Effect of Wendantang-containing Serum on Apoptosis and Expressions of PI3K，Akt and GSK3β in Astrocytes Under Glutamate Environment / 中国实验方剂学杂志

ABSTRACT

Objective:To investigate the protective effect of Wendantang-containing serum on astrocytes in glutamate environment and its effect on phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt)/glycogen synthetase kinase 3β (GSK3β) signal pathway. Method:Totally 60 rats were randomly divided into 5 groups, normal group (n=20), clozapine group, and high, medium and low-dose Wendantang groups, with 10 rats in each group. Normal group was given 20 mL·kg-1 normal saline, clozapine group was given 20 mg·kg-1 clozapine, Wendantang groups were given 40, 20, 10 g·kg-1 Wendantang, once a day. Eight days later, the rats were killed, their blood was taken, serum was centrifuged, inactivated, filtrated, sterilized and filled in separate centrifugal tubes. The astrocytes were divided into normal group, model group, clozapine group, and high, medium and low-dose Wendantang groups. The normal group and the model group were cultured with normal serum, and the rest groups were cultured with corresponding drug containing serum. The other groups were treated with 10 mmol·L-1 glutamic acid for 24 hours, and then the apoptosis of astrocytes was detected by flow cytometry. Western blot and Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) were used to determine protein, and mRNA expressions of PI3K, Akt, GSK3β in astrocytes. Result:Compared with the normal group, apoptosis in model group increased significantly (P<0.01). Compared with the model group, apoptosis in Wendantang groups decreased significantly (P<0.01). Compared with the normal group, protein and phosphorylation expressions of PI3K, Akt, GSK3β in model groups decreased significantly (P<0.05, P<0.01). Compared with the model group, protein and phosphorylation expressions of PI3K, Akt, GSK3β in Wendantang groups increased (P<0.05, P<0.01). Compared with the normal group, expressions of PI3K, Akt and GSK3β mRNA decreased in model group(P<0.01). Compared with the model group, expressions of PI3K, Akt and GSK3β mRNA increased significantly in Wendantang groups group (P<0.05，P<0.01). Conclusion:Wendantang-containing serum can effectively increase expressions of PI3K, Akt and GSK3β, so as to regulate PI3K/Akt/GSK3β signal pathway and protect nerve cells.