补阳还五汤联合脊髓间充质干细胞移植对大鼠脊髓外伤性截瘫后功能恢复的影响

Effect of Buyang Huanwu Decoction Combined with Bone Marrow Stromal Cells Transplantation on Functional Recovery after Spinal Cord Rraumatic Paraplegia in Rats

目的:探讨补阳还五汤联合脊髓间充质干细胞(BMSCs)移植对大鼠脊髓外伤性截瘫后功能恢复的影响,并初步探讨其可能的作用机制。方法:将大鼠分为5组,即假手术组、模型组、补阳还五汤组、BMSCs组、补阳还五汤+BMSCs组,各10只。采用改良Tarlov评分和斜板试验测定大鼠脊髓功能,HE染色观察脊髓损伤的情况,免疫组织化学染色检测脊髓组织中Brdu阳性细胞数;蛋白质印迹检测脊髓组织中gp130、IL-6蛋白表达。结果:在1、3、5周时,各治疗组Tarlov评分和倾斜平面临界角度明显高于模型组,gp130、IL-6蛋白表达明显低于模型组(P<0.05);补阳还五汤+BMSCs组Tarlov评分和倾斜平面临界角度明显高于BMSCs组,gp130、IL-6蛋白表达明显低于BMSCs组(P<0.05);移植5周后,补阳还五汤+BMSCs组Brdu标记阳性细胞数明显多于BMSCs组(P<0.05);HE染色见假手术组脊髓组织结构清晰,未见明显细胞结构破坏;模型组灰质破坏溶解,胶质疤痕形成,少见完整的细胞结构,较难区分灰、白质;各治疗组脊髓组织结构较清晰,灰质中坏死区较小,白质有存留,胶质疤痕少。结论:补阳还五汤联合BMSCs移植能促进大鼠脊髓外伤性截瘫后运动功能的恢复,并通过促进BMSCs迁移,抑制gp130、IL-6蛋白表达起作用。

Objective:To explore the effect of Buyang Huanwu Decoction combined with spinal cord mesenchymal stem cells(BMSCs)transplantation on functional recovery after spinal cord injury induced paralysis in rats,and to preliminarily explore its possible mechanism of action.Methods:Rats were divided into 5 groups,namely sham surgery group,model group,Buyang Huanwu Decoction group,BMSCs group,and Buyang Huanwu Decoction+BMSCs group,with 10 rats in each group.Modified Tarlov score and slant plate test were used to measure spinal cord function in rats,HE staining was used to observe spinal cord injury,and immunohistochemical staining was used to detect the number of Brdu positive cells in spinal cord tissue.Protein blotting was used to detect the expression of gp130 and IL-6 proteins in spinal cord tissue.Results:At weeks 1,3,and 5,the Tarlov score and critical angle of tilt plane in each treatment group were significantly higher than those in the model group,while the expression of gp130 and IL-6 proteins was significantly lower than that in the model group(P<0.05).The Tarlov score and critical angle of tilt plane in the Buyang Huanwu Decoction+BMSCs group were significantly higher than those in the BMSCs group,while the expression of gp130 and IL-6 proteins was significantly lower than those in the BMSCs group(P<0.05).After 5 weeks of transplantation,the number of Brdu labeled positive cells in the Buyang Huanwu Decoction+BMSCs group was significantly higher than that in the BMSCs group(P<0.05).HE staining showed that the spinal cord tissue structure of the sham operated group was clear,and no significant cellular structural damage was observed.In the model group,gray matter was destroyed and dissolved,glial scars formed,and complete cellular structures were rare,making it difficult to distinguish between gray and white matter.The spinal cord tissue structure of each treatment group is relatively clear,with small necrotic areas in the gray matter,preserved white matter,and fewer glial scars.Conclusion:Buyang Huanwu Decoction combined with BMSCs transplantation can promote the recovery of motor function in rats with spinal cord injury paraplegia,and inhibit the expression of gp130 and IL-6 proteins by promoting the migration of BMSCs.