Fig. 6
Intrathecal BMSCs attenuate the upregulation of P2X4R provoked by CCD. a Western blot analysis of P2X4R proteins in the membrane fraction from the spinal cord ipsilateral to surgery at different time points in sham and CCD rats. b BMSC injection downregulated the level of P2X4R protein at days 6 post-CCD. c Primary microglia were incubated with BMSC lysate for 24 h, and the upregulation of P2X4R expression induced by FN (10 μg/mL) was suppressed. GAPDH served as a loading control. Each time point had a sample size of 6 and data are presented as the mean percentage of each control (SEM). d P2X4R immunofluorescence was barely detectable in sham animals. e At 6 days post-CCD, P2X4R immunoreactivity was clearly induced in the dorsal horn ipsilateral to the lesion side (left top and left bottom); this signal was weaker in the lumbar spinal cords from BMSC graft animals (right top and right bottom). Of note, microglia remain the dominant population in the dorsal horn for both groups (demarcated by the red line), suggesting that BMSC treatment did not affect the number of microglia residing in this area. f At 14 days post-CCD, there was no difference in the P2X4R immunoreactivity between the CCD rats (left top and bottom) and BMSC-treated (right top and bottom). Bottom photography in e and f are magnified images of their corresponding parent image above (area magnified demarcated by a blue square). **p < 0.01