鹿茸血清调控miR-141影响地塞米松对骨髓间充质干细胞的促增殖作用  

Micro RNA-141 is regulated by serum of velvet antler to promote the proliferation of bone marrow mesenchymal stem cells in dexamethasone-induced cell model

作　　者：孟晨阳 薛飞[1] 贾燕飞[1] 佟雁翔[1] 张立峰[1] 于成涌[1] 张哲汉 王文选[1] 郝廷[1] 冯卫[1] Meng Chenyang;Xue Fei;Jia Yanfei;Tong Yanxiang;Zhang Lifeng;Yu Chengyong;Zhang Zhehan;Wang Wenxuan;Hao Ting;Feng Wei(Department of Orthopedics,Second Affiliated Hospital of Inner Mongolia Autonomous Region,Hohhot 010030,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古医科大学第二附属医院骨科

出　　处：《中国组织工程研究》2020年第19期2991-2996,共6页Chinese Journal of Tissue Engineering Research

基　　金：内蒙古自治区卫生和计划生育委员会科研计划项目(201703122)，项目负责人：冯卫~~

摘　　要：背景:骨髓间充质干细胞的增殖与成骨分化可延缓激素性股骨头坏死的进程,miR-141是促进细胞增殖的重要调控因子之一,鹿茸作为传统中药,在骨与组织修复及改善健康方面有显著的作用。目的:探讨鹿茸血清是否具有调控miR-141的作用,从而影响骨髓间充质干细胞的增殖,进而延缓或逆转激素性骨坏死的病程。方法:分离培养SD大鼠骨髓间充质干细胞,将第3代骨髓间充质干细胞行miRNA-141 mimic和miRNA-141 inhibitor转染,采用RT-PCR检测miR-141的mRNA表达,MTT比色法检测骨髓间充质干细胞的增殖能力。将第3代骨髓间充质干细胞分为3组:正常组、地塞米松组、鹿茸血清组,正常组用α-MEM培养基培养,地塞米松组在α-MEM培养基中加入终浓度在1μmol/L的地塞米松,鹿茸血清组在α-MEM培养基中加入终浓度在1μmol/L的地塞米松和体积分数为15%的鹿茸血清,干预24 h后采用RT-PCR检测miR-141的mRNA表达,干预24,48,72 h采用MTT比色法检测细胞增殖能力。结果与结论:①转染miR-141 mimic质粒后骨髓间充质干细胞中miR-141 mRNA表达升高,细胞增殖能力下降;转染miR-141 inhibitor质粒后骨髓间充质干细胞中miR-141 mRNA表达降低,细胞增殖能力升高;②地塞米松组miR-141的mRNA表达较正常组显著增加(P<0.01),鹿茸血清组miR-141的mRNA表达较地塞米松组显著下调(P<0.01);③地塞米松组骨髓间充质干细胞吸光度值低于正常组(P<0.01),鹿茸血清组吸光度值均高于地塞米松组(P<0.01);④由此可见,鹿茸血清可以抑制地塞米松引起的miR-141高表达,从而促进骨髓间充质干细胞增殖。BACKGROUND:The proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)can delay the procession of steroid-induced femoral head necrosis.Besides,microRNA-141(miR-141)is one of the important regulatory factors to promote cell proliferation.In addition,velvet antler is a traditional Chinese medicine which has significant roles in repairing bone and tissue and improving health.OBJECTIVE:To investigate whether velvet antler serum can regulate the expression of miR-141 to promote the proliferation of BMSCs,and further delay or reverse the progression of steroid-induced femoral head necrosis.METHODS:BMSCs were isolated and cultured from Sprague-Dawley rats.The passage 3 BMSCs were transfected with miR-141 mimic or miR-141 inhibitors,and then real-time PCR and methyl thiazolyl tetrazolium(MTT)assay were performed for detecting miR-141 expression and cell proliferation,respectively.The passage 3 BMSCs were divided into three groups:control group(α-MEM),dexamethasone group(α-MEM+1μmol/L dexamethasone),and velvet antler serum group(α-MEM+1μmol/L dexamethasone+15%velvet antler serum).Expression of miR-141 mRNA was detected by real-time PCR at 24 hours after intervention.The proliferation ability of BMSCs was evaluated by MTT assay at 24,48,and 72 hours after intervention.RESULTS AND CONCLUSION:After transfection with miR-141 mimic,the expression of miR-141 mRNA was upregulated,while the cell proliferation was reduced.After transfection with miR-141 inhibitor,the expression of miR-141 mRNA was downregulated,while the cell proliferation was increased.The expression of miR-141 mRNA was significantly higher in the dexamethasone group than the control group(P<0.01),while the treatment with velvet antler serum could significantly downregulate the expression of miR-141 mRNA(P<0.01).The absorbance of BMSCs in the dexamethasone group was significantly lower than that in the control group(P<0.01),and the absorbance value in the velvet antler serum group was significantly higher than that in the d

关 键 词：骨组织工程 激素性骨坏死 地塞米松 微小RNA-141 骨髓间充质干细胞 鹿茸 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]