珍宝丸联合电针对急性脊髓损伤大鼠行为学和BrdU、Nestin、GFAP表达的影响

Effects of Zhenbao Pills combined with electroacupuncture on ethology and expressions of BrdU,nestin and GFAP in rats with acute spinal cord injury

目的探讨珍宝丸联合电针对急性脊髓损伤大鼠行为学和BrdU、Nestin、GFAP表达的影响。方法建立急性脊髓损伤模型,SD成年大鼠随机分为正常对照组、模型组、丸剂组(0.54 g/kg)、电针组和联合组。丸剂组予以蒙药珍宝丸(0.54 g/kg)治疗,电针组予以电针干预,联合组予以蒙药珍宝丸联合电针治疗,共1周。以BBB量表评价治疗前后大鼠行为学变化,HE染色、TUNLE染色分别观察损伤脊髓组织的病理变化、细胞凋亡率,免疫荧光染色检测vWF阳性血管数目,Western blot、RT-PCR检测损伤脊髓组织BrdU、Nestin、GFAP蛋白及mRNA表达。结果治疗后,与正常对照组比较,模型组BBB评分下降(P<0.01),损伤脊髓组织分级升高(P<0.05),凋亡率增加(P<0.01),vWF阳性血管数目增多(P<0.01),BrdU、Nestin、GFAP蛋白及mRNA表达增加(P<0.01);与模型组比较,丸剂组、电针组和联合组BBB评分升高(P<0.01),损伤脊髓组织分级降低(P<0.05),凋亡率降低(P<0.01),vWF阳性血管数目增多(P<0.01),BrdU、Nestin、GFAP蛋白及mRNA表达增加(P<0.01),且联合组效果更佳。结论蒙药珍宝丸、电针治疗急性脊髓损伤大鼠均可改善其行为学、减轻病理变化、促进新生血管形成,二者联用的效果更佳,且推测很可能与上调损伤脊髓组织的BrdU、Nestin、GFAP表达有关。

AIM To explore the effects of Zhenbao Pills combined with electroacupuncture on ethology and expressions of BrdU,nestin and GFAP in rats with acute spinal cord injury.METHODS Adult SD rats were randomly divided into normal control group,model group,Zhenbao Pills group(0.54 g/kg),electroacupuncture group,and combination group with use of both Zhenbao Pills and electroacupuncture,and acute spinal cord injury model was established as well.Each group underwent one week corresponding administration.The rats were subjected to evaluation of their behavioral changes before and after treatment by BBB scale,the observation of pathological changes and apoptosis rate of spinal cord injury tissues by HE staining and TUNLE staining,the detection of number of vWF positive blood vessels by immunofluorescence staining,and determination of the protein and mRNA expressions of BrdU,Nestin and GFAP in spinal cord injury tissues by Western blot and RT-PCR.RESULTS After treatment,compared with the normal control group,the model group was observed with decreased BBB score(P<0.01),higher grade of injured spinal cord tissue(P<0.05),increased apoptosis rate(P<0.01),increased number of vWF positive blood vessels(P<0.01),increased BrdU,Nestin,GFAP protein and mRNA expression(P<0.01).Compared with the model group,Zhenbao Pills group,electroacupuncture group and the combinative intervention group shared increased BBB scores(P<0.01),lower grade of spinal cord injury tissue(P<0.05),decreased apoptosis rate(P<0.01),increased number of vWF positive blood vessels(P<0.01),increased protein and mRNA expressions of BrdU,Nestin,GFAP(P<0.01),and the combinative intervention group took the lead.CONCLUSION Upon rats with acute spinal cord injury,the combinative use of Zhenbao Pills and electroacupuncture can improve their ethology,and alleviate the pathological changes in terms of formation promotion of new blood vessels.It is suggested that this intervention may be associated with the up regulation of BrdU,nestin and GFAP expressions in spinal cord injury tissue.