Lin Yao-Fa, Xie Zheng, Zhou Jun, Yin Gang, Lin Hao-Dong
Department of Orthopedic Surgery, Changzheng Hospital, the Second Military Medical University, Shanghai, China.
Department of Orthopedic Surgery, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Neural Regen Res. 2019 Dec;14(12):2183-2191. doi: 10.4103/1673-5374.262602.
Wallerian degeneration and nerve regeneration after injury are complex processes involving many genes, proteins and cytokines. After different peripheral nerve injuries the regeneration rate can differ. Whether this is caused by differential expression of genes and proteins during Wallerian degeneration remains unclear. The right tibial nerve and the common peroneal nerve of the same rat were exposed and completely cut through and then sutured in the same horizontal plane. On days 1, 7, 14, and 21 after surgery, 1-2 cm of nerve tissue distal to the suture site was dissected out from the tibial and common peroneal nerves. The differences in gene and protein expression during Wallerian degeneration of the injured nerves were then studied by RNA sequencing and proteomic techniques. In the tibial and common peroneal nerves, there were 1718, 1374, 1187, and 2195 differentially expressed genes, and 477, 447, 619, and 495 differentially expressed proteins on days 1, 7, 14, and 21 after surgery, respectively. Forty-seven pathways were activated during Wallerian degeneration. Three genes showing significant differential expression by RNA sequencing (Hoxd4, Lpcat4 and Tbx1) were assayed by real-time quantitative polymerase chain reaction. RNA sequencing and real-time quantitative polymerase chain reaction results were consistent. Our findings showed that expression of genes and proteins in injured tibial and the common peroneal nerves were significantly different during Wallerian degeneration at different time points. This suggests that the biological processes during Wallerian degeneration are different in different peripheral nerves after injury. The procedure was approved by the Animal Experimental Ethics Committee of the Second Military Medical University, China (approval No. CZ20160218) on February 18, 2016.
损伤后的华勒氏变性和神经再生是涉及许多基因、蛋白质和细胞因子的复杂过程。不同的周围神经损伤后,再生速度可能不同。目前尚不清楚这是否是由于华勒氏变性过程中基因和蛋白质的差异表达所致。暴露同一大鼠的右胫神经和腓总神经,并完全切断,然后在同一水平面上进行缝合。术后第1、7、14和21天,从胫神经和腓总神经中切取缝合部位远端1 - 2 cm的神经组织。然后通过RNA测序和蛋白质组学技术研究损伤神经在华勒氏变性过程中的基因和蛋白质表达差异。在胫神经和腓总神经中,术后第1、7、14和21天分别有1718、1374、1187和2195个差异表达基因,以及477、447、619和495个差异表达蛋白质。华勒氏变性过程中有47条信号通路被激活。通过实时定量聚合酶链反应检测了3个经RNA测序显示有显著差异表达的基因(Hoxd4、Lpcat4和Tbx1)。RNA测序和实时定量聚合酶链反应结果一致。我们的研究结果表明,在不同时间点的华勒氏变性过程中,损伤的胫神经和腓总神经中的基因和蛋白质表达存在显著差异。这表明损伤后不同周围神经的华勒氏变性生物学过程不同。该实验程序于2016年2月18日获得中国第二军医大学动物实验伦理委员会批准(批准号:CZ20160218)。