电针干预GDNF基因修饰神经干细胞对大鼠脊髓损伤后Caspase-8表达的影响OA

Objective:To explore methods for in vitro isolation and culture of high-purity neural stem cells(NSCs)and their biological characteristics,and to evaluate intervention effects of electroacupuncture combined with glial cell line-derived neurotrophic factor(GDNF)gene-modified NSC transplantation on cell apoptosis and motor function in rats after spinal cord injury.Methods:Fetal mouse brain tissues were isolated for in vitro NSC culture,and lentivirus carrying the GDNF gene was used for infection to assess transfection efficiency and cell growth status.T11 segment spinal cord incomplete transection model was established in 60 adult male SD rats,which were randomly di-vided into normal,model,NSC transplantation,and electroacupuncture intervention groups.BBB score was used to dynamically evaluate hindlimb motor function recovery.Samples were collected at different time points for assess-ment of Caspase-8 expression using immunohistochemistry and western blot.Results:A stable in vitro NSC culture system was successfully established,and primary cells showed good proliferation ability and multipotent differentia-tion potential.Behavioral evaluation showed that both NSC transplantation group and electroacupuncture intervention group had increased BBB scores compared with the model control group,and difference between the two groups was not statistically significant(P＞0.05).Apoptosis marker analysis showed that compared with the model control group,Caspase-8 expression levels were significantly downregulated in both the NSC transplantation group and the electroacupuncture intervention group(P＜0.05),with greater decrease in the electroacupuncture group(P＜0.05).Con-clusion:NSCs cultured in vitro can maintain stable proliferation and differentiation potential within passages 2-3.GDNF gene-modified NSC transplantation can effectively inhibit cell apoptosis after spinal cord injury.Electroacupuncture could further enhance anti-apoptotic effect,although no significant advantage in improving motor function has been observed yet.