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 Ttransplantation 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 investigate the effect of Buyang Huanwu Decoction Combined with BMSCs transplantation on the functional recovery of spinal cord traumatic paraplegia in Sprague-Dawley rats,and to explore its possible mechanism.Methods:The rats were divided into 5 groups,namely the sham group,model group,Buyang Huanwu decoction group,BMSCs transplantation group,Buyang Huanwu decoction+BMSCs group,10 rats in each group.Modified Tarlov score and inclined plane test were used to determine the spinal cord function of rats.HE staining was used to observe the spinal cord injury.Detect the number of Brdu-positive cells in spinal cord tissue(immunohistochemical staining).Western blotting was used to detect gp130,IL-6 protein expression.Results:At 1w,3w,and 5w,there were statistically significant differences in Tarlov score,critical angle of inclined plane and related protein expression between the sham operation group,each treatment group and the blank control group(P<0.05).At 1w,3w,and 5w,the Tarlov score and the critical angle of the inclined plane in the Buyang Huanwu decoction+BMSCs group were significantly higher than those in the MSCs group(P<0.05),and the expression of gp130,IL-6 protein was significantly lower than that in the BMSCs group(P<0.05).There were no Brdu-labeled positive cells in the spinal cord tissue of the sham-operated group.Brdu-positive BMSCs with brownish-yellow cells in the other BMSCs transplantation groups survived in the spinal cord injury area and surrounding tissues.BMSCs migrated and survived more 1 week after transplantation,and 5 weeks after transplantation.The number of Brub marker positive cells in Buyang Huanwu Decoction+BMSCs group was significantly higher than that in BMSCs group(P<0.05).HE staining showed clear spinal cord tissue structure and no obvious cell structure damage in the sham operation group.In the model group,gray matter was destroyed and dissolved,glial scar was formed,and the complete cell structure was rarely seen,and it was difficult to distinguish gray matter from white matter.In the treatment group,the spinal cord tissue structure was clear,the necrotic area in gray matter was small,the white matter remained,and the glial scar was less.Conclusion:Buyanghuwu Decoction combined with BMSCs transplantation can promote the recovery of motor function after spinal cord injury paraplegia in rats,and play a role by promoting BMSCs migration and inhibiting gp130 and IL-6 protein expression.