miR-141对股骨头坏死骨髓间质干细胞活性及VEGF/TGF-β<sub>2</sub>蛋白的影响

doi:10.3969/j.issn.1006-6233.2022.12.003

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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-β₂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-β₂蛋白及mRNA水平;荧光酶素实验证明miR-141对VEGF、TGF-β₂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-β₂表达降低(P<0.05),B组和C组VEGF、TGF-β₂表达无意义(P>0.05),与C组相比,D组VEGF、TGF-β₂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-β₂是miR-141靶基因,当抑制miR-141时VEGF、TGF-β₂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-β₂活性相关。

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	田楠楠
	冯蜜亚
	吴涛

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-β₂

Abstract：Objective: To investigate the effects of Mir-141 on the activity of bone marrow mesenchymal stem cells and VEGF/TGF-β₂ 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-β₂ were detected by western blot and RT-PCR, respectively. Luciferase assay demonstrated the regulatory effect of Mir-141 on VEGF and TGF-β₂. 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-β₂ in group B were decreased (P<0. 05), and the expressions of VEGF and TGF-β₂ in groups B and C were insignificant (P>0. 05). Compared with group C, the expressions of VEGF and TGF-β₂ in group D were increased, and there were differences among groups (P<0. 05). Luciferase assay confirmed that VEGF and TGF-β₂ were the target genes of Mir-141, and the expressions of VEGF and TGF-β₂ 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 head necrosis and inhibiting apoptosis. The mechanism may be related to the activation of VEGF and TGF-β₂ activities.

Key words： Necrosis of the femoral head Bone marrow mesenchymal stem cells miR-141 Vascular endothelial growth factor Transforming growth factor-β₂

基金资助:河北省医学科学研究基金项目,(编号:20201010)

通讯作者: 冯蜜亚

引用本文:

田楠楠, 冯蜜亚, 吴涛. miR-141对股骨头坏死骨髓间质干细胞活性及VEGF/TGF-β₂蛋白的影响[J]. 河北医学, 2022, 28(12): 1949-1955.
TIAN Nannan, FENG Miya, et al. Effects of miR-141 on Activity of Bone Marrow Mesenchymal Stem Cells and VEGF/TGF-β₂ Protein in Femoral Head Necrosis. HeBei Med, 2022, 28(12): 1949-1955.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2022.12.003 或 http://www.hbyxzzs.cn/CN/Y2022/V28/I12/1949

[1] Chan M J,Huang C C,Hu C C,et al.Osteonecrosis of femoral head,an overlooked long-term complication after paraquat Intoxication:a retrospective cohort study[J].Scientific Reports,2020,10(1):8827.
[2] He M C,J Zhang,XJ Chen,et al.Osteoclastic activity was associated with the development of steroid-induced osteonecrosis of femoral head[J].Artificial Cells,2020,48(1):1036-1046.
[3] Wu Y,Tang Y,Zhang X,et al.MMP-1 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells via the JNK and ERK pathway[J].The International Journal of Biochemistry & Cell Biology,2020,129(2):105880.
[4] 莫峰波.5'-氮杂胞苷对激素性股骨头坏死骨髓间充质干细胞分化作用的研究[D].华中科技大学,2014.
[5] Aanhold C,Bus P,Zandbergen M,et al.The Vascular Endothelial Growth Factor Inhibitor Soluble FLT-1 Ameliorates Atopic Dermatitis in APOC1 Transgenic Mice - ScienceDirect[J].Journal of Investigative Dermatology,2020,140( 2):491-494.
[6] 常晓朋,陈涛,赵寅,等.骨形态发生蛋白2和转化生长因子β₂协同促进骨髓间充质干细胞成骨分化[J].中国组织工程研究,2019,23(1):7-12.
[7] 孟晨阳,冯卫,薛飞,等.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[J].中国组织工程研究,2020,24(19):2991-2996.
[8] Wang M L,Nesti L J,Tuli R,et al.Titanium particles suppress expression of osteoblastic phenotype in human mesenchymal stem cells[J].Journal of Orthopaedic Research,2010,20(6):1175-1184.
[9] 孔令驰,孙兴亮,左荣台,等.早期移植生长分化因子11表达沉默的骨髓基质干细胞促进大鼠激素性股骨头坏死区成骨的研究[J].中华创伤骨科杂志,2019,21(9):802-809.
[10] Jianxin S,Gang W,Chunxiao L,et al.Screening for differentially expressed miRNAs in aedes albopictus (Diptera:Culicidae) exposed to DENV-2 and their effect on replication of DENV-2 in C6/36 cells[J].Parasites & vectors,2020,12(1):44.
[11] Qing,Cai,Peng,et al.MicroRNA‐224 enhances the osteoblastic differentiation of hMSCs via Rac1[J].Cell Biochemistry and Function,2019,37(2):62-71.
[12] Yu K R,Lee S,Jung J W,et al.MicroRNA-141-3p plays a role in human mesenchymal stem cell aging by directly targeting ZMPSTE24[J].Journal of Cell Science,2013,126(23):5422-5431.
[13] 张文明.miR-141调控OP小鼠BMSCs成骨分化机制及健骨颗粒干预作用研究[D].福建中医药大学,2016.
[14] 张志恒,张文明,魏振朴,等.健骨颗粒对去卵巢小鼠miR-141及成骨基因Dlx5/Msx2/Runx2的影响[J].康复学报,2018,30(6):26-31.
[15] 张楚天,张文明,林燕萍,等.健骨颗粒含药血清调控miR-141对小鼠骨髓间充质干细胞成骨分化的影响[J].中国骨质疏松杂志,2020,26(4):35-39+122.
[16] Choi S K,Kim H S,Jin T,et al.Overexpression of the miR-141/200c cluster promotes the migratory and invasive ability of triple-negative breast cancer cells through the activation of the FAK and PI3K/AKT signaling pathways by secreting VEGF-A[J].Bmc Cancer,2016,16(1):570.