β-羟基丁酸通过TRIB3/Akt/mTOR通路调控蛋白质合成和降解平衡改善卒中相关性肌少症

doi:10.3969/j.issn.1673-5765.2025.09.012

中国卒中杂志 ›› 2025, Vol. 20 ›› Issue (9): 1167-1178.DOI: 10.3969/j.issn.1673-5765.2025.09.012

β-羟基丁酸通过TRIB3/Akt/mTOR通路调控蛋白质合成和降解平衡改善卒中相关性肌少症

黄媛媛^1*，周婉冰^2*，李小燕³，卢政红⁴，洪小丹⁵（*第一作者）

1 东莞 523808 东莞职业技术学院卫生健康学院
2 南方医科大学基础医学院
3 赣州市人民医院消化科内镜中心
4 南方医科大学南方医院消化科胃镜中心
5 南方医科大学南方医院神经内科

收稿日期:2025-01-10 修回日期:2025-08-05 接受日期:2025-08-12 出版日期:2025-09-20 发布日期:2025-09-20
通讯作者: 洪小丹 936461011@qq.com

β-Hydroxybutyrate Improves Stroke-Related Sarcopenia by Regulating Protein Synthesis and Degradation Homeostasis via the TRIB3/Akt/mTOR Pathway

HUANG Yuanyuan^1*, ZHOU Wanbing^2*, LI Xiaoyan³, LU Zhenghong⁴, HONG Xiaodan⁵ (*contributed equally)

1 School of Health, Dongguan Polytechnic, Dongguan 523808, China
2 School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China
3 Endoscopy Center of Gastroenterology Department, Ganzhou People’s Hospital, Ganzhou 341199, China
4 Gastroscopy Center of Gastroenterology Department, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China
5 Department of Neurology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China

Received:2025-01-10 Revised:2025-08-05 Accepted:2025-08-12 Online:2025-09-20 Published:2025-09-20
Contact: HONG Xiaodan, E-mail: 936461011@qq.com

摘要/Abstract

摘要： 目的探讨β-羟基丁酸（β-hydroxybutyrate，BHB）是否经Tribbles同源蛋白3（Tribbles homolog 3，TRIB3）/蛋白激酶B（protein kinase B，Akt）/哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）通路调控蛋白质合成和降解平衡，进而改善卒中相关性肌少症（stroke-related sarcopenia，SRS）。
方法采用8周龄雄性C57BL/6J小鼠，通过线栓法建立短暂性大脑中动脉栓塞（transient middle cerebral artery occlusion，tMCAO）模型。将造模后小鼠随机分为BHB组与对照组：BHB组皮下注射5 mg/kg BHB，每8小时1次，持续3 d；对照组注射等体积生理盐水。测量小鼠脑梗死体积；采用转棒实验、抓握力强度测试和悬挂实验评估小鼠骨骼肌力量；取小鼠双侧胫骨前肌组织样本进行苏木精-伊红染色，比较肌纤维横截面积。通过生物信息学分析筛选tMCAO小鼠与假手术组小鼠相比，胫骨前肌组织中的差异基因富集通路；采用免疫印迹法、实时定量PCR检测Akt/mTOR通路及蛋白质合成和降解相关基因在不同组中的表达；采用STRING数据库预测Akt相互作用蛋白，并通过免疫共沉淀验证TRIB3与Akt蛋白-蛋白相互作用。
结果与对照组相比，造模后BHB组小鼠的脑梗死体积下降（P<0.001）；转棒时间、握力及悬挂时间增加（P<0.001）；肌纤维横截面积增大（P<0.001）。差异表达基因在磷脂酰肌醇3激酶（phosphoinositide 3-kinase，PI3K）/Akt通路上富集最为显著。BHB组磷酸化哺乳动物雷帕霉素靶蛋白（phosphorylated mTOR，p-mTOR）及磷酸化蛋白激酶B（phosphorylated Akt，p-Akt）蛋白表达增加（P<0.001）；蛋白质降解调控基因叉头盒蛋白O3a（forkhead box protein O3a，FOXO3a）、蛋白质降解相关基因肌肉环指蛋白1（muscle ring-finger protein-1，MuRF1）和肌肉萎缩F盒（muscle atrophy F-box，MAFbx）表达水平降低（P<0.001），而蛋白质合成相关基因p70核糖体蛋白S6激酶（p70 ribosomal protein S6 kinase，p70S6K）表达水平升高（P<0.001）。免疫共沉淀证明TRIB3与Akt存在相互作用，且BHB处理后TRIB3表达水平下降（P<0.001）。
结论 BHB可通过抑制TRIB3表达，促进Akt磷酸化，从而激活Akt/mTOR通路，促进蛋白质合成并抑制降解，改善SRS。

