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中华细胞与干细胞杂志(电子版) ›› 2024, Vol. 14 ›› Issue (04) : 229 -235. doi: 10.3877/cma.j.issn.2095-1221.2024.04.005

论著

银杏内酯B通过调控miR-24-3p对人牙周膜干细胞增殖、成骨分化的影响
李彦浇1, 梁雷1, 金钫1, 王智伟1,()   
  1. 1. 710032 西安,口颌系统重建与再生全国重点实验室 国家口腔疾病临床医学研究中心 陕西省口腔疾病临床医学研究中心 第四军医大学口腔医院正畸科
  • 收稿日期:2024-02-26 出版日期:2024-08-01
  • 通信作者: 王智伟
  • 基金资助:
    国家自然科学基金(82170988)

Effects of ginkgolide B on proliferation and osteogenic differentiation of human perimembrane stem cells by regulating miR-24-3p

Yanjiao Li1, Lei Liang1, Fang Jin1, Zhiwei Wang1,()   

  1. 1. Department of Orthodontics, School of Stomatology, the Fourth Military Medical University, Shaanxi Clinical Research Center for Oral Diseases, National Clinical Research Center for Oral Diseases, State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Xi'an 710032, China
  • Received:2024-02-26 Published:2024-08-01
  • Corresponding author: Zhiwei Wang
引用本文:

李彦浇, 梁雷, 金钫, 王智伟. 银杏内酯B通过调控miR-24-3p对人牙周膜干细胞增殖、成骨分化的影响[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 229-235.

Yanjiao Li, Lei Liang, Fang Jin, Zhiwei Wang. Effects of ginkgolide B on proliferation and osteogenic differentiation of human perimembrane stem cells by regulating miR-24-3p[J/OL]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2024, 14(04): 229-235.

目的

探究银杏内酯B (GB)调控miR-24-3p对人牙周膜干细胞增殖和成骨分化的影响。

方法

体外分离培养人牙周膜干细胞,使用10、25、50 μmol/L GB处理细胞;按照脂质体法将anti-miR-NC、anti-miR-24-3p转染细胞内;将miR-NC、miR-24-3p转染细胞内,用50 μmol/L GB处理。用MTT检测细胞增殖。将各组细胞进行成骨诱导后,试剂盒检测ALP活性;Western blot检测PCNA、Osx、OPN和OCN蛋白表达;RT-qPCR检测miR-24-3p表达水平。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验。

结果

GB呈剂量依赖性增加人牙周膜干细胞增殖活性[48 h:(0.52 ± 0.05、0.64 ± 0.05、0.80 ± 0.08比0.33 ± 0.05);72 h:(0.72 ± 0.05、0.85 ± 0.04、1.08 ± 0.11比0.51 ± 0.06)]、ALP活性(0.45 ± 0.06、0.75 ± 0.08、1.12 ± 0.08比0.27 ± 0.05)、PCNA (0.41 ± 0.05、0.57 ± 0.06、0.78 ± 0.06比0.26 ± 0.04)、Osx (0.45 ± 0.06、0.68 ± 0.05、0.79 ± 0.03比0.33 ± 0.04)、OPN(0.56 ± 0.06、0.75 ± 0.05、0.92 ± 0.06比0.42 ± 0.05)、OCN蛋白表达(0.48 ± 0.05、0.59 ± 0.04、0.76 ± 0.08比0.30 ± 0.06),降低miR-24-3p表达水平(0.35 ± 0.04、0.52 ± 0.06、0.87 ± 0.06比1.00 ± 0.09)(P均< 0.05)。干扰miR-24-3p可增加48、72 h时人牙周膜干细胞增殖活性[(0.57 ± 0.06比0.35 ± 0.04)、(0.90 ± 0.10比0.47 ± 0.06)]、ALP活性(0.68 ± 0.05比0.30 ± 0.04)、PCNA (0.47 ± 0.05比0.29 ± 0.04)、Osx (0.50 ± 0.04比0.35 ± 0.04)、OPN (0.62 ± 0.05比0.39 ± 0.03)、OCN蛋白表达(0.75 ± 0.07比0.42 ± 0.06) (P均< 0.05)。与GB+miR-NC比较,GB+miR-24-3p干预的细胞48、72 h的OD值[(0.67 ± 0.05比0.82 ± 0.07)、(0.84 ± 0.05比1.10 ± 0.09)]、ALP活性(0.56 ± 0.04比1.15 ± 0.15)、PCNA (0.46 ± 0.06比0.79 ± 0.07)、Osx(0.52 ± 0.03比0.80 ± 0.05)、OPN (0.63 ± 0.04比0.91 ± 0.09)、OCN蛋白表达(0.45 ± 0.06比0.78 ± 0.08)降低(P < 0.05)。

结论

GB可能通过降低miR-24-3p促进人牙周膜干细胞增殖和成骨分化。

Objective

To investigate the effect of ginkgolide B (GB) regulating miR-24-3p on human peripheral stem cells' proliferation and osteogenic differentiation.

