To Investigate the Mechanism of Du Meridian Electroacupuncture Combined with Dual-Gene Modified Neural Stem Cell Transplantation in Rats with Spinal Cord Injury via the JAK2/STAT3 Pathway

YONG Chen; TAN Longwang; LI Jiang; CHEN Liheng; LIANG Jingliang; ZHANG Jiaxiang

Abstract：Objective To investigate the therapeutic effects and underlying mechanisms of Du meridian electroacupuncture combined with transplantation of dual-gene modified neural stem cells(NSCs)in rats with spinal cord injury(SCI).Methods Forty-eight SD rats were randomly divided into a blank group,a model group,a stem cell group,and a combination group,with 12 rats in each group.Except for the blank group,SCI models were established in the other groups using the modified Allen's weight-drop method.The stem cell group received an intramedullary injection of 5 μL(2×105 cells)of glial cell line-derived neurotrophic factor(GDNF)and hypoxia-inducible factor-1α(HIF-1α)dual-gene modified NSCs suspension at the injury site after modeling.The combination group received electroacupuncture at the Du meridian acupoints:"Dazhui(GV 14)","Zhiyang(GV 9)","Mingmen(GV 4)",and"Yaoyangguan(GV 3)"in addition to the stem cell treatment.General conditions of the rats were observed,and Basso,Beattie,and Bresnahan(BBB)locomotor scores were assessed at different time points.After sample collection,protein and mRNA levels related to SCI and NSCs differentiation,regeneration,and proliferation were measured.Bioinformatics enrichment analysis was performed on the study indicators,and the results were validated.Results GDNF and HIF-1α dual-gene modified NSCs were successfully cultured.After treatment,the combination group showed significant improvements in general conditions and BBB scores compared to the model group(P<0.01).Levels of inflammation,oxidative stress,and apoptosis in the injured tissue were significantly suppressed(P<0.01),and spinal cord pathology was improved.Protein expression of β-Catenin,Nestin,and heme oxygenase-1(HO-1)—factors associated with tissue repair—was significantly upregulated(P<0.01).Expression of glial fibrillary acidic protein(GFAP)decreased significantly,while β-III tubulin(Tuj-1)expression increased(P<0.01),indicating enhanced NSCs differentiation and regeneration.Bioinformatics enrichment analysis revealed that the targets involved were primarily enriched in the Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway.Animal experiments confirmed that JAK2/STAT3 pathway activation was significantly lower in the combination group than in the model group(P<0.05).Conclusion Electroacupuncture at the Du meridian combined with dual-gene modified NSCs transplantation can improve motor function and spinal cord pathology in SCI rats,promoting NSCs proliferation and differentiation.These effects may be associated with the inhibition of JAK2/STAT3 signaling pathway activation.