文章导读： 本研究揭示了β-羟基丁酸不仅能减少小鼠卒中相关性肌少症模型的脑梗死体积、改善神经功能，还可增强骨骼肌蛋白质合成、抑制其降解，缓解卒中后骨骼肌萎缩。本研究结果还提示，β-羟基丁酸对卒中相关性肌少症的作用机制可能与调控TRIB3/Akt/mTOR通路相关。

Abstract: Objective To investigate whether β-hydroxybutyrate (BHB) alleviates stroke-related sarcopenia (SRS) by regulating protein synthesis and degradation homeostasis via the Tribbles homolog 3 (TRIB3) / protein kinase B (Akt) / mammalian target of rapamycin (mTOR) pathway.
Methods Eight-week-old male C57BL/6J mice were used to establish the transient middle cerebral artery occlusion (tMCAO) model by the suture method. After modeling, the mice were randomly divided into the BHB group and the control group: the BHB group was subcutaneously injected with BHB at a dose of 5 mg/kg, once every 8 hours for 3 consecutive days; the control group was injected with an equal volume of physiological saline. The cerebral infarction volume was measured. The skeletal muscle strength of mice was evaluated by the rotarod test, grip strength test, and hanging test. Bilateral tibialis anterior muscle tissue samples of mice were collected for hematoxylin and eosin staining to compare the cross-sectional area of muscle fibers. Bioinformatics analysis was used to screen the differential gene enrichment pathways in the tibialis anterior muscle tissue of tMCAO mice compared with the shame-operation group. Western blot and Real time quantitative PCR were used to detect the expression of Akt/mTOR pathway-related molecules and protein synthesis and degradation-related genes in different groups. The STRING database was used to predict Akt-interacting proteins, and co-immunoprecipitation was performed to verify the protein-protein interaction between TRIB3 and Akt.
Results After modeling, the cerebral infarction volume in mice of the BHB group was reduced compared with the control group (P<0.001), the rotarod duration, grip strength, and hanging time were increased (P<0.001) and the cross-sectional area of muscle fibers (P<0.001) were increased. The phosphoinositide 3-kinase (PI3K) / Akt pathway showed the most significant enrichment of differentially expressed genes. The protein expressions of phosphorylated mTOR (p-mTOR) and phosphorylated Akt (p-Akt) were increased in the BHB group (P<0.001). The expression levels of the protein degradation regulatory gene forkhead box protein O3a (FOXO3a) and protein degradation-related genes muscle ring-finger protein-1 (MuRF1) and muscle atrophy F-box (MAFbx) were decreased (P<0.001), while the expression level of the protein synthesis-related gene p70 ribosomal protein S6 kinase (p70S6K) was increased (P<0.001). Co-immunoprecipitation showed that TRIB3 interacted with Akt, and the expression level of TRIB3 decreased after BHB treatment (P<0.001).
Conclusions BHB can promote Akt phosphorylation by inhibiting TRIB3 expression, thereby activating the Akt/mTOR pathway, facilitating protein synthesis and inhibiting degradation, and ultimately improving SRS.

