摘要
目的研究中药红花有效成分棕榈酸是否通过靶向调控C-X-C趋化因子受体4促进大鼠骨髓间充质干细胞(rBMSCs)体外迁移。方法通过全骨髓贴壁培养法培养rBMSCs,取第三代(P3)rBMSCs进行实验。棕榈酸按照低、中、高浓度梯度(5g/L、10g/L、20g/L)干预处理rBMSCs,并进行Transwell实验,确定最佳促迁移浓度为10g/L。应用RT-qPCR检测不同浓度的棕榈酸对CXCR4 mRNA表达的影响,同时运用CXCR4 siRNA质粒转染rBMSCs,并分为棕榈酸组(10g/L)、CXCR4 siRNA组、棕榈酸(10g/L)+CXCR4 siRNA组及空白对照组(10g/L),进一步通过Transwell实验、RT-qPCR实验、Western Blot实验,验证棕榈酸靶向调控C-X-C趋化因子受体4(CXCR4)对干细胞迁移的影响。结果不同浓度的棕榈酸均可促进rBMSCs的迁移并上调rBMSCs的CXCR4 mRNA的表达,其中以中浓度10g/L最佳(P<0.05)。转染CXCR4 siRNA后rBMSCs的Transwell实验表明,棕榈酸促进rBMSCs迁移的能力与提高C-X-C趋化因子受体4表达相关。结论棕榈酸通过上调C-X-C趋化因子受体4的表达促进大鼠骨髓间充质干细胞体外迁移。
Objective To investigate whether palmitic acid,the active component of safflower,can target and regulate CXCR4 to promote the migration of rBMSCs in vitro.Methods rBMSCs were cultured by whole bone marrow adherent culture,and the third generation(P3)rBMSCs were taken for experiment.Transwell assay was performed on rBMSCs treated with palmitic acid according to low-medium and high-concentration gradient intervention with 5 g/L,10 g/L,and 20 g/L,and the optimal migration concentration was determined to be 10 g/L.Application RT-qPCR experiment testing the influence of different concentrations of palmitic acid for CXCR4 mRNA expression.What’s more,using CXCR4 siRNA plasmid transfection rBMSCs,and divided into palmitic acid group(10 g/L),CXCR4 siRNA group,CXCR4 siRNA+palmitic acid(10 g/L)group and NC group,further by Transwell experiment,RT-qPCR,Western Blot experiments palmitic acid targeted regulation of C-X-C chemokine receptor 4(CXCR4)in stem cell migration.Results Palmitic acid at different concentrations can promote the migration of rBMSCs and up-regulate the expression of CXCR4 mRNA in rBMSCs,with the best concentration at 10 g/L(P<0.05).Transwell assay of rBMSCs transfected with CXCR4 siRNA showed that palmitic acid has the ability to promote the migration of rBMSCs,which is related to c-x-c chemokine receptor 4.Conclusion The mechanism of palmitic acid promoting the migration of BMSCs in vitro is mainly related to up-regulating the expression of c-x-c chemokine receptor 4.
作者
朱俊朗
陈鸿泰
罗毅文
陈东风
徐亮亮
王斌
刘亚梅
ZHU Jun-lang;CHEN Hong-tai;LUO Yi-wen;CHEN Dong-feng;XU Liang-liang;WANG Bin;LIU Ya-mei(Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou Guandong,510240,China;School of Basic Medicine of Guangzhou University of Chinese Medicine y Guangzhou Guangdong 510063,China;First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou Guangdong 510405,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2020年第3期562-566,共5页
Lishizhen Medicine and Materia Medica Research
基金
广东省自然科学基金(2016A030313649
2017A030313729)
国家自然科学基金(81503593,81873326)
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(XH20170104)。
作者简介
朱俊朗(1994-),男(汉族),广东广州人,广州中医药大学第三附属医院2017级硕士研究生,主要从事中医骨伤科临床工作。;通讯作者:王斌(1973-),男(汉族),安徽天长人,广州中医药大学第三附属医院主任中医师,博士学位,主要从事中医创伤骨科临床和间充质干细胞生物学研究工作。
