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Effects of Nogo-A Silencing on TNF-α and IL-6 Secretion and TH Downregulation in Lipopolysaccharide-Stimulated PC12 Cells.

Zhong J, Fan S, Yan Z, Xiao S, Wan L, Chen C, Zhong S, Liu L, Liu J - Biomed Res Int (2015)

Bottom Line: The viabilities of PC12 cells decreased with increase of LPS concentrations.LPS significantly increased the supernatant TNF-alpha and IL-6 concentrations and reduced TH protein expression in PC12 cells, while silencing Nogo-A could block these effects.These results suggested that LPS can activate PC12 cells to secrete inflammatory cytokines and lower the TH expression, which can be regulated by Nogo-A gene silencing.

View Article: PubMed Central - PubMed

Affiliation: Neurology Department, People's Hospital of Zengcheng City (Boji Hospital Affiliated to Sun Yat-sen University), Guangzhou 511300, China.

ABSTRACT
Parkinson's disease (PD) is a common degenerative disease that lacks efficient treatment. Myelin-associated neurite outgrowth inhibitor A (Nogo-A) is relevant with inhibition of nerve regeneration and may play vital role in pathogenesis of PD. The study aimed to establish the shRNA expression plasmids of Nogo-A gene and explore the regulatory effects of Nogo-A silencing on the expression of inflammation factor tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) as well as tyrosine hydroxylase (TH) in lipopolysaccharide- (LPS-) stimulated rat PC12 cells. The results showed that both mRNA and protein levels of Nogo-A in pGenesil-nogoA-shRNA group were downregulated. The viabilities of PC12 cells decreased with increase of LPS concentrations. LPS significantly increased the supernatant TNF-alpha and IL-6 concentrations and reduced TH protein expression in PC12 cells, while silencing Nogo-A could block these effects. These results suggested that LPS can activate PC12 cells to secrete inflammatory cytokines and lower the TH expression, which can be regulated by Nogo-A gene silencing. Nogo-A silencing might provide new ideas for PD treatment in the future.

No MeSH data available.


Related in: MedlinePlus

Cell viability tested by CCK8. (a) Gradual increase of LPS concentrations. (b) Silencing of Nogo-A on PC12 cells after treatment with LPS 1 nmol/L. ∗#p < 0.05.
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fig2: Cell viability tested by CCK8. (a) Gradual increase of LPS concentrations. (b) Silencing of Nogo-A on PC12 cells after treatment with LPS 1 nmol/L. ∗#p < 0.05.

Mentions: CCK8 tests showed that PC12 cell viabilities at hour 24 decreased gradually (p < 0.05) with the gradual increase of LPS concentrations (0.01 to 100 nmol/L, Figure 2(a)) and got a relatively ideal value at 1 nmol/L (76 ± 2.1%), which was used in subsequent experiments. Moreover, silencing of Nogo-A attenuated the viability loss of PC12 cells after LPS challenge (Figure 2(b)).


Effects of Nogo-A Silencing on TNF-α and IL-6 Secretion and TH Downregulation in Lipopolysaccharide-Stimulated PC12 Cells.

Zhong J, Fan S, Yan Z, Xiao S, Wan L, Chen C, Zhong S, Liu L, Liu J - Biomed Res Int (2015)

Cell viability tested by CCK8. (a) Gradual increase of LPS concentrations. (b) Silencing of Nogo-A on PC12 cells after treatment with LPS 1 nmol/L. ∗#p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4637059&req=5

fig2: Cell viability tested by CCK8. (a) Gradual increase of LPS concentrations. (b) Silencing of Nogo-A on PC12 cells after treatment with LPS 1 nmol/L. ∗#p < 0.05.
Mentions: CCK8 tests showed that PC12 cell viabilities at hour 24 decreased gradually (p < 0.05) with the gradual increase of LPS concentrations (0.01 to 100 nmol/L, Figure 2(a)) and got a relatively ideal value at 1 nmol/L (76 ± 2.1%), which was used in subsequent experiments. Moreover, silencing of Nogo-A attenuated the viability loss of PC12 cells after LPS challenge (Figure 2(b)).

Bottom Line: The viabilities of PC12 cells decreased with increase of LPS concentrations.LPS significantly increased the supernatant TNF-alpha and IL-6 concentrations and reduced TH protein expression in PC12 cells, while silencing Nogo-A could block these effects.These results suggested that LPS can activate PC12 cells to secrete inflammatory cytokines and lower the TH expression, which can be regulated by Nogo-A gene silencing.

View Article: PubMed Central - PubMed

Affiliation: Neurology Department, People's Hospital of Zengcheng City (Boji Hospital Affiliated to Sun Yat-sen University), Guangzhou 511300, China.

ABSTRACT
Parkinson's disease (PD) is a common degenerative disease that lacks efficient treatment. Myelin-associated neurite outgrowth inhibitor A (Nogo-A) is relevant with inhibition of nerve regeneration and may play vital role in pathogenesis of PD. The study aimed to establish the shRNA expression plasmids of Nogo-A gene and explore the regulatory effects of Nogo-A silencing on the expression of inflammation factor tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) as well as tyrosine hydroxylase (TH) in lipopolysaccharide- (LPS-) stimulated rat PC12 cells. The results showed that both mRNA and protein levels of Nogo-A in pGenesil-nogoA-shRNA group were downregulated. The viabilities of PC12 cells decreased with increase of LPS concentrations. LPS significantly increased the supernatant TNF-alpha and IL-6 concentrations and reduced TH protein expression in PC12 cells, while silencing Nogo-A could block these effects. These results suggested that LPS can activate PC12 cells to secrete inflammatory cytokines and lower the TH expression, which can be regulated by Nogo-A gene silencing. Nogo-A silencing might provide new ideas for PD treatment in the future.

No MeSH data available.


Related in: MedlinePlus