| Purpose:To investigate the damage of retina and PI3K/AKT(phosphatidylinositol 3-kinase/protein kinase B)after blast injury in mice and the expression of signal pathway and related apoptotic proteins.Method:Will be six to eight weeks of C57 mice abdominal cavity after anesthesia,in a high simulation model of blast injury device of the mesh,reveal other parts of the head and protect mice,mice below for air compressor,the thickness is about 0.8 mm aluminum thin in the middle tier of fixed,when the pressure reaches a critical level after the power supply to make the aluminum membrane rupture,gun barrel under the pressure of 0.1-0.14 MPa.Sixty male mice were randomly divided into control group and model group,and the model group was divided into 6 h group and 48h group according to the sampling time after successful modeling.The pathological changes of retina were observed by HE staining and the cell density of RPE and Inner Nuclear Layer(INL)were detected.The apoptosis of retina cells was detected by TUNEL assay,and the relative expression of p-PI3K,p-Akt,Bax,Bcl-2 and Cytochrome C in retina tissues were detected by Western Blot.SPSS25.0 software was used for statistical analysis of the data.T-test and chi-square test were used to complete the data between groups,and P<0.05 was considered statistically significant.Results:1.HE staining results:compared with the control group,the density of INL cells in the 6h and 48h groups was significantly decreased(both P<0.05);Compared with the 6h group,the INL cell density in 48h group was significantly decreased(all P<0.05);2.TUNEL staining results:Image J software was used to count the number of apoptotic cells and convert it into apoptotic rate.Compared with blank control group,the apoptosis rate of retinal cells in 6h and 48h groups was significantly increased(P<0.05).Compared with the 6h group,the apoptosis rate of retinal cells in the 48h group increased,and the difference was statistically significant.3.Western Blot results:The expressions of p-PI3K,p-Akt,Bax,Bcl-2 and Cytochrome C in 6-h and 48-h groups were significantly increased compared with blank control group(P<0.05).Compared with the 6-h group,the expression of p-PI3K,p-Akt and Cytochrome C in the 48-h group increased(P<0.05),and the expression of Bax/Bcl-2 in the 48-h group was not statistically significant(P>0.05).Conclusions:1.Detonation shock wave can cause retinal damage.In the model group,the number of cells in the retinal ganglion cell layer was significantly reduced and the cell arrangement was disordered,and the damage was aggravated with time.2.To observe the retinal apoptosis after shock wave exposure,TUNEL staining was performed on the retinal slices,and it was detected that the retinal positive cells in the model group were mainly distributed in the retinal ganglion cell layer and the outer nuclear layer.Apoptosis occurred in the ganglion cell layer and the outer nuclear layer 6 h after the shock wave and became more obvious 48 h after the shock wave.3.The relative expression levels of apoptotic proteins such as Bax and cytochrome C in the retinal tissues were significantly increased after exposure to shock wave,which proved the occurrence of retinal apoptosis,which was consistent with TUNEL staining results.The expression levels of phosphorylated PI3K and Akt proteins in retinal tissue were significantly higher than those in blank control group,which confirmed that the abnormal activation of PI3K/Akt signaling pathway in retina after knock shock,and the activation of PI3K/Akt signaling pathway was involved in the retinal injury and apoptosis process after knock injury. |
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