Preliminary Study On NMDA Receptors-Targeted Therapeutic Strategies For Ischemic Stroke

Cerebral ischemia is a common central nervous system disease characterized by high disability rate,high mortality,and high recurrence rate.In the modern era when a “super-aging” process occurs,cerebral ischemia has brought a huge economic burden to the society and families.In this context,a new administration strategy for patients with cerebral ischemia is essential.As NMDA receptor is an effective therapeutic target for cerebral ischemia and GluN2 A is one of the major subunits of NMDA receptor,this project focuses on GluN2 A subunit and provides a preliminary discussion on the feasibility of increased GluN2 A subunit function in the treatment of cerebral ischemia during the convalescent period,through using TCN-201(a GluN2 A antagonist)and curcumin that up-regulates the expression of GluN2 A subunit.Firstly,a methodology for long-term culture of high-purity hippocampal neurons was established by exploring how to realize the long-term culture of hippocampal neurons in vitro,and Western blotting was used to detect the expression of GluN2 A subunit in hippocampal neurons;secondly,the NMDA-induced excitotoxicity model was built based on the NMDA concentration and exposed time;finally,the curcumin-HP-β-CD inclusion complex was prepared,and evaluated for quality.The effects of TCN-201 and curcumin inclusion complex on NMDA-induced neuronal injury were assessed using a delayed administration regimen.The results showed that high-purity hippocampal neurons were successfully cultured under the conditions of the project,and GluN2 A increased with the culture time and reached a higher expression level at 11 Days;the NMDA-induced excitotoxicity model was preferably established at the NMDA concentration of 50 μM and incubated time of 20 min,for which a moderate injury can be developed;the curcumin-HP-β-CD inclusion complex was prepared from curcumin and HP-β-CD at the molar ratio for 1:1,during which the reaction temperature was 50°C,the stirring rate was 600 rpm,and the reaction time was 4 h;and NMDA-induced excitotoxicity would be aggravated if TCN-201 was administered after NMDA withdrawal,but would be the significantly inhibited if curcumin inclusion complex was given.As evidenced by the above results,the GluN2 A subunit may mediate pro-survival effects in the convalescent period of cerebral ischemia.Hence,the increases of GluN2 A function in the convalescent period of cerebral ischemia may present an effective administration strategy that is worthy of further study.