丹参酮ⅡA通过调控星形胶质细胞表型极化促进脊髓损伤修复  

TanshinoneⅡA promotes the repair of spinal cord injury by regulating astrocyte phenotype polarization

作　　者：饶江文 刘向春[1] 贺丰[1] 付玲玲 王小钢 刘宸 魏峰 马小芳 朱红霞 姜涛[1] RAO Jiangwen;LIU Xiangchun;HE Feng;FU Lingling;WANG Xiaogang;LIU Chen;WEI Feng;MA Xiaofang;ZHU Hongxia;JIANG Tao(Department of Orthopedics,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 101100,China;First Clinical Medical School,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学东直门医院骨科,北京101100 [2]北京中医药大学第一临床医学院,北京100700

出　　处：《贵州医科大学学报》2024年第11期1622-1630,1638,共10页Journal of Guizhou Medical University

基　　金：国家自然科学基金(82104900)。

摘　　要：目的观察丹参酮ⅡA(TⅡA)对脊髓损伤模型大鼠脊髓组织星形胶质细胞表型极化及运动功能的影响。方法将90只雌性SD大鼠随机均分为假手术组、模型组、TⅡA组、星形胶质细胞抑制剂组(LAA组)及TⅡA+星形胶质细胞抑制剂组(TⅡA+LAA组),假手术组仅咬除T8-T10椎板(不损伤硬脊膜),后4组大鼠为脊髓横断损伤模型大鼠,后3组大鼠损伤部位注射相应药物;造模成功后于术后第3、7、14、21、28、56天时分别进行行为学观察和巴索-比蒂-布雷斯纳汉运动功能评分(BBB评分),并取损伤段脊髓组织采用免疫组织荧光染色以及苏木精-伊红(HE)染色、尼氏染色观察脊髓组织学改变;采用qRT-PCR检测各组细胞中A1星形胶质细胞标志物C3、A2星形胶质细胞标志物S100A10、神经胶质细胞标志物GFAP、神经元标志物NF200的表达,Western blot检测各组细胞C3、S100A10、GFAP蛋白表达。结果与模型组比较,TⅡA组大鼠BBB评分升高,S100A10 mRNA和蛋白表达升高,NF200 mRNA和蛋白表达升高,GFAP mRNA和蛋白表达升高,C3 mRNA和蛋白表达降低,差异均有统计学意义(P<0.05);与模型组相比,TⅡA组尼氏体数目无明显差异;TⅡA可改善LAA造成的星形胶质细胞增殖抑制,激活反应性星形胶质细胞;与LAA组相比,TⅡA组S100A10蛋白表达升高(P<0.001);与LAA组相比,TⅡA+LAA组S100A10蛋白表达升高(P<0.0001)。结论TⅡA腹腔注射可改善SCI大鼠的运动功能,其机制可能与调控反应性星形胶质细胞表型,促进A2型星形胶质细胞极化以及减少脊髓病理损伤有关。Objective To observe the effect of tanshinoneⅡA(TⅡA)on astrocyte phenotype polarization in spinal cord tissue in a rat spinal cord injury(SCI)model and motor function.Methods Ninety female SD rats were randomly divided into sham group,model group,TⅡA group,astrocyte inhibitor group(LAA group)and TⅡA+astrocyte inhibitor group(TⅡA+LAA group).Sham group was bit off T8-T10 laminae(without damaging dura mater).The last 4 groups of rats were used to create a spinal cord transection injury model,and the last 3 groups of rats were injected with corresponding drugs at the site of injury.After successful modeling,behavioral observations and Basso-Beattie-Bresnahan(BBB)locomotor rating scale were performed on the 3 rd,7 th,14 th,21 th,28 th,and 56 th postoperative days,respectively.The injured spinal cord tissues were collected and subjected to immunohistochemical staining(IHC)and immunofluorescent staining(IF),hematoxylin&eosin(H&E)staining and Nissl staining to observe histological changes in the spinal cords.qRT-PCR was used to detect C3 expression of A1 astrocyte marker,S100A10 expression of A2 astrocyte marker,glial cell marker GFAP and neuronal marker NF200 in each group of cells.Western blot was used to detect the protein expression of C3,S100A10,and GFAP in each group of cells.Results When compared to model group,TⅡA group had increased BBB score,the elevated mRNA and protein expressions of S100A10,NF200 and GFAP as well as reduced C3 mRNA and protein expressions(P<0.05).There was no significant difference in Nissl body number between model and TⅡA groups.TⅡA improved LAA-mediated proliferation inhibition of astrocytes and activated reactive astrocytes.When compared to LAA group,TⅡA group had elevated S100A10 protein expression(P<0.001).When compared to LAA group,TⅡA+LAA group showed an increase in S100A10 protein expression(P<0.0001).Conclusion Intraperitoneal injection of TⅡA can improve the motor function of rats with SCI.Its mechanism may be related to regulating the phenotype of reactive a

关 键 词：脊髓损伤 丹参酮ⅡA 星形胶质细胞 A1星形胶质细胞 A2星形胶质细胞 

分 类 号：R681.5[医药卫生—骨科学] R361.2[医药卫生—外科学]