Flow Cytometry
The expression levels of the M1 (anti-CD68) and M2 (anti-CD163) in the microglial cell of the L3-L5 segments of the spinal cord tissues were analyzed by the flow cytometry (Cytomics FC 500, Beckman, USA). The pancreatin (Beyotime, Shanghai, China) digested the spinal cord tissues at 37℃ for 5 min. The cell suspension was centrifuged at 1,000 rpm for 5 min and then transferred to a 1.5 mL EP tube. The total number of cells was 1×106/tube. The anti-CD68 (1:500, Abcam) and anti-CD163 (1:60, Abcam) were incubated at room temperature for 30 min in the dark. Finally, the expression levels of the anti-CD68 and anti-CD163 were detected by flow cytometer (Cytomics FC 500, Beckman).