抑制芳香烃受体表达促进人脐带间充质干细胞的增殖和软骨分化  

Inhibition of aryl hydrocarbon receptor expression promotes the proliferation and chondrogenic differentiation of human umbilical cord mesenchymal stem cells

作　　者：王新伟 赵英杰[1] 李素素 王越业 贾成艳 常艳[1] 魏伟[1] Wang Xinwei;Zhao Yingjie;Li Susu(Institute of Clinical Pharmacology,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学临床药理研究所,合肥230032

出　　处：《安徽医科大学学报》2021年第9期1412-1417,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金面上项目(编号:81573443、81673444、81973332);安徽省自然科学杰出青年基金(编号:170808J10);高校优秀青年人才支持计划重点项目(编号:gxyqZD2016043)。

摘　　要：目的探究芳香烃受体(AhR)对人脐带间充质干细胞(hUC-MSCs)增殖和软骨分化能力的影响。方法用AhR-homo-1432 inhibitor和AhR-homo-1432 inhibitor阴性对照转染hUC-MSCs,设为si-AhR组和si-NC组。在转染hUC-MSCs后的第1、3、5、7天,CCK-8法测定细胞活力;软骨分化诱导培养hUC-MSCs 7 d,实时荧光定量PCR和Western blot检测软骨分化标志物SOX-9和COL2A1 mRNA和蛋白的表达;阿利新蓝染色检测蛋白聚糖的表达。结果与si-NC组比较,si-AhR组AhR的mRNA和蛋白水平表达降低,表明转染成功。CCK-8实验结果显示,与si-NC组比较,转染后的第5天和第7天si-AhR组hUC-MSCs活力增强。经软骨分化诱导培养后,与si-NC组比较,si-AhR组hUC-MSCs的SOX-9和COL2A1 mRNA和蛋白表达升高;阿利新蓝染色结果显示,si-AhR组hUC-MSCs表达的蛋白聚糖含量较si-NC组增多,染色更深,软骨分化能力增强。结论抑制AhR表达可促进hUC-MSCs的增殖和软骨分化。Objective To investigate the effect of aryl hydrocarbon receptor(AhR)on the proliferation and chondrogenic differentiation of human umbilical cord derived mesenchymal stem cells(hUC-MSCs).MethodsThe hUC-MSCs were transfected with AhR-homo-1432 inhibitor and AhR-homo-1432 inhibitor negative control,and set as si-NC group and si-AhR group.The proliferation ability of hUC-MSCs were measured by CCK-8 assay on day 1,3,5 and 7 after transfection.Real-time quantitative PCR and Western blot were used to detect the mRNA and protein expression of chondrogenic marker SOX-9 and COL2A1 after 7 days of the chondrogenic differentiation.Alcian blue staining was used to detect the expression of proteoglycan.Results Compared with si-NC group,the mRNA and protein expression of AhR in si-AhR group decreased which indicated the transfection of hUC-MSCs was successful.The results of CCK-8 showed that compared with si-NC group,the cell viability of si-AhR group improved on 5 and 7 day.After induction of chondrogenic differentiation,the results showed that compared with si-NC group,the expression of SOX-9 and COL2A1 in si-AhR group increased and Alcian blue staining showed that the si-AhR group had more proteoglycan contents,deeper staining and stronger cartilage differentiation ability.Conclusion Knocking down the expression of AhR can promote the proliferation and chondrogenic differentiation of hUC-MSCs.

关 键 词：人脐带间充质干细胞 芳香烃受体 细胞增殖 软骨分化 

分 类 号：R318[医药卫生—生物医学工程] R684.3[医药卫生—基础医学]