Therapeutic mechanism of transcranial iTBS on nerve regeneration and functional recovery in rats with complete spinal cord transection

Abstract

Background: After spinal cord transection injury, the inflammatory microenvironment formed in the injury site and the cascade of secondary injury results in limited regeneration of injured axons and the apoptosis of neurons in the sensorimotor cortex (SMC). It is crucial to reverse these adverse processes for the recovery of voluntary movement. In this study, transcranial intermittent theta-burst stimulation (iTBS) was used for the treatment of complete spinal cord transection in rats. The mechanism of transcranial iTBS as a new non-invasive neural regulation paradigm in promoting axonal regeneration and motor function repair was explored. Methods: Rats from the iTBS group were treated with transcranial iTBS 72h after spinal cord injury (SCI). Each rat was received behavioral testing. Inflammation, neuronal apoptosis, neuroprotective effect, regeneration and synaptic plasticity were measured by immunofluorescence staining, western blotting and mRNA sequencing 2 or 4w after SCI. Each rat was received anterograde tracings in the SMC or the long descending propriospinal neurons and tested for motor evoked potentials. Regeneration of corticospinal tract (CST) and 5-hydroxytryptamine (5-HT) nerve fibers were detected eight weeks after SCI. Results: Compared with the control group and the sham iTBS group, rats of the iTBS group showed reduced inflammatory responses and neuronal apoptosis in the SMC two weeks after treatment. After four weeks, the neuroimmune microenvironment at the injury site was improved, and neuroprotective effects were seen to promote axonal regeneration and synaptic plasticity. Significantly, eight weeks after treatment, transcranial iTBS also increased the regeneration of CST, 5-HT nerve fibers, and the long descending propriospinal tract (LDPT). Moreover, motor evoked potentials and hindlimb motor function were significantly improved at eight weeks. Conclusions: Collectively, our results verified that iTBS has the potential to provide neuroprotective effects at early injury stages and pro-regeneration effects related to the 1) CST–5-HT; 2) CST–LDPT; and 3) CST–5-HT–LDPT descending motor pathways and revealed the relationships among neural pathway activation, neuroimmune regulation, neuroprotection, and axonal regeneration, as well as the interaction network of key genes. The proposed non-invasive transcranial iTBS treatment is expected to provide a serviceable practical and theoretical support for spinal cord injury.