Histochemical analysis
The spinal cords dissected from the spinal canal were fixed in 4%
buffered paraformaldehyde for 24 h and then embedded in paraffin. In the
following step, the standard tissue sections (5 μm) were subjected to
hematoxylin and eosin (H&E) staining and luxol fast blue (LFB)
staining. Images were acquired using a digital color charge-coupled
device camera and NIS-Elements imaging software. The organ sections
(heart, liver, spleen, lung, and kidney) from vehicle- or piperine-
treated EAE mice was subjected to H&E analysis.