Gene Expression Profiling Analysis to Investigate the Role of CIMT after Stroke in Neurological Behaviors

Objectives: Constraint induced movement therapy (CIMT) promoting neurogenesis, synaptic plasticity and axon regeneration after stroke in animal models, but the candidate genes and pathways associated with these changes are unknown. The present study was conducted to explore the effects of CIMT on motor function and cognitive function in stroke-induced rats. Methods The study included three groups: sham-operated rats (n = 6, SHAM), rats subjected to cerebral ischemia (n = 8, ISC), rats with cerebral ischemia plus CIMT(n = 8, CIMT). A cerebral infarction rat model was established by transient middle cerebral artery occlusion (tMCAO). CIMT began on the 8th day after the cerebral ischemia surgery by fitting a plaster cast around the unimpaired upper limbs of rats for 3 weeks. The neurological scoring test was performed to evaluate the neurological function of rats. Stable Platform Test and Coat Hanger Test were performed to evaluate the motor function. The Morris water maze(MWM)test was conducted to assess the cognitive function of rats. RNA sequencing and bioinformatics analysis were employed to study the underlying mechanisms. Results We observed that CIMT significantly ameliorate the memory dysfunction as well as neurological deficits in cerebral ischemic rats caused by tMCAO. We identified a total of 105 DEGs, including 71 upregulated genes and 34 downregulated genes. GO analysis revealed that these DEGs were associated with the major categories: at the MF level, the DEGs were mainly enriched in N-acetylgalactosamine 4-O-sulfotransferase activity, lyase activity, glutathione transferase activity and oxidoreductase activity, acting on paired donors. KEGG analysis showed that the DEGs were mainly enriched in 4 categories: environmental information processing, human disease, metabolism, organismal system. Conclusions We identified the following 16 genes with the most significant differential expression: En1, Cpne7, Siglec5, Chchd4, Fcrl2, Trhr, Myom3, Piezo2, Ankrd34b, Slc6a3, Rftn1, Chst9, Gstm6, Cd34 and Tgm2. These findings prompt exploration of the potential mechanisms and key genes involved in the effect of CIMT.