Objective To explore the effect and mechanism of down-regulation of Toll like receptor 4(TLR4) mRNA on dendritic cell in hand-foot-mouth disease (HFMD) mice model,so as to provide reference for the prevention and treatment of HFMD. Methods Forty-five 12-day-old BALB/c suckling mice of SPF grade were selected to isolate and culture the dendritic cells.TLR4 siRNA sequence was designed and synthesized for cell infection.The suckling mice model of HFMD was established and divided into model group,down-regulated group and up-regulated group,with 15 animals in each group.Mice in the down-regulated group were intraperitoneally injected with dendritic cells that down-regulated TLR4 gene,and those in the up-regulated group were intraperitoneally injected with dendritic cells that up-regulated TLR4 gene,and the mice in the model group were intraperitoneally injected with the same amount of sterilized water.The proliferation and apoptosis of dendritic cells,the levels of serum interleukin-6 (IL-6),interleukin-12 (IL-12),tumor necrosis factor-α (TNF-α) and the expression of mitogen activated protein kinase (MAPK) pathway protein were detected and were compared among three groups. Results The proliferation rates of dendritic cells in the down-regulated group were lower than those in the up-regulated group at different time points (t=20.160,19.050,55.380,P<0.001).The apoptosis rates of dendritic cells in the down-regulated group at different time points were higher than those in the up-regulated group (t=34.740,84.570,48.020,P<0.001).The serum levels of IL-6,IL-12 and TNF-α in the down-regulated group were lower than those in the up-regulated group(t=39.330,12.660,33.690,P＜0.001).The expression levels of P-P38MAPK,p-JNK and p-ERK1/2 protein in the down-regulated group were lower than those in the up-regulated group (t=33.690,17.470,27.800,P＜0.001). Conclusion Down regulation of TLR4 mRNA can down regulating the expression of IL-6,IL-12 and TNF-α through inhibiting MAPK pathway activation and regulating pathway protein,thereby promoting the apoptosis of dendritic cells in HFMD mice.
Key words
Toll like receptor /
hand foot mouth disease /
dendritic cell
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