补阳还五汤通过调控PI3K/Akt、JAK2/STAT3信号促进BMSC趋化迁移对外伤性脊髓损伤大鼠神经元活性及认知功能的影响  

Buyang Huanwu Decoction promotes chemotaxis and migration of BMSC by regulating PI3K/Akt and JAK2/STAT3 signals and the effect on neuronal activity and cognitive function in rats with traumatic spinal cord injury

作　　者：宋颖军[1] 李旭[1] 刘小舟 张国福[1] SONG Ying-Jun;LI Xu;LIU Xiao-Zhou(Affiliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330006,Jiangxi,China)

机构地区：[1]江西中医药大学附属医院,江西南昌330006

出　　处：《中国老年学杂志》2023年第17期4206-4213,共8页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(81860856)。

摘　　要：目的研究补阳还五汤通过调控磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/Akt)、内源性酪氨酸激酶(JAK)2/信号传导和转录启动因子(STAT)3信号促进骨髓间充质干细胞(BMSCs)趋化迁移对外伤性脊髓损伤大鼠的神经元活性及认知功能的影响。方法选取健康大鼠53只,随机分为健康组(健康大鼠常规饲养)、损伤组(建立脊髓损伤模型)、干预组(补阳还五汤治疗)、对照组(甲泼尼龙治疗),每组12只,剩余5只大鼠用于补阳还五汤含药血清制备。流式细胞术鉴定BMSCs细胞。Transwell小室法测大鼠BMSCs迁移。高架十字迷宫和Morris水迷宫实验检测大鼠认知功能。苏木素-伊红(HE)染色检测脊髓组织病理形态。TUNEL测脊髓组织神经细胞凋亡。免疫组化检测p-JAK2、p-STAT3。Western印迹测PI3K、p-PI3K、Akt、p-Akt。结果传代后的培养细胞呈旋窝状或放射状贴壁生长,细胞多呈星形、梭形或三角状,培养3代后,细胞贴壁加快、形态均一,呈旋窝状或单层放射状生长。培养细胞表面抗原CD29、CD90为阳性,CD31、CD45为阴性,提示其为BMSCs细胞。与健康组相比,损伤组总路程、进入开臂次数、穿越平台次数显著降低,不同时间的潜伏期显著升高(P<0.05)。与损伤组相比,干预组与对照组总路程、进入开臂次数、穿越平台次数显著升高,不同时间的潜伏期显著降低(P<0.05)。干预组与对照组各指标对比无统计学差异(P>0.05)。健康组脊髓组织结构完整。损伤组脊髓组织疏松水肿,有细胞空泡变性产生。相较于损伤组,干预组与对照组大鼠脊髓组织病理形态有所改善。与健康组相比,损伤组BMSCs、PI3K、Akt、p-PI3K、p-Akt显著降低,神经细胞凋亡率、p-JAK2、p-STAT3显著升高(P<0.05)。与损伤组相比,干预组BMSCs、PI3K、Akt、p-PI3K、p-Akt显著升高,神经细胞凋亡率、p-JAK2、p-STAT3显著降低(P<0.05)。干预组与对照组各指标水平无统计学差异(P>0.05)。结论�Objective To study the effect of Buyang Huanwu Decoction on the neuronal activity and cognitive function of rats with traumatic spinal cord injury by regulating phosphatidyl inositol 3 kinase/protein kinase B(PI3K/AKT)and endogenous tyrosine kinase(JAK)2/signal transduction and activator of transcription(STAT)3 signals to promote the chemotaxis and migration of bone marrow mesenchymal stem cells(BMSCs).Methods Fifty-three healthy rats were selected and randomly divided into healthy group(conventional feeding of healthy rats),injury group(establishing a spinal cord injury model),intervention group(treatment with Buyang Huanwu Decoction)and control group(methylprednisone treatment),with 12 rats in each group,and the remaining 5 rats were used for the preparation of Buyang Huanwu Decoction-containing serum.BMSCs were identified by flow cytometry.Transwell cell method was used to measure the migration of rat BMSCs.The elevated plus maze and Morris water maze experiments were used to detect the cognitive function of rats.Pathological morphology of spinal cord tissue was detected by hematoxylin-eosin(HE)staining.TUNEL was used to measure the apoptosis of nerve cells in spinal cord tissue.Immunohistochemical was used to detect p-JAK2 and p-STAT3.PI3K,p-PI3K,Akt and p-Akt were measured by Western blotting.Results After passage,the cultured cells grew in a spiral or radial shape.The cells were mostly star-shaped,spindle-shaped or triangular.After 3 generations,the cells adhered to the wall faster and had uniform morphology,showing a spiral or single-layer radial shape grow.The cultured cell surface antigens CD29 and CD90 were positive,and CD31 and CD45 were negative,indicating that they were BMSCs cells.Compared with the healthy group,the total distance,the number of times to enter the open arms,the number of times to cross the platform were significantly reduced,and the incubation period of different time were significantly increased in injury group(P<0.05).Compared with the injury group,the total distance,the number of

关 键 词：补阳还五汤 磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/Akt) 内源性酪氨酸激酶(JAK)2/信号传导和转录启动因子(STAT)3 骨髓间充质干细胞(BMSCs)趋化迁移 神经元活性 认知功能 

分 类 号：R651.21[医药卫生—外科学]