Methods

Human periodontal stem cells were isolated and cultured in vitro. The cells were treated with 10, 25, 50 μmol/L GB. Anti-miR-NC and anti-miR-24-3p were transfected into cells by liposome method, and miR-NC and miR-24-3p were transfected into cells and treated with 50 μmol/L GB. MTT detected cell proliferation; After osteogenic induction of cells with different treatments, the ALP activity was detected by the kit; the protein expressions of PCNA, Osx, OPN and OCN were detected by Western blot; and RT-qPCR detected the expression level of miR-24-3p.

Results

GB increased the proliferative activity (48 h, 72 h) [ (0.52 ± 0.05, 0.64 ± 0.05, 0.80 ± 0.08 vs 0.33 ± 0.05), (0.72 ± 0.05, 0.85 ± 0.04, 1.08 ± 0.11 vs 0.51 ± 0.06) ], ALP activity, PCNA (0.45 ± 0.06, 0.75 ± 0.08, 1.12 ± 0.08 vs 0.27 ± 0.05), PCNA (0.41 ± 0.05, 0.57 ± 0.06, 0.78 ± 0.06 vs 0.26 ± 0.04), Osx (0.45 ± 0.06, 0.68 ± 0.05, 0.79 ± 0.03 vs 0.33 ± 0.04 ), OPN (0.56 ± 0.06, 0.75 ± 0.05, 0.92 ± 0.06 vs 0.42 ± 0.05), OCN (0.48 ± 0.05, 0.59 ± 0.04, 0.76 ± 0.08 vs 0.30 ± 0.06) protein expression of human periodontal ligament stem cells in a dose-dependent manner, and decreased the expression level of miR-24-3p (0.35 ± 0.04, 0.52 ± 0.06, 0.87 ± 0.06 vs 1.00 ± 0.09 ) (P < 0.05). Interfering with miR-24-3p increased the proliferative activity [48 h: (0.57 ± 0.06 vs 0.35 ± 0.04) ; 72 h (0.90 ± 0.10 vs 0.47 ± 0.06) ], ALP activity (0.68 ± 0.05 vs 0.30 ± 0.04 ), PCNA (0.47 ± 0.05 vs 0.29 ± 0.04), Osx (0.50 ± 0.04 vs 0.35 ± 0.04), OPN (0.62 ± 0.05 vs 0.39 ± 0.03 ), OCN (0.75 ± 0.07 vs 0.42 ± 0.06) protein expressions of human periodontal ligament stem cells (all P < 0.05). Compared with GB+miR-NC group, the OD value [48 h: (0.67 ± 0.05 vs 0.82 ± 0.07) ; 72 h: (0.84 ± 0.05 vs 1.10 ± 0.09) ], ALP activity (0.56 ± 0.04 vs 1.15 ± 0.15), PCNA (0.46 ± 0.06 vs 0.79 ± 0.07), Osx (0.52 ± 0.03 vs 0.80 ± 0.05), OPN (0.63 ± 0.04 vs 0.91 ± 0.09 ), OCN (0.45 ± 0.06 vs 0.78 ± 0.08) protein expression of GB+miR-24-3p group decreased (all P < 0.05) .

Conclusion

GB may promote proliferation and osteogenic differentiation of human periodontal stem cells by decreasing miR-24-3p.

