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Picroside II protects neurons from inflammation by inhibiting p38-p53 signaling pathway in vivo and in vitro

Wang Tingting  Li Shan  Zhao Li  Zhai Li  Zhang Yanxue  Guo Yunliang  
【摘要】:Objective To explore the neuroprotective effect of picroside II on p38 signalling pathway after cerebral ischemia/reperfusion injury in vivo and in vitro.Methods In vivo, the focal cerebral ischemic models were established by inserting a monofilament threads into middle cerebral artery. All rats were randomly divided into sham group, model group and picroside(Picr) group. The neurobehavioral function was evaluated by modified neurological severity score points test. The structure of neuron was observed using hematoxylin-eosinstaining staining and transmission electron microscopy. The apoptotic cells were counted using terminal deoxynucleotidyl transferase dUTP nick end labeling assay. The expression of phosphorylated p38(p-p38) in cortex was determined using the immunohistochemistry. And the expressions of p-p38, phosphorylated p53(p-p53),interleukin-6(IL-6) and tumor necrotic factor α(TNFα) were determined by Western Blot. In vitro, SH-SY5 Y cells were used to establish the oxygen glucose deprivation/reoxygen(OGD/R) model, which was divided into the control group,model group, Picr group, anisomycin(Anis) group, Anis+Picr group, SB203580(SB)group, and SB+Picr group. Cell morphology was observed under light microscopy.Cell viability was detected by CCK8. Apoptoic cells were detected by flow cytometry,and expressions of p-p38, p-p53, IL-6 and TNFα were detected by immunofluorescence and Western Blot. Results In model group, the damage of neuron was worsened, while the neurobehavioral function score, apoptotic cell index and the expressions of p-p38, p-p53, IL-6 and TNFα increased significantly than those in control group. In Picr group, the damage of neuron was lighter, the neurological behavioral function was improved, the number of apoptotic cells and the expressions of p-p38, p-p53 and IL-6 decreased significantly than those in model group. In vitro experiments, 5 Y cells showed shrinkage, decreased survival rate, increased apoptotic cells, and enhanced protein expressions following OGD/R. In Picr, SB and SB+Picr groups, the damage of 5 Y cells was lighter, the number of apoptotic cells and the expressions of p-p38, p-p53 and IL-6 decreased significantly than those in model group. In Anis and Anis+Picr groups, the damage was worsen, and the expressions of p-p38, p-p53, IL-6 and TNFα increased, which were similar to the model group.Conclusion Picroside II could protect the neuron from the apoptosis and inflammation reaction after MCAO/R and OGD/R by inhibiting p38-p53 signaling pathway.

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