| 马文贤,黄琼,董振耀.微小 RNA-218-5p靶向 Jagged 1调控人牙周膜干细胞的活性和骨向分化[J].安徽医药,2021,25(12):2503-2508. |
| 微小 RNA-218-5p靶向 Jagged 1调控人牙周膜干细胞的活性和骨向分化 |
| MicroRNA-218-5p regulates the activity and osteogenic differentiation of human periodontal ligament stem cells by targeting Jagged 1 |
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| DOI：10.3969/j.issn.1009-6469.2021.12.040 |
| 中文关键词: 微小 RNA-218-5p Jagged 1 牙周韧带 人牙周膜干细胞 骨向分化 |
| 英文关键词: MicroRNA-218-5p Jagged 1 Periodontal ligament Human periodontal ligament stem cells Osteogenic differen. |
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| 中文摘要: |
| 目的研究微小 RNA-218-5p(miR-218-5p)对人牙周膜干细胞( hPDLSCs)活性和骨向分化的影响并探讨其机制，为牙周炎的治疗提供研究基础。方法将 anti-miR-218-5p组(转染 anti-miR-218-5p)、 anti-miR-NC组(转染 anti-miR-NC)、 pcDNA组(转染 pcDNA)、 pcDNA-JAG1组(转染 pcDNA-JAG1)、 anti-miR-218-5p+si-NC组(共转染 anti-miR-218-5p和 si-NC)、 anti-miR-218-5p+si-JAG1组(共转染 anti-miR-218-5p和 si-JAG1)用脂质体法转染至 hPDLSCs细胞；运用实时荧光定量逆转录聚合酶链反应(qRT-PCR)法检测细胞中 miR-218-5p、Ⅰ型胶原蛋白，(Col-1)、骨钙素、 Runt相关转录因子 2(Runx2)的表达；蛋白质印迹法(Westrn blotting)检测细胞中 Jagged 1(JAG1)的蛋白表达； MTT法检测细胞活性；茜素红染色实验检测细胞的矿化结节；双荧光素酶报告基因检测实验检测细胞的荧光活性。结果与 anti-miR-NC组相比， anti-miR-218-5p组 hPDLSCs细胞培养 48、72 h时，细胞活性升高［48 h:(0.44±0.04)比( 0.62±0.06)； 72 h:(0.53±0.05)比( 0.83±0.08)； P<0.001］细胞的矿化结节明显升高， Col-1［(0.26±0.03)比( 0.74±0.07)］、骨钙素［(0.21±0.02)比( 0.54±0.05)］、 Runx-2［(0.29±0.03)比(0.61，±0.06)］蛋白相对表达量均显著升高(均 P<0.001)。与 pcDNA组相比， pcDNA-JAG1组 hPDLSCs细胞培养 48、72 h时，细胞活性显著升高［48 h:(0.42± |
| 英文摘要: |
| Objective To investigate the effect of microRNA-218-5p (miR-218-5p) on the activity and osteogenic differentiation ofhuman periodontal ligament stem cells (hPDLSCs) and to explore its mechanism, and to provide research basis for the treatment of peri.odontitis.Methods Cells were divided into anti-miR-218-5p group (transfected with anti-miR-218-5p), anti-miR-NC group (transfect. ed with anti-miR-NC), pcDNA group (transfected with pcDNA), pcDNA-JAG1 group (transfected with pcDNA-JAG1), anti-miR-218-5p+si-NC group (co-transfected with anti-miR-218-5p and si-NC), anti-miR-218-5p+si-JAG1 group (co-transfected with anti-miR-218-5p and si-JAG1), transfected into hPDLSCs cells by liposome method. Real-time fluorescence quantitative reverse transcription poly. merase chain reaction (qRT-PCR) method was used to detect miR-218-5p, type I collagen (Col-1), osteocalcin (OCN), Runt-related tran. scription factor 2 (Runx2) in cells; Western blotting method was used to detect the protein expression of Jagged 1 (JAG1) in cells; tetra.methylazolylazolyl salt microenzyme reaction colorimetric method (MTT) was used to detect cell viability; alizarin red staining test wasused to detect mineralized nodules of cells; dual luciferase reporter assay was used to detect the fluorescence activity of cells.Results Compared with the anti-miR-NC group, the cell viability of hPDLSCs in the anti-miR-218-5p group was increased at 48 and 72 h [48 h: (0.44±0.04), (0.62±0.06); 72 h: (0.53±0.05), (0.83±0.08); P<0.001]; the mineralized nodules of cells were significantly increased, and the relative expressions of Col-1, OCN, and Runx-2 proteins were significantly increased [Col-1 protein: (0.26±0.03), (0.74±0.07); OCN protein : (0.21±0.02), (0.54±0.05); Runx-2 protein: (0.29±0.03), (0.61±0.06); P<0.001]. Compared with the pcDNA group, the cell via. bility of hPDLSCs in the pcDNA-JAG1 group was significantly increased when cultured for 48 and 72 h [48 h: (0.42±0.04), (0.59±0.05); 72 h: (0.55±0.05), (0.81±0.08); P<0.001], the expression of Col-1, OCN, and Runx-2 proteins all were increased significantly [Col-1 protein: (0.34±0.03), (0.71±0.07); OCN protein: (0.23±0.02), (0.48±0.04); Runx-2 protein: (0.25±0.03), (0.56±0.05); P<0.001]. miR-218-5p can target the expression of JAG1. Inhibition of JAG1 reversibly inhibited the activity and osteogenic differentiation of hP.DLSCs by miR-218-5p.Conclusions Inhibition of miR-218-5p can promote the activity and osteogenic differentiation of human peri.odontal ligament stem cells. The mechanism is related to the targeting of JAG1. |
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