颅神经损伤面颈部肌萎缩纤维化中环状RNA差异表达分析

Analysis of Differential Expression of Circular RNA in Muscular Atrophy and Fibrosis of Facial and Neck Cranial Nerve Injury

高轶昳;刘菲;郑宏良;周义德;陈世彩;黄卫;于阅尽;郝亚楠;

1:海军军医大学附属长海医院耳鼻咽喉科

2:上海市中医药大学附属第七人民医院耳鼻咽喉科

摘要
目的检测颅神经损伤后面颈部骨骼肌萎缩纤维化过程中环状RNA(circular RNA,circRNA)的差异表达,探讨circRNA在面颈部失神经骨骼肌萎缩纤维化中的潜在功能。方法使用12周龄C57BL/6雄性小鼠共50只,分为对照组(仅行右侧副神经探查术)、失神经支配1周组、失神经支配2周组、失神经支配3周组及失神经支配4周组,每组10只;失神经支配各组建立颅神经损伤失神经支配肌萎缩纤维化模型(均切除5 mm右侧副神经)。各组分别取术侧胸锁乳突肌进行Masson染色观察骨骼肌形态,高通量转录组测序技术(RNA-seq)检测各组小鼠失神经支配胸锁乳突肌的circRNA差异表达,并行生物信息学分析。结果 Masson染色验证骨骼肌组织萎缩纤维化显著,造模成功;鉴定失神经骨骼肌具有差异性表达的circRNA共1 386个,其中上调686个,下调700个,与对照组比较,失神经支配1、2、3、4周组分别上调83、138、314、151个,下调103、149、204、244个;GO功能分析显示,这些差异表达circRNA的亲本基因在失神经损伤早期与肌细胞分化和发育等相关,失神经损伤晚期则与肌细胞凋亡、结缔组织增生等相关。KEGG分析显示TGF-β、Apelin信号通路、FoxO信号通路等与纤维化调控密切相关的信号通路在各实验组均有不同程度的富集。结论面颈部颅神经损伤肌萎缩纤维化过程中circRNA具有差异性表达,可能参与喉返神经等颅神经的失神经肌萎缩纤维化进程。
关键词
失神经支配;纤维化;肌萎缩;环状RNA
基金项目(Foundation):
国家自然科学基金项目(81700897、81970868)
作者
高轶昳;刘菲;郑宏良;周义德;陈世彩;黄卫;于阅尽;郝亚楠;
参考文献

1 Liu F,Tang W,Chen D,et al.Expression of TGF-β1 and CTGF is associated with fibrosis of denervated sternocleidomastoid muscles in mice[J].The Tohoku Journal of Experimental Medicine,2016,238(1):49-56.

2 Wilusz JE,Sharp PA.Molecular biology.A circuitous route to noncoding RNA[J].Science,2013,340(6131):440-441.

3 Qin C,Liu CB,Yang DG,et al.Circular RNA expression alteration and bioinformatics analysis in rats after traumatic spinal cord injury[J].Front Mol Neurosci,2019,11:497.doi.org/10.3389/fnmol.2018.00497.

4 Mao S,Zhang S,Zhou S,et al.A Schwann cell-enriched circular RNA circ-Ankib1 regulates Schwann cell proliferation following peripheral nerve injury[J].FASEBJ,2019,33(11):12409-12424.

5 Wang Y,Li M,Wang Y,et al.A Zfp609 circular RNA regulates myoblast differentiation by sponging miR-194-5p[J].Int J Biol Macromol,2019,121:1308-1313.doi:10.1016/j.ijbiomac.2018.09.039

6 Li H,Wei X,Yang J,et al.circFGFR4 promotes differentiation of myoblasts via binding miR-107 to relieve its inhibition of Wnt3a[J].Mol Ther Nucleic Acids,2018,11:272-283.doi:10.1016/j.omtn.2018.02.012.

7 Suzuki H,Aoki Y,Kameyama T,et al.Endogenous multiple exon skipping and back-splicing at the DMD mutation hotspot[J].Int J Mol Sci,2016,17(10):1722.doi:10.3390/ijms17101722.

8 Li J,Li P,Zhang G,et al.CircRNA TADA2A relieves idiopathic pulmonary fibrosis by inhibiting proliferation and activation of fibroblasts[J].Cell Death Dis,2020,11(7):553.

9 Ji D,Chen GF,Wang JC,et al.Hsa_circ_0070963 inhibits liver fibrosis via regulation of miR-223-3p and LEMD3[J].Aging (Albany NY),2020,12(2):1643-1655.

10 Batt J,Bain J,Goncalves J,et al.Differential gene expression profiling of short and long term denervated muscle[J].FASEB J,2006,20(1):115-117.

11 Quan G,Li J.Circular RNAs:biogenesis,expression and their potential roles in reproduction[J].J Ovarian Res,2018,11(1):1-12.

12 Sohn EJ,Park HT.Differential expression of circular RNAs in the proximal and distal segments of the sciatic nerve after injury[J].Neuroreport,2020,31(1):76-84.

13 Wang Y,Li M,Wang Y,Liu J,et al.A Zfp609 circular RNA regulates myoblast differentiation by sponging miR-194-5p[J].Int J Biol Macromol,2019,121:1308-1313.doi.org/10.1016/j.ijbiomac.2018.09.039

14 Wei X,Li H,Yang J,et al.Circular RNA profiling reveals an abundant circLMO7 that regulates myoblasts differentiation and survival by sponging miR-378a-3p[J].Cell Death Dis,2017,8(10):e3153.dx.doi.org/10.1038/cddis.2017.541.

15 Chen Y,Yuan B,Wu Z,et al.Microarray profiling of circular RNAs and the potential regulatory role of hsa_circ_0071410 in the activated human hepatic stellate cell induced by irradiation[J].Gene,2017,629:35-42.doi.org/10.1016/j.gene.2017.07.078.

16 Zhou B,Yu JW.A novel identified circular RNA,circRNA_010567,promotes myocardial fibrosis via suppressing miR-141 by targeting TGF-β1[J].Biochem Biophys Res Commun,2017,487(4):769-775.

17 Yang X,Wang J,Zhou Z,et al.Silica-induced initiation of circular ZC3H4 RNA/ZC3H4 pathway promotes the pulmonary macrophage activation[J].FASEB J,2018,32(6):3264-3277.

18 Ferrié C,Kasper S,Wanivenhaus F,et al.Focal adhesion kinase coordinates costamere-related JNK signaling with muscle fiber transformation after Achilles tenotomy and tendon reconstruction[J].Exp Mol Pathol,2019,108:42-56.doi.org/10.1016/j.yexmp.2019.03.006.

19 Hu HH,Chen DQ,Wang YN,et al.New insights into TGF-β/Smad signaling in tissue fibrosis[J].Chem Biol Interact,2018,292:76-83.doi.org/10.1016/j.tice.2019.01.003.

20 Doan A.Apelin receptor (Aplnr) signaling promotes fibroblast migration[J].Tissue Cell,2019,56:98-106.doi.org/10.1016/j.tice.2019.01.003.

21 Schiaffino S,Dyar KA,Ciciliot S,et al.Mechanisms regulating skeletal muscle growth and atrophy[J].FEBS J,2013,280(17):4294-4314.