Ginsenoside Rg1 Regulates The Expression Of Surface Proteins On Astrocyte And Its Effects In Spinal Cord Injury

Objective To observe the effect of Ginsenoside Rgl intervention on surface proteins(nestin,GLT1,NG2,AQP4,GFAP,Vimentin)expression on astrocytes(AS)and to explore its possible molecular mechanism and the protection effect of Ginsenoside Rgl on the spinal cord(SCI),proposing a novel strategy for treating SCIMethods Astrocytes were primary cultured and purified in vitro from the brain of 24 h SD rats;Immunofluorescence was used to detect the purity of astrocytes,and more than 95%of the cells were used for subsequent experiments.CCK-8 assay was used to detect the effect of ginsenoside Rgl on the activity of AS cells after scratch under the concentration gradient of 10,20,40,80 ?g/ml.Optimal concentration of ginsenoside Rgl was determined and performed in the succedent experiments.The scratch model of astrocytes in vitro was established by scratch method,and purified cells were divided into 2 groups,the experimental group(Rg 1 group)were treated with Ginsenoside Rgl(the optimal concentration),and the control group was only treated with medium.The morphological changes and scratch healing speed after cell injury in the two groups were observed and compared.The effect of Rg1 on the expression of surface proteins(Nestin,GLT1,NG2,AQP4,GFAP,Vimentin)on astrocyte scratch cultured in vitro was investigated by Western-blot.The protein levels of phosphatidylinositol 3-kinase(PI3K)?phospho-PI3K(p-PI3K)?protein kinase B(Akt)?phospho-Akt(p-Akt)were examined by western blot in Rgl treated injured AS after intervention of PI3K/Akt inhibitor LY294002Results The astrocytes cultured in vitro had normal morphology and good activity,and the purity reached more than 95%identificated by immunofluorescence,which can be used in the following experiments;Cell Counting Kit-8(CCK-8)assay showed that the activity of scratch cells was the highest when Rgl concentration was 40 ?g/ml(P<0.05).Scratch method indyced the injury model of astrocytes in vitro successfully,and caused significant morphological changes of astrocytes observed by inverted microscope.Quantitative statistical analysis showed that the healing rate of Rg1 group was faster than that of control group after 24 h(P<0.05).Western-blot analysis demonstrated that expressions of Nestin?GLT1?NG2 were significantly up-regulated in ginsenoside Rgl group compared to those in control group(P<0.05),while the expression of AQP4,GFAP,Vimentin in the Rg1 group was relatively down-regulated(P<0.05).After LY294002 intervention,the relative protein expression of P-PI3K/PI3K and p-Akt/Akt,and the difference was statistically significant(P<0.05)compared with the control group;While the expression of P-PI3K/PI3K and p-Akt/Akt after PI3K/Akt entry inhibitor LY294002 was significantly inhibited,compared with Rgl group(P<0.05).Conclusions The vitro experiments showed that ginsenoside Rg1 may positively regulate the surface antigens on astrocytes(Nestin,GLT1,NG2,AQP4,GFAP,Vimentin)to promote the repair of scratch injury,and hence benefits to spinal cord injury repair,and PI3K/Akt signaling pathways may be involved in the process.