牛膝杯苋甾酮激发大鼠骨髓间充质干细胞体外迁移及CXCR4表达的研究
Study on the in vitro migration and CXCR4 expression of rat bone marrow mesenchymal stem cells stimulated by cyasterone
  
DOI:10.3969/j.issn.1006.7108.2019.11.009
中文关键词:  杯苋甾酮  骨髓间充质干细胞  迁移  活血法  骨质疏松
英文关键词:cyasterone  bone marrow mesenchymal stem cells  migration  activating blood circulation method  osteoporosis
基金项目:国家自然科学基金(81503593,81473699);广东省自然科学基金(2016A030313649,2017A030313729);广东省省级科技技术项目(2014A020221055);广州中医药大学2017年“薪火计划”资助项目清单(XH20170104)
作者单位
陈鸿泰1,2 罗毅文2 陈东风1 徐亮亮1,3 刘亚梅1* 王斌2* 谢平金4 1.广州中医药大学广东 广州 510405 2.广州中医药大学第三附属医院广东 广州 510240 3.广州中医药大学第一附属医院广东 广州 510405 4.上海中医药大学深圳医院/深圳市罗湖区中医院广东 深圳518000 
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中文摘要:
      目的 研究牛膝杯苋甾酮激发大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)体外迁移及CXCR4蛋白表达的影响。方法 按照牛膝杯苋甾酮5、10、20 μg/mL浓度干预BMSCs,进行 MTT法、Transwell 迁移小室及PCR测试,确定杯苋甾酮激发BMSCs迁移的最佳浓度;通过siRNA沉默CXCR4,进一步验证SDF-1/CXCR4 信号轴在杯苋甾酮干预BMSCs迁移中的作用。结果 MTT法、Transwell 及PCR结果显示,以10 μg/mL浓度杯苋甾酮干预BMSCs迁移效果最好,且CXCR4表达量最高(P<0.05)。用siRNA沉默CXCR4后,4组细胞的迁移能力从低到高的顺序均为CXCR4沉默组、空白对照组、杯苋甾酮+CXCR4沉默组、杯苋甾酮组(P<0.05),与CXCR4蛋白表达一致。结论 活血药牛膝提取物杯苋甾酮能激发大鼠BMSCs 体外迁移,其相关机制可能与上调 CXCR4 蛋白表达有关。
英文摘要:
      Objective To study the effects of cyasterone on stimulating the in vitro migration of rat bone marrow mesenchymal stem cells (BMSCs) and the expression of CXCR4 protein. Methods BMSCs were intervened according to the concentration of 5, 10 and 20 μg/mL of cyasterone. The MTT assay, Transwell migration chamber and PCR test were performed to determine the optimal concentration of cyasterone to stimulate the migration of BMSCs. The siRNA was used to silence CXCR4 to further verify the role of the SDF-1/CXCR4 signal axis in the migration of BMSCs intervened using cyasterone. Results The results of MTT assay, Transwell and PCR showed that the migration of BMSCs with the concentration of 10 μg/mL cyasterone was the best, and the expression of CXCR4 was the highest (P<0.05). After silencing CXCR4 with siRNA, the order of migration ability of the four groups of cells from low to high was CXCR4 silencing group, blank control group, cyasterone + CXCR4 silencing group, and cyasterone group (P<0.05). It is consistent with the expression of CXCR4 protein. Conclusion Cyasterone, the extractive of radix cyathulae belong to blood-activating medicinal, could stimulate the migration of rat BMSCs in vitro. The related mechanism may be related to the up-regulation of CXCR4 protein expression.
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