F: CCCGTTCGACAGATAGCCGTA R: AATCCGTTCACTCCGACCT	83

Mechanism of Resveratrol Regulating Myofiber Type Transformation through PROX1/SIRT1 Signaling Pathway in Goats

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