The Effect Of Quercetin,Oleanolic Acid And Icariin And Their Compatibility On The Apoptosis And P38MAPK And JNK Signaling Pathways In Hippocampal Neurons Of SD Rats Cultured With High Glucose Medium

[Objective]To explore the effect of quercetin,oleanolic acid and icariin and their compatibility on the apoptosis of hippocampal neurons of Sprague Dawley rats cultured with high glucose medium and the p38MAPK and JNK signaling pathway.[Methods]Hippocampus were obtained from newborn 24h Sprague Dawley rats,then separated and purified by trypsin digestion,gradient centrifugation and differential attachment.Immunofluorescence of neuron-specific enolase was adopted for the identification of the cultivated hippocampal neurons.Set 25 mmol/L glucose as the control group(CG)and 50 mmol/L glucose as the high glucose group(GG).The concentration of p38MAPK and JNK pathway inhibitor are both 10 μmol/L.CCK-8 was adopted to decide the optimum concentration of quercetin,oleanolic acid and icariin.Flow cytometry was used to detect the apoptosis of hippocampal neurons,and Western blot was adoted to detect the expressions of p-p38,p38,p-JNK and JNK.[Result]1.Hippocampal neurons were cultivated successfully,and the purity of them was 97.2±1.1%.2.CCK-8 was used to explore the optimum concentration and detect the vitality of neurons:compared with the CG,the relative activity of neurons in GG decreased(P<0.01);compared with the GG,the relative activity of neurons increased(P<0.01)when the concentration of quercetin,oleanolic acid and icariin were separately 10-5mol/L,10-6mol/L,and 10-5mol/L,which were determined as the best concentration.3.Flow cytometry apoptosis detection:Compared with the CG,the neuronal apoptosis rate of GG increased(P<0.01);Compared with the GG,the neuronal apoptosis rate of other groups were all decreased(P<0.01);There was no significant difference in the neuronal apoptosis rate between quercetin group(QG),oleanolic acid(NG),icariin group(YG),p38MAPK inhibitor group(p38 IG)and JNK inhibitor group(JNK IG).Respectively compared with the QG,NG and YG,the neuronal apoptosis rates of quercetin+oleanolic acid group(QN),quercetin+icariin group(QY),oleanolic acid+icariin group(NY)and quercetin+oleanolic+icariin group(QNY)were all decresed(P<0.01).Respectively compared with the QN,QY and NY,the neuronal apoptosis rates of QNY decresed(P<0.05).There was no significant difference in the neuronal apoptosis rate between QN,QY and NY.4.Western Blot:Compared with the CG,the p-p38/p38 of GG increased(P<0.01);Compared with the GG,the p-p38/p38s of QG,NG,YG,QN,QY,NY,QNY and p38 IG were all decreased(P<0.01),and the p-p38/p38 of JNK IG was increased(P<0.01);Compared with the p38 IG,the p-p38/p38s of QG,NG,YG and J-NK IG were all increased(P<0.01).Respectively compared with the QG,NG and YG,the p-p38/p38s of QN,QY,NY and QNY were decreased(P<0.01).There was no significant difference between QN,QY,NY and QNY.Compared with the CG,the p-JNK/JNK of GG increased(P<0.01);Compared with the GG,the p-JNK/JNKs of other groups were all decreased(P<0.01);Compared with the JNK IG,the p-JNK/JNKs of QG,NG,YG and p38 IG were all increased(P<0.01).Respectively compared with the QG,NG and YG,the p-JNK/JNKs of QN,QY,NY and QNY were decreased(P<0.01).Compared with the QN,the p-JNK/JNKs of QY and NY were increased(P<0.01),and the p-JNK/JNK of QNY was decreased(P<0.01);Compared with the QY,the p-JNK/JNKs of QY and NY were increased(P<0.01),and the p-JNK/JNK of QNY was decreased(P<0.01);Compared with the NY,the p-JNK/JNK of QNY was decreased(P<0.01).[Conclusion]1.High glucose promoted the apoptosis of hippocampal neurons,and the p-p38/p38 and p-JNK/JNK were both increased significantly,which indicated that the p38MAPK and J-NK signaling pathways were activated.2.Under the condition of high glucose,quercetin,oleanolic acid and icariin can alleviate the apoptosis of hippocampus neurons,reduce the phosphorylation of p38 and JNK in p38MAPK and JNK signaling pathway.3.Compared with the single drug,the compatibility between quercetin,oleanolic acid and icariin can further alleviate the apoptosis of hippocampus neurons cultured in high glucose,reduce the phosphorylation of p38 and JNK in p38MAPK and JNK signaling pathway.[Innovation]The compatibility of quercetin,oleanolic acid and icariin was used to interfere with hippocampal neurons cultured in high glucose,and the mechanism of apoptosis was studied from p38MAPK and JNK signaling pathways.