延龄草甾体皂苷对脑缺血大鼠星形胶质细胞GFAP、Cx43及Wnt3a/Dvl3信号通路的影响  

Effect of Total Steroid Saponin of Trillium tschonoskii Maxim.on Astrocyte GFAP,Cx43 and Wnt3a/Dvl3 Signaling Pathway in Rats with Cerebral Ischemia

作　　者：李慢中 欧阳俊摇 杨乐[2] 庄雨明 詹宇[2] 邹海艳[2] 赵晖[2] LI Manzhong;OUYANG Junyao;YANG Le;ZHUANG Yuming;ZHAN Yu;ZOU Haiyan;ZHAO Hui(Department of Pharmacy,Beijing Shijitan Hospital,Capital Medical University,Beijing 100038,China;School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China;Beijing Key Laboratory of Bio-characteristic Profiling for Evaluation of Rational Drug Use,Beijing 100038,China)

机构地区：[1]首都医科大学附属北京世纪坛医院药学部,北京100038 [2]首都医科大学中医药学院,北京100069 [3]临床合理用药评价北京市重点实验室,北京100038

出　　处：《中国中医药信息杂志》2022年第5期75-80,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金青年基金(82104974);北京市教育委员会科技发展计划面上项目(KM201510025011)。

摘　　要：目的观察延龄草甾体皂苷(TSTT)对脑缺血大鼠星形胶质细胞胶质纤维酸性蛋白(GFAP)、缝隙连接蛋白43(Cx43)及Wnt3a/Dvl3信号通路的影响,探讨其改善脑缺血的作用机制。方法采用线栓法制备局灶性脑缺血模型,将SD大鼠随机分为假手术组、模型组、金纳多组(60 mg/kg)及TSTT低、高剂量组(33、65 mg/kg),每组12只。分别进行相应干预,连续15 d。采用透射电镜观察神经元、星形胶质细胞超微结构,免疫荧光染色检测缺血边缘皮层、纹状体GFAP和Cx43表达,RT-PCR检测缺血周围脑组织Wnt3a、Dvl3基因表达。结果透射电镜观察结果显示:模型组大鼠神经元核明显固缩,核仁消失,染色质电子密度增加,细胞质内细胞器明显减少,可见大量空泡,星形胶质细胞肿胀明显,染色质消融,核周围出现空白区,细胞器减少;与模型组比较,TSTT低、高剂量组和金纳多组大鼠神经元和星形胶质细胞超微结构损伤明显减轻。免疫荧光染色结果显示:与假手术组比较,模型组大鼠缺血边缘皮层、纹状体GFAP和Cx43表达显著升高(P<0.01);与模型组比较,TSTT低、高剂量组和金纳多组大鼠缺血边缘皮层、纹状体GFAP和Cx43表达显著降低(P<0.01)。RT-PCR结果显示:与假手术组比较,模型组大鼠缺血周围脑组织Dvl3基因表达显著降低(P<0.01);与模型组比较,TSTT高剂量组和金纳多组大鼠缺血周围脑组织Wnt3a和Dvl3基因表达显著升高(P<0.05,P<0.01)。结论TSTT可通过上调Wnt3a/Dvl3基因表达、下调GFAP和Cx43表达,抑制星形胶质细胞异常活化,减轻脑缺血大鼠神经元、星形胶质细胞超微结构损伤。Objective To observe the effects of total steroid saponin of Trillium tschonoskii Maxim.(TSTT)on the expressions of astrocyte glial fibrillary acidic protein(GFAP)and Connexin-43(Cx43)and Wnt3a/Dvl3 signaling pathway in rats with cerebral ischemia;To explore its mechanism of improving cerebral ischemia.Methods SD rats were randomly divided into sham-operation group,model group,Ginaton group(60 mg/kg)and TSTT low-,high-dosage group(33,65 mg/kg),with 12 rats in each group.Rat model of middle cerebral artery occlusion was established with a suture method.The above groups received respectively intervention,for 15 days.The transmission electron microscope was used to detect the ultrastructural changes of the neurons and astrocytes.The immunofluorescence staining was conducted to observe the expressions of GFAP and Cx43 in the ischemic marginal cortex and striatum.RT-PCR was performed to detect the expression of Wnt3a,Dvl3 mRNA.Results The results of transmission electron microscopy showed that the nuclei of the neurons in the model group were obviously pyknotic,the nucleoli disappeared,the electron density of chromatin increased,the organelles in the cytoplasm were significantly reduced,a large number of vacuoles were seen,the astrocytes swelled significantly,the chromatin ablated,a blank area appeared around the the nucleus,and the organelles decreased.Compared with the model group,the ultrastructural damage of the rats’neurons and astrocytes in TSTT low-,high-dosage group and Ginaton group were significantly reduced.Immunofluorescence staining results showed that compared with the sham-operation group,the expressions of GFAP and Cx43 in the ischemic marginal cortex and striatum in the model group significantly increased(P<0.01).Compared with the model group,the expressions of GFAP and Cx43 in the ischemic marginal cortex and striatum in TSTT low-,high-dosage group and Ginaton group significantly decreased(P<0.01).RT-PCR results showed that compared with the sham-operation group,the expression of Dvl3 gene in brain

关 键 词：延龄草甾体皂苷 星形胶质细胞 GFAP CX43 Wnt3a/Dvl3信号通路 脑缺血 大鼠 

分 类 号：R285.5[医药卫生—中药学]