表1 引物序列信息
图1 显微镜下观察人牙周膜干细胞的形态(×200)
图2 人牙周膜干细胞的流式细胞术鉴定结果
图3 Western blot检测银杏内酯B对人牙周膜干细胞增殖PCNA蛋白表达的影响
表2 不同浓度的GB对人牙周膜干细胞增殖的影响( ± s
图4 Western blot检测银杏内酯B对人牙周膜干细胞Osx、OPN和OCN蛋白表达的影响
表3 不同浓度的GB对人牙周膜干细胞成骨分化的影响( ± s
表4 不同浓度的银杏内酯B对人牙周膜干细胞内miRNA的影响( ± s
图5 Western blot检测干扰miR-24-3p对人牙周膜干细胞对PCNA、Osx、OPN和OCN蛋白表达的影响
表5 干扰miR-24-3p对人牙周膜干细胞增殖和成骨分化的影响( ± s
图6 Western blot检测银杏内酯B和miR-24-3p干预对PCNA、Osx、OPN和OCN蛋白表达的影响
表6 GB通过调控miR-24-3p对人牙周膜干细胞增殖和成骨分化的影响( ± s
1
Tomokiyo A, Wada N, Maeda H. Periodontal ligament stem cells: regenerative potency in periodontium[J]. Stem Cells Dev, 2019, 28(15):974-985.
2
Liu J, Ruan J, Weir MD, et al. Periodontal bone-ligament-cementum regeneration via scaffolds and stem cells[J]. Cells, 2019, 8(6):537. doi: 10.3390/cells8060537.
3
Calabrese EJ. Human periodontal ligament stem cells and hormesis: enhancing cell renewal and cell differentiation[J]. Pharmacol Res, 2021, 173:105914. doi: 10.1016/j.phrs.2021.105914
4
Li H, Zhang W, Wu Z, et al. Theoretical design, preparation, and evaluation of Ginkgolide B molecularly imprinted polymers[J]. J Sep Sci, 2020, 43(2):514-523.
5
谭雪锋, 孙冰晶, 朱倩, 等. 银杏内酯B体外诱导神经干细胞向多巴胺能神经元分化[J].解剖学杂志, 2016, 39(1):10-13.
6
张爱平, 高田, 胡玉亮. 银杏内酯B调控PAX6基因表达对人骨髓间充质干细胞增殖和成骨分化能力的影响[J]. 中国药师, 2021, 24(9):1612-1616.
7
Naqvi AR, Slots J. Human and herpesvirus microRNAs in periodontal disease[J]. Periodontol 2000, 2021, 87(1):325-339.
8
Cao T, Zhen XC. Dysregulation of miRNA and its potential therapeutic application in schizophrenia[J]. CNS Neurosci Ther, 2018, 24(7):586-597.
9
Li Z, Sun Y, Cao S, et al. Downregulation of miR-24-3p promotes osteogenic differentiation of human periodontal ligament stem cells by targeting SMAD family member 5[J]. J Cell Physiol, 2019, 234(5):7411-7419.
10
Trubiani O, Pizzicannella J, Caputi S, et al. Periodontal ligament stem cells: current knowledge and future perspectives[J]. Stem Cells Dev, 2019, 28(15):995-1003.
11
Xu XY, Li X, Wang J, et al. Concise review: periodontal tissue regeneration using stem cells: strategies and translational considerations[J]. Stem Cells Transl Med, 2019, 8(4):392-403.
12
Han Y. High concentrations of calcium suppress osteogenic differentiation of human periodontal ligament stem cells in vitro[J]. J Dent Sci, 2021, 16(3):817-824.
13
李艳萍, 张立虎, 吴红雁, 等. 银杏叶功效群组分及其功效的研究进展[J]. 食品研究与开发, 2020, 41(15):182-187.
14
Huang J, Yang J, Zou X, et al. Ginkgolide B promotes oligodendrocyte precursor cell differentiation and survival via Akt/CREB/bcl-2 signaling pathway after white matter lesion[J]. Exp Biol Med (Maywood), 2021, 246(10):1198-1209.
15
Zheng PD, Mungur R, Zhou HJ, et al. Ginkgolide B promotes the proliferation and differentiation of neural stem cells following cerebral ischemia/reperfusion injury, both in vivo and in vitro[J]. Neural Regen Res, 2018, 13(7):1204-1211.
16
Zhu B, Xue F, Zhang C, et al. Ginkgolide B promotes osteoblast differentiation via activation of canonical Wnt signalling and alleviates osteoporosis through a bone anabolic way[J]. J Cell Mol Med, 2019, 23(8):5782-5793.
17
Wang YL, Lee CC, Shen YC, et al. Evading immune surveillance via tyrosine phosphorylation of nuclear PCNA[J]. Cell Rep, 2021, 36(8):109537. doi: 10.1016/j.celrep.2021.109537.
18
Ha SH, Choung PH. MSM promotes human periodontal ligament stem cells differentiation to osteoblast and bone regeneration[J]. Biochem Biophys Res Commun, 2020, 528(1):160-167.
19
Aljohani H, Senbanjo LT, Chellaiah MA. Methylsulfonylmethane increases osteogenesis and regulates the mineralization of the matrix by transglutaminase 2 in SHED cells[J]. PLoS One, 2019, 14(12):e0225598. doi: 10.1371/journal.pone.0225598.
20
魏蓉, 李新月. miRNA与TLRs信号通路在牙周病发生发展过程中作用机制的研究进展[J]. 北京口腔医学, 2020, 28(6):355-358.
21
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, 2547.
22
Yan GQ, Wang X, Yang F, et al. MicroRNA-22 promoted osteogenic differentiation of human periodontal ligament stem cells by targeting HDAC6[J]. J Cell Biochem, 2017, 118(7):1653-1658.
23
庞鸣, 韦红霞, 陈茜. 长链非编码RNA钾离子电压门控通道亚家族Q成员1重叠转录本1通过靶向miR-24-3p调控人牙周膜干细胞增殖和成骨分化[J]. 华西口腔医学杂志, 2021, 39(5):547-554.
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