Key words: Stroke-related sarcopenia; β-hydroxybutyrate; Tribbles homolog 3; Middle cerebral artery occlusion

中图分类号:

黄媛媛, 周婉冰, 李小燕, 卢政红, 洪小丹. β-羟基丁酸通过TRIB3/Akt/mTOR通路调控蛋白质合成和降解平衡改善卒中相关性肌少症[J]. 中国卒中杂志, 2025, 20(9): 1167-1178.

HUANG Yuanyuan, ZHOU Wanbing, LI Xiaoyan, LU Zhenghong, HONG Xiaodan. β-Hydroxybutyrate Improves Stroke-Related Sarcopenia by Regulating Protein Synthesis and Degradation Homeostasis via the TRIB3/Akt/mTOR Pathway[J]. Chinese Journal of Stroke, 2025, 20(9): 1167-1178.

参考文献

[1] HUANG C Y，LIU Y H. Sex difference，proteostasis and mitochondrial function impact stroke-related sarcopenia—a systematic review and meta-analysis[J/OL]. Ageing Res Rev，2024，101：102484[2025-08-10]. https://doi.org/10.1016/j.arr.2024.102484.
[2] RYAN A S，DOBROVOLNY C L，SMITH G V，et al. Skeletal muscle hypertrophy and muscle myostatin reduction after resistive training in stroke survivors[J/OL]. Stroke，2011，42（2）：416-420[2025-08-10]. https://doi.org/10.1161/STROKEAHA.110.602441.
[3] CHON J，SOH Y，SHIM G Y. Stroke-related sarcopenia：pathophysiology and diagnostic tools[J/OL]. Brain Neurorehabil，2024，17（3）：e23[2025-08-10]. https://doi.org/10.12786/bn.2024.17.e23.
[4] SU Y，YUKI M，OTSUKI M. Prevalence of stroke-related sarcopenia：a systematic review and meta-analysis[J/OL]. J Stroke Cerebrovasc Dis，2020，29（9）：105092[2025-08-10]. https://doi.org/10.1016/j.jstrokecerebrovasdis.2020.105092.
[5] 骆晓靖，胡填，古剑雄. 脑卒中相关性肌少症的研究进展[J]. 实用心脑肺血管病杂志，2025，33（3）：136-140.
LUO X J，HU T，GU J X. Research progress of stroke-related sarcopenia[J]. Pract J Cardiac Cereb Pneum Vasc Dis，2025，33（3）：136-140.
[6] 陈雪婷，杨雪，任欢欢，等. 脑卒中相关性肌少症的临床评估进展[J]. 中华物理医学与康复杂志，2025，47（1）：91-94.
CHEN X T，YANG X，REN H H，et al. Advances in the clinical evaluation of stroke-associated sarcopenia[J]. Chin J Phys Med Rehabil，2025，47（1）：91-94.
[7] WHITE H，VENKATESH B，JONES M，et al. Inducing ketogenesis via an enteral formulation in patients with acute brain injury：a phase Ⅱ study[J]. Neurol Res，2020，42（4）：275-285.
[8] MILLER V J，VILLAMENA F A，VOLEK J S. Nutritional ketosis and mitohormesis：potential implications for mitochondrial function and human health[J/OL]. J Nutr Metab，2018：5157645[2025-08-10]. https://doi.org/10.1155/2018/5157645.
[9] NEWMAN J C，VERDIN E. β-hydroxybutyrate：a signaling metabolite[J/OL]. Annu Rev Nutr，2017，37：51-76[2025-08-10]. https://doi.org/10.1146/annurev-nutr-071816-064916.
[10] LIN C M，WANG S N，WEI X M，et al. β-hydroxybutyrate inhibits FOXO3a by histone H3K9 β-hydroxybutyrylation to ameliorate stroke-related sarcopenia[J/OL]. J Funct Foods，2024，120：106365[2025-08-10]. https://doi.org/10.1016/j.jff.2024.106365.
[11] BODINE S C，STITT T N，GONZALEZ M，et al. Akt/mTOR pathway is a crucial regulator of skeletal muscle hypertrophy and can prevent muscle atrophy in vivo[J]. Nat Cell Biol，2001，3（11）：1014-1019.
[12] CHOI R H，MCCONAHAY A，SILVESTRE J G，et al. TRB3 regulates skeletal muscle mass in food deprivation-induced atrophy[J]. FASEB J，2019，33（4）：5654-5666.
