The therapeutic effect and mechanism exploration of polydatin on acute spinal cord injury in mice
10.3969/j.issn.1004-406X.2025.09.05
- VernacularTitle:虎杖苷对急性脊髓损伤小鼠的治疗作用及机制探究
- Author:
Dan LUO
1
;
Sheng LUO
;
Yonghui HOU
Author Information
1. 广州中医药大学第二附属医院脊柱微创科 510000广州市;广东省中医药科学院 510000广州市
- Publication Type:Journal Article
- Keywords:
Polydatin;
Spinal cord injury;
Neurons;
Autophagy;
Apoptosis
- From:
Chinese Journal of Spine and Spinal Cord
2025;35(9):930-938
- CountryChina
- Language:Chinese
-
Abstract:
Objectives:To investigate the mechanism underlying the protective effects of polydatin(PD)a-gainst oxygen-glucose deprivation(OGD)-induced neuronal injury and acute spinal cord injury(SCI).Methods:The mouse hippocampal neuronal cell line HT22 was used in in vitro experiments.The cytotoxicity of PD was assessed using the CCK-8 assay:cells were treated with 0,1,3,10,30 and 100μM PD for 24h to de-termine the safe concentration range.After OGD modeling(24h),cells were treated with different concentrations of PD(0,3,10,and 30μM),and cell viability was measured via CCK-8 to identify the optimal protective concentration.Autophagy markers(LC3 and P62)were detected by immunofluorescence and Western blot(WB);Autophagosome formation was observed using transmission electron microscopy;And the apoptosis rate was e valuated by TUNEL staining.For in vivo studies,an acute SCI model was established in C57BL/6 mice using Allen's impact method.Animals were divided into sham,SCI model,and PD treatment(20mg/kg)groups,with n=10 per group.Tissues were collected on 7d and 14d post-injury.Spinal cord pathology was examined by HE staining on 7d and 14d.Immunofluorescence was performed on 14d to evaluate glial scar formation(GFAP+),neuronal survival(MAP2+),cell proliferation(BrdU+),and autophagy level(LC3 Ⅱ).Results:In vitro results showed that 1-100μM PD did not exhibit significant cytotoxicity toward HT22 cells.The optimal concentration of PD was determined to be 10μ M after OGD induction.Compared with the OGD group,10μM PD significantly enhanced cell viability after OGD[OGD group:(54.63±3.90)％vs OGD+PD(1 0μ M)group:(84.35±1.38)％,P＜0.05]and effectively attenuated autophagy activation,as evidenced by a decreased LC3Ⅱ/Ⅰratio(OGD group:11.0±0.57 vs OGD+PD group:3.50±0.28,P＜0.05),increased P62 accumulation(OGD group:0.55±0.04 vs OGD+PD group:0.93±0.06,P＜0.05),reduced number of autolysosomes,and significantly lower apoptosis rate[OGD group:(35.33±2.6)％vs OGD+PD group:(19.67±1.76)％,P＜0.05].In vivo,HE staining confirmed that pathological damage in spinal cord tissue was markedly alleviated in the PD-treated group compared with the SCI model group.Immunofluorescence results indicated that PD treatment inhibited fibrous scar formation(SCI group:2.32±0.19mm2 vs PD group:1.07±0.24mm2,P＜0.05),reduced neuronal damage(SCI group:1.72±0.28mm vs PD group:0.93±0.12mm,P＜0.05)and promoted cell proliferation[SCI group:(16.14±2.24)％vs PD group:(39.09±3.16)％,P＜0.05],and downregulated LC3 Ⅱ expression(SCI group:62.81±5.25 vs PD group:34.09±3.98,P＜0.05).Conclusions:PD ameliorates neural damage by concurrently suppressing autophagy and apoptosis,providing a dual-pathway therapeutic strategy for SCI.
