miR-141对股骨头坏死骨髓间质干细胞活性及VEGF/TGF-β_(2)蛋白的影响  

Effects of miR-141 on Activity of Bone Marrow Mesenchymal Stem Cells and VEGF/TGF-β_(2) Protein in Femoral Head Necrosis

作　　者：田楠楠 冯蜜亚 吴涛[2] TIAN Nannan;FENG Miya(Hengshui People's Hospital,Hebei Hengshui 053000,China)

机构地区：[1]河北省衡水市人民医院,河北衡水053000 [2]河北医科大学第三医院,河北石家庄050000

出　　处：《河北医学》2022年第12期1949-1956,共8页Hebei Medicine

基　　金：河北省医学科学研究基金项目,(编号:20201010)。

摘　　要：目的:探讨miR-141对股骨头坏死骨髓间质干细胞活性及VEGF/TGF-β_(2)蛋白的影响。方法:从我院收治的股骨骨折及激素性股骨头坏死患者手术期间抽取骨髓,培养及鉴定MSCs后随机分为四组,分别为A组(股骨颈骨折患者hMSCs细胞)、B组(激素性股骨头坏死患者hMSCs细胞)、C组(B组+miR-141-siRNA-NC)、D组(B组+miR-141-siRNA),MTT检测细胞活性;流式细胞仪检测凋亡率;免疫印迹及RT-PCR分别检测VEGF、TGF-β_(2)蛋白及mRNA水平;荧光酶素实验证明miR-141对VEGF、TGF-β_(2)调控作用。结果:与A组相比,B组hMSCs细胞24h、48h及72h活性降低(P<0.05),B组和C组24h、48h及72h hMSCs细胞OD值无意义(P>0.05),与C组相比,D组MSCs细胞24h、48h及72h活性升高,组间比较存在差异(P<0.05);A组、B组、C组及D组细胞凋亡率分别为(4.23±0.33)、(20.73±1.20)、(19.43±1.16)及(14.76±0.80),组间比较存在差异(F=381.00,P<0.001);与A组相比,B组VEGF、TGF-β_(2)表达降低(P<0.05),B组和C组VEGF、TGF-β_(2)表达无意义(P>0.05),与C组相比,D组VEGF、TGF-β_(2)表达升高,组间比较存在差异(P<0.05);荧光素酶实验证实VEGF、TGF-β_(2)是miR-141靶基因,当抑制miR-141时VEGF、TGF-β_(2)表达升高。结论:沉默miR-141能够通过提高股骨头坏死骨髓间质干细胞活性,抑制凋亡发挥髓间质干细胞保护作用,研究机制可能与激活VEGF、TGF-β_(2)活性相关。Objective:To investigate the effects of Mir-141 on the activity of bone marrow mesenchymal stem cells and VEGF/TGF-β_(2) protein in femoral head necrosis.Methods:Patients with femoral fracture and steroid-induced femoral head necrosis admitted to our hospital were randomly divided into 4 groups after bone marrow extraction,culture and identification of MSCs during surgery,including group A(hMSCs cells from patients with femoral neck fracture),group B(hMSCs cells from patients with steroid-induced femoral head necrosis),group C(group B+Mir-141-sirNA-NC),and group D(group B+Mir-141-SiRNA).MTT was used to detect cell activity.The apoptosis rate was detected by flow cytometry.The protein and mRNA levels of VEGF and TGF-β_(2) were detected by western blot and RT-PCR,respectively.Luciferase assay demonstrated the regulatory effect of Mir-141 on VEGF and TGF-β_(2).Results:Compared with group A,the activity of hMSCs cells in group B was decreased at 24h,48h and 72h(P<0.05),while the OD values of hMSCs cells in groups B and C were insignificant at 24h,48h and 72h(P>0.05).Compared with group C,the activity of MSCs cells in group D was increased at 24h,48h and 72h.There was significant difference between groups(P<0.05).The apoptosis rates of group A,B,C and D were(4.23±0.33),(20.73±1.20),(19.43±1.16)and(14.76±0.80),respectively,and there were differences among groups(F=381.00,P<0.001).Compared with group A,the expressions of VEGF and TGF-β_(2) in group B were decreased(P<0.05),and the expressions of VEGF and TGF-β_(2) in groups B and C were insignificant(P>0.05).Compared with group C,the expressions of VEGF and TGF-β_(2) in group D were increased,and there were differences among groups(P<0.05).Luciferase assay confirmed that VEGF and TGF-β_(2) were the target genes of Mir-141,and the expressions of VEGF and TGF-β_(2) increased when Mir-141 was inhibited.Conclusion:The silencing of Mir-141 can play a protective role of mesenchymal stem cells by improving the activity of bone marrow mesenchymal stem cells in femur

关 键 词：股骨头坏死 骨髓间质干细胞 miR-141 血管内皮生长因子 转化生长因子-β_(2) 

分 类 号：R681.8[医药卫生—骨科学]