Objective    To investigate the relationship between p38MAPK and astrocytes activation by detecting the changes of p38MAPK

TNF-α

NO and NOS in the substance P-induced activation of spinal cord astrocytes

in vitro

.      Methods    Astrocytes

cultured from spinal cord of SPF rat and identified by immunofluorescence method with GFAP antibody

were grouped into the control group

the SP stimulus group (SP group) and the SB20

3580 interrupt p38MAPK group (interrupt group) in which SP (10

-7

mol/L) and SB203580 (10 μmol/L) were added to the supernatant for 12 h. The WB method

RT-PCR and ELISA were used to determine the changes of p38MAPK

phosphorylated p38MAPK

GFAP mRNA

TNF-α

NO and NOS in the astrocytes or supernatant at 1

3

12

24 and 36 h respectively.     Results    The GFAP positive rate of the cultured cells was higher than 95%. p38MAPK level in the SP group did not have any change

whereas p-p38 level increased significantly after 1 h and GFAP mRNA

NO

NOS and TNF-α level increased after 3 h. When p38MAPK pathway was inhibited by SB203580 in the SP+SB group

GFAP mRNA

NO

NOS and TNF-α were significantly reduced compared with those in the SP group. The level of p38MAPK

p-p38

GFAP mRNA

NO

NOS and TNF-α in the SB group did not have changes compared with those in the normal group.     Conclusion    These results indicate that p38MAPK signal pathway contributes to substance P induced activation of spinal cord astrocytes according to inflammatory factors attenuation after p38MAPK signal pathway interruption by SB203580.