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.