Abstract: 【Objective】 To construct the small hairpin RNA(shRNA) expression plasmid targeting myelin associated inhibitor A(Nogo -A) and study the effect of Nogo -A gene shRNA on lipopolysaccharide (LPS) induced the release of inflammatory cytokines tumor necrosis factor alpha (TNF - alpha) and interleukin -6 (IL -6) in PC12 cells

and the expression of tyrosine hydroxylase (TH). 【Methods】 Nogo -A gene shRNA plasmid was constructed. Fluorescence Quantitative Real -Time PCR and Western blot were used to detect the expression of Nogo -A. PC12 cells were treated with different concentration of LPS for 24 hours and detected with cell viability assay CCK8. An appropriate concentration of LPS was chosen for the following experiment. PC12 cells were divided into control group

pGenesil -1.1+LPS group and pGenesil -nogoA -shRNA+LPS group . The content of the supernatant TNF - alpha and IL -6 of each group was determined by ELISA assay. Western blot was used for detection of TH. 【Results】 shRNA expression plasmid targeting Nogo -A was successfully constructed. Both mRNA and protein level of Nogo -A in pGenesil -nogoA -shRNA group were decreased

the difference is statistically significant (P < 0.05). The viability of PC12 cells decreased when LPS concentration increased. Compared with the control group

the content of TNF- alpha and IL-6 in the supernatant of LPS group was increased

while the TH level decreased. Compared with the LPS group

the content of TNF- alpha and IL-6 in the supernatant of pGenesil-nogoA-shRNA+LPS group decreased significantly

while the TH level increased

the difference of both comparisons were statistically significant (P < 0.05). 【Conclusion】 LPS induced PC12 cells to release inflammatory cytokines and lower the TH expression

while these effects can be regulated by silencing of the Nogo-A gene.