Joint Therapy by NT-3-HUMSCs and SOCS3 Gene Dilencing in Nerve Regeneration Repair after Spinal Cord Injury in SD Rats / 昆明医科大学学报

ABSTRACT

Objective To investigate the effect of joint therapy by NT-3-HUMSCs and SOCS3 gene silencing in promoting the injury nerve regeneration repair after spinal cord injury in SD rats. Methods (1) Adherence method was used to culture human umbilical cord-derived mesenchymal cells (HUMSC) in vitro for separation, purification and identification. (2) We constructed NT-3 gene eukaryotic expression vector, and used gene transfection technology into its HUMSC, and tested the survival of NT-3-HUMSC cells and NT-3 expression in cells. (3) We screened specific targets of SOCS3, made sequence homology analysis, and set a negative control, designed and synthesized siRNA and detected the function. (4) SD rats model of spinal cordinjury were established and divided into 1. sham group 10; 2.T12 whole spinal cord injury model 40, were randomly divided into four groups, respectively; saline treatment group 10; siRNA + NT-3-HUMSCs treatment group 10; NT-3-HUMSCs treatment group 10; siRNA treated group 10. After each group above modeling success, they received respectively the neural electrophysiological monitoring for 12 weeks survival. (5) We perfused SD rats for fixation and collect samples, and observed the local glial scar degradation situation and axon regeneration, meanwhile, used biotin glucan fluorescent (BDA) anterograde tracing. The injury transplant area-host junction spinal cord tissues were collected to observe the corticospinal tract regeneration under microscope. Results (1) In siRNA + NT-3-HUMSCs treatment group, the transection syringomyelia was significantly reduced as compared with normal saline group (P ＜ 0.05). (2) BDA anterograde tracing results showed that in the siRNA + NT-3-HUMSCs treatment group, neural axon grew significantly compared with the normal saline group. (3) Neural electrophysiological testing 12 weeks after injury in the treatment group, the incubation period P40 was shorter as compared with control group; in siRNA + NT-3-HUMSCs treatment group, the incubation period was shorter obviously than normal saline, but the amplitude increased obviously (P ＜ 0.05). Conclusion NT-3-HUMSCs joint with SOCS3 gene silencing can promote the injury nerve regeneration repair in the treatment of SD rat spinal cord injury.