ARNT调控低氧通路促进小鼠骨骼肌再生的实验研究

doi:10.13418/j.issn.1001-165x.2021.06.011

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中国临床解剖学杂志 ›› 2021, Vol. 39 ›› Issue (6) : 673-679. DOI: 10.13418/j.issn.1001-165x.2021.06.011

实验研究

ARNT调控低氧通路促进小鼠骨骼肌再生的实验研究

李玢，　张誉腾，　鲁峰

作者信息 +

Study on aryl hydrocarbon receptor nuclear translocator regulating hypoxic pathway to promote skeletal muscle regeneration in mice

Li Bin, Zhang Yuteng, Lu Feng

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文章历史 +

摘要

目的　探索芳香烃受体核转运子（aryl hydrocarbon receptor nuclear translocator，ARNT）对骨骼肌再生的调控作用，为提高老龄人口骨骼肌再生能力提供方向。方法　构建小鼠骨骼肌冰冻损伤模型，观察幼龄鼠与老龄鼠、骨骼肌特异性ARNT基因敲除小鼠与野生鼠、加入低氧通路激活剂（ML228）与DMSO老龄鼠间骨骼肌再生能力差别；分析ARNT、低氧通路、肌再生因子表达含量及小鼠下肢血流差异。结果　衰老导致骨骼肌再生能力减弱；敲除ARNT基因后，小鼠骨骼肌的再生能力显著下降，低氧通路因子及相关基因表达下调；ML228可使受损的骨骼肌再生能力得到恢复。结论　衰老引起的ARNT含量下降抑制低氧通路是导致老龄骨骼肌再生能力降低的主要原因。低氧通路激活剂可改善受损骨骼肌的再生能力，有望成为药物重塑衰老骨骼肌再生能力的新靶点。

Abstract

Objective　To explore the regulatory role of aryl hydrocarbon receptor nuclear transporter (ARNT) on skeletal muscle regeneration and to provide theories for improving muscle regeneration in aging population.　Methods　Mice skeletal muscle injury model was constructed. Skeletal muscle regeneration ability was explored between young and old mice, transgenic and wild-type mice, old hypoxia pathway activator (ML228) and DMSO mice following cryoinjury. Whole muscle was obtained and analyzed to evaluate the expressions of ARNT, hypoxia pathway and skeletal muscle regeneration related protein/ genes, as well as the blood flow in the lower limbs of mice.　Results　The regeneration ability of skeletal muscle decreased in aging mice. ARNT gene knockout in young mice hindered skeletal muscle regeneration, as well as down-regulated hypoxia pathway factor and the expressions of related genes. ML228 could restore the regenerative ability of damaged skeletal muscle.　Conclusions　The inhibition of hypoxia pathway by the decreasing of ARNT content caused by aging is the main reason for the decreasing of skeletal muscle regeneration ability. The pharmacological activator of hypoxia signal improves the regeneration ability of skeletal muscle in the elderly is promising.

导出引用

李玢, 张誉腾, 鲁峰. ARNT调控低氧通路促进小鼠骨骼肌再生的实验研究[J]. 中国临床解剖学杂志. 2021, 39(6): 673-679 https://doi.org/10.13418/j.issn.1001-165x.2021.06.011

Li Bin, Zhang Yuteng, Lu Feng. Study on aryl hydrocarbon receptor nuclear translocator regulating hypoxic pathway to promote skeletal muscle regeneration in mice[J]. Chinese Journal of Clinical Anatomy. 2021, 39(6): 673-679 https://doi.org/10.13418/j.issn.1001-165x.2021.06.011

中图分类号： R337

参考文献

[1] Aärimaa V, Kääriäinen M, Vaittinen S, et al. Restoration of myofiber continuity after transection injury in the rat soleus[J]. Neuromuscul Disord, 2004, 14(7): 421-428. DOI: 10.1016/j.nmd.2004.03.009.
[2] Majmundar AJ, Lee DSM, Skuli N, et al. HIF modulation of Wnt signaling regulates skeletal myogenesis in vivo[J]. Development, 2015, 142(14): 2405-2412. DOI: 10.1242/dev.123026.
[3] Yang X, Yang SQ, Wang C, et al. The hypoxia-inducible factors HIF1α and HIF2α are dispensable for embryonic muscle development bur are essential for postnatal muscle regeneration[J]. J Biol Chem, 2017, 292(14): 5981-5991. DOI: 10.1074/jbc.M116.756312.
[4] Lee Z, Dennis J, Alsberg E, et al. Imaging stem cell differentiation for cell-based tissue repair[J]. Methods Enzymol, 2012, 506: 247-263. DOI: 10.1016/B978-0-12-391856-7.00037-8.
[5] Urban JD, Budinsky RA, Rowlands JC. Single nucleotide polymorphisms in the human aryl hydrocarbon receptor nuclear translocator (ARNT) gene[J]. Drug Metab Pharmacokinet, 2011, 26(6): 637-645. DOI: 10.2133/dmpk.DMPK-11-SC-031.
[6] Chan YY, Kalpana S, Chang WC, et al. Expression of aryl hydrocarbon receptor nuclear translocator enhances cisplatin resistance by upregulating MDR1 expression in cancer cells[J]. Mol Pharmacol, 2013, 84(4): 591-602. DOI: 10.1124/mol.113.087197.
[7] Obacz J, Pastorekova S, Vojtesek B, et al. Cross-talk between HIF and p53 as mediators of molecular responses to physiological and genotoxic stresses[J]. Mol Cancer, 2013, 12(1): 93. DOI: 10.1186/1476-4598-12-93.
[8] Tomita S, Sinal CJ, Yim SH, et al. Conditional disruption of the aryl hydrocarbon receptor nuclear translocator (Arnt) gene leads to loss of target gene induction by the aryl hydrocarbon receptor and hypoxia-inducible factor 1alpha[J]. Mol Endocrinol, 2000, 14(10): 1674-1681. DOI: 10.1210/mend.14.10.0533.
[9] Theriault JR, Felts AS, Bates BS, et al. Discovery of a new molecular probe ML228: an activator of the hypoxia inducible factor (HIF) pathway[J]. Bioorg Med Chem Lett, 2012, 22(1): 76-81. DOI: 10.1016/j.bmcl.2011.11.077.
[10] Chen H, Li J, Liang S, et al. Effect of hypoxia‑inducible factor‑1/vascular endothelial growth factor signaling pathway on spinal cord injury in rats[J]. Exp Ther Med, 2017, 13(3): 861-866. DOI: 10.3892/etm.2017.4049.