[13] ZHU J，LI Z Q，JI Z，et al. Glycocalyx is critical for blood-brain barrier integrity by suppressing caveolin1-dependent endothelial transcytosis following ischemic stroke[J/OL]. Brain Pathol，2022，32（1）：e13006[2025-08-10]. https://doi.org/10.1111/bpa.13006.
[14] GOOSSENS C，WECKX R，DERDE S，et al. Adipose tissue protects against sepsis-induced muscle weakness in mice：from lipolysis to ketones[J/OL]. Crit Care，2019，23（1）：236[2025-08-10]. https://doi.org/10.1186/s13054-019-2506-6.
[15] CHIANG T，MESSING R O，CHOU W H. Mouse model of middle cerebral artery occlusion[J/OL]. J Vis Exp，2011（48）：2761[2025-08-10]. https://doi.org/10.3791/2761.
[16] FERRANDI P J，KHAN M M，PAEZ H G，et al. Transcriptome analysis of skeletal muscle reveals altered proteolytic and neuromuscular junction associated gene expressions in a mouse model of cerebral ischemic stroke[J/OL]. Genes（Basel），2020，11（7）：726[2025-08-10]. https://doi.org/10.3390/genes11070726.
[17] YOUM Y H，NGUYEN K Y，GRANT R W，et al. The ketone metabolite β-hydroxybutyrate blocks NLRP3 inflammasome-mediated inflammatory disease[J]. Nat Med，2015，21（3）：263-269.
[18] CAMBEROS-LUNA L，GERÓNIMO-OLVERA C，MONTIEL T，et al. The ketone body，β-hydroxybutyrate stimulates the autophagic flux and prevents neuronal death induced by glucose deprivation in cortical cultured neurons[J]. Neurochem Res，2016，41（3）：600-609.
[19] VANDOORNE T，DE SMET S，RAMAEKERS M，et al. Intake of a ketone ester drink during recovery from exercise promotes mTORC1 signaling but not glycogen resynthesis in human muscle[J/OL]. Front Physiol，2017，8：310[2025-08-10]. https://doi.org/10.3389/fphys.2017.00310.
[20] ZOU X T，MENG J，LI L，et al. Acetoacetate accelerates muscle regeneration and ameliorates muscular dystrophy in mice[J]. J Biol Chem，2016，291（5）：2181-2195.
[21] YOON M S. mTOR as a key regulator in maintaining skeletal muscle mass[J/OL]. Front Physiol，2017，8：788[2025-08-10]. https://doi.org/10.3389/fphys.2017.00788.
[22] OH H J，JIN H，LEE J Y，et al. Silk peptide ameliorates sarcopenia through the regulation of Akt/mTOR/FOXO3a signaling pathways and the inhibition of low-grade chronic inflammation in aged mice[J/OL]. Cells，2023，12（18）：2257[2025-08-10]. https://doi.org/10.3390/cells12182257.
[23] 王辉强，李玉环. Tribbles同源蛋白3的功能研究进展[J]. 药学学报，2018，53（12）：1986-1994.
WANG H Q，LI Y H. Progress in the study of the function of Tribbles homologous protein 3[J]. Acta Pharmaceutica Sinica，2018，53（12）：1986-1994.
[24] KISS-TOTH E，BAGSTAFF S M，SUNG H Y，et al. Human Tribbles，a protein family controlling mitogen-activated protein kinase cascades[J]. J Biol Chem，2004，279（41）：42703-42708.
[25] HUANG J，TENG L，LIU T，et al. Identification of a novel serine/threonine kinase that inhibits TNF-induced NF-κB activation and p53-induced transcription[J]. Biochem Biophys Res Commun，2003，309（4）：774-778.
[26] WU M，XU L G，ZHAI Z H，et al. SINK is a p65-interacting negative regulator of NF-κB-dependent transcription[J]. J Biol Chem，2003，278（29）：27072-27079.
