Effects of astragaloside IV combined with Panax notoginseng saponins on proliferation, apoptosis, migration and neuronal differentiation of bone marrow mesenchymal stem cells in OGD/R model rats / 中草药

ABSTRACT

Objective: To observe the effects of astragaloside IV (AST IV) combined with Panax Notoginseng saponins (PNS) on proliferation, apoptosis, migration and neuronal differentiation of oxygen glucosedeprivation/reoxygenation model rat bone marrow mesenchymal stem cells (BMSCs). Methods: BMSCs were isolated, cultured, amplified and purified by the whole bone marrow adherent method. The positive expression rates of BMSCs surface markers, CD29, CD90, CD34, and CD45 were detected by flow cytometry. The third generation of BMSCs was pretreated with AST IV and PNS doses of high (100 μmol/L + 60 μmol/L), medium (50 μmol/L + 30 μmol/L), and low (25 μmol/L + 15 μmol/L) for 24 h. The model of ischemia-reperfusion injury was established by OGD/R. Meanwhile, the normal group (BMSCs were cultured normally) and the model group (OGD/R was used to establish an ischemia reperfusion injury model) were established. The cell increment rate was detected by CCK-8 assay. Cell apoptosis was detected by flow cytometry. Transwell assay was used to detect the migration of BMSCs. The condition of BMSCs differentiation into neurons and astrocytes was observed by Nestin/NSE and Nestin/GFAP immunofluorescence double labeling. Results: BMSCs were successfully cultured and separated, and the positive rates of CD29 and CD90 detected by flow cytometry were 94.23% and 94.69%, while the positive rates of CD34 and CD45 were 5.76% and 5.31%. Compared with the normal group, the survival rate of the model group was reduced significantly and the apoptosis rate was increased significantly (P < 0.05). Compared with the model group, the combination of different doses of AST IV and PNS could promote the proliferation of BMSCs (P < 0.05, 0.01) and inhibit the apoptosis (P < 0.05, 0.01). Compared with the normal group, the model group and the AST IV and PNS group at different doses could promote the migration of BMSCs (P < 0.05). Compared with the model group, the number of migrated cells in the AST IV and PNS groups at different doses was increased significantly (P < 0.05). Compared with the normal group, the model group and the AST IV and PNS groups at different doses could promote the differentiation of BMSCs into neurons and astrocytes (P < 0.01). Compared with the model group, the positive expression rates of Nestin/NSE and Nestin/GFAP in the AST IV and PNS groups at different doses were increased significantly (P < 0.01). Conclusion: AST IV combined with PNS can promote the proliferation and migration of BMSCs of ischemia-reperfusion model in vitro, inhibit the apoptosis, and induce their directional differentiation into neurons and astrocytes.