[27] EYERS P A，KEESHAN K，KANNAN N. Tribbles in the 21st century：the evolving roles of Tribbles pseudokinases in biology and disease[J]. Trends Cell Biol，2017，27（4）：284-298.
[28] ZHOU H J，WANG X Y，WANG L Q，et al. Interfering TRIB3 protects the blood brain barrier through PI3K/Akt pathway to alleviate cerebral ischemia-reperfusion injury in diabetes mellitus mice[J/OL]. Chem Biol Interact，2024，387：110807[2025-08-10]. https://doi.org/10.1016/j.cbi.2023.110807.
[29] SALAZAR M，LORENTE M，GARCÍA-TABOADA E，et al. Loss of Tribbles pseudokinase-3 promotes Akt-driven tumorigenesis via FOXO inactivation[J]. Cell Death Differ，2015，22（1）：131-144.
[30] CHOI R H，MCCONAHAY A，JEONG H W，et al. Tribbles 3 regulates protein turnover in mouse skeletal muscle[J]. Biochem Biophys Res Commun，2017，493（3）：1236-1242.
[31] KATO S，DU K Y. TRB3 modulates C2C12 differentiation by interfering with Akt activation[J]. Biochem Biophys Res Commun，2007，353（4）：933-938.
[32] KO C H，WU S J，WANG S T，et al. Effects of enriched branched-chain amino acid supplementation on sarcopenia[J]. Aging（Albany NY），2020，12（14）：15091-15103.
[33] KANG D，PARK J. A community-based resistance training exercise for post-stroke patients with sarcopenia：bridging institutional and community-based rehabilitation in a multicenter，randomized controlled trial[J/OL]. Life（Basel），2025，15（5）：748[2025-08-10]. https://doi.org/10.3390/life15050748.
[34] LAPLANTE M，SABATINI D M. mTOR signaling in growth control and disease[J]. Cell，2012，149（2）：274-293.
[35] YANG H J，SHAN W，ZHU F，et al. Ketone bodies in neurological diseases：focus on neuroprotection and underlying mechanisms[J/OL]. Front Neurol，2019，10：585[2025-08-10]. https://doi.org/10.3389/fneur.2019.00585.
[36] MA X M，BLENIS J. Molecular mechanisms of mTOR-mediated translational control[J]. Nat Rev Mol Cell Biol，2009，10（5）：307-318.
[37] FENG G，WU Z K，YANG L Y，et al. β-hydroxybutyrate and ischemic stroke：roles and mechanisms[J/OL]. Mol Brain，2024，17（1）：48[2025-08-10]. https://doi.org/10.1186/s13041-024-01119-0.
[38] WANG Z T，LI T，DU M Y，et al. β-hydroxybutyrate improves cognitive impairment caused by chronic cerebral hypoperfusion via amelioration of neuroinflammation and blood-brain barrier damage[J/OL]. Brain Res Bull，2023，193：117-130[2025-08-10]. https://doi.org/10.1016/j.brainresbull.2022.12.011.
[39] LIU G M，JIANG S D，XIE W Q，et al. Biomarkers for sarcopenia，muscle mass，muscle strength，and physical performance：an umbrella review[J/OL]. J Transl Med，2025，23（1）：650[2025-08-10]. https://doi.org/10.1186/s12967-025-06575-3.
[40] HOLSTEIN D M，SALIBA A，LOZANO D，et al. β-hydroxybutyrate enhances brain metabolism in normoglycemia and hyperglycemia，providing cerebroprotection in a mouse stroke model[J]. J Cereb Blood Flow Metab，2025，45（8）：
1493-1506.

β-羟基丁酸通过TRIB3/Akt/mTOR通路调控蛋白质合成和降解平衡改善卒中相关性肌少症

β-Hydroxybutyrate Improves Stroke-Related Sarcopenia by Regulating Protein Synthesis and Degradation Homeostasis via the TRIB3/Akt/mTOR Pathway

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