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肠道菌群代谢物脱氧胆酸对人脐带间充质干细胞hUC-MSCs增殖及细胞周期的影响

梁彩红 孟明耀 李欣欣 熊晶晶 李檬 刘梅 侯宗柳 黄永坤

梁彩红, 孟明耀, 李欣欣, 熊晶晶, 李檬, 刘梅, 侯宗柳, 黄永坤. 肠道菌群代谢物脱氧胆酸对人脐带间充质干细胞hUC-MSCs增殖及细胞周期的影响[J]. 昆明医科大学学报, 2023, 44(4): 23-30. doi: 10.12259/j.issn.2095-610X.S20230402
引用本文: 梁彩红, 孟明耀, 李欣欣, 熊晶晶, 李檬, 刘梅, 侯宗柳, 黄永坤. 肠道菌群代谢物脱氧胆酸对人脐带间充质干细胞hUC-MSCs增殖及细胞周期的影响[J]. 昆明医科大学学报, 2023, 44(4): 23-30. doi: 10.12259/j.issn.2095-610X.S20230402
Caihong LIANG, Mingyao MENG, Xinxin LI, Jingjing XIONG, Meng LI, Mei LIU, Zongliu HOU, Yongkun HUANG. Effect of Intestinal Flora Metabolites Deoxycholic Acid on the Proliferation and Cell Cycle of Human Umbilical Cord Mesenchymal Stem Cells[J]. Journal of Kunming Medical University, 2023, 44(4): 23-30. doi: 10.12259/j.issn.2095-610X.S20230402
Citation: Caihong LIANG, Mingyao MENG, Xinxin LI, Jingjing XIONG, Meng LI, Mei LIU, Zongliu HOU, Yongkun HUANG. Effect of Intestinal Flora Metabolites Deoxycholic Acid on the Proliferation and Cell Cycle of Human Umbilical Cord Mesenchymal Stem Cells[J]. Journal of Kunming Medical University, 2023, 44(4): 23-30. doi: 10.12259/j.issn.2095-610X.S20230402

肠道菌群代谢物脱氧胆酸对人脐带间充质干细胞hUC-MSCs增殖及细胞周期的影响

doi: 10.12259/j.issn.2095-610X.S20230402
基金项目: 云南省检验医学重点实验室开放课题基金资助项目(2017DG005)
详细信息
    作者简介:

    梁彩红(1996~),女,广东湛江人,在读硕士研究生,主要从事胃肠肝脏及微生态研究工作

    通讯作者:

    黄永坤,E-mail:hykkmyncnwd@163.com

  • 中图分类号: Q939.121

Effect of Intestinal Flora Metabolites Deoxycholic Acid on the Proliferation and Cell Cycle of Human Umbilical Cord Mesenchymal Stem Cells

  • 摘要:     目的   探讨肠道菌群代谢物次级胆汁酸DCA不同浓度和不同作用时间在体外环境下对人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)增殖和细胞周期的影响。    方法   CCK8及流式细胞仪检测在含不同浓度DCA(0.00 μmol/L、1.56 μmol/L、3.13 μmol/L、6.25 μmol/L、12.50 μmol/L、25.00 μmol/L、50.00 μmol/L、100.00 μmol/L、200.00 μmol/L、400.00 μmol/L、800.00 μmol/L)干细胞培养基中培养hUC-MSCs 24 h、48 h、72 h后,干细胞增殖及细胞周期的变化。    结果   脱氧胆酸对hUC-MSCs增殖的影响体现在DCA剂量和作用时间两方面(两者交互作用,F = 6.622,P < 0.001);当浓度一定时,时间越长,细胞增殖越多;当作用时间一定时,浓度越高,细胞增殖抑制。浓度一定时,随时间的增加,细胞阻滞在G0/G1期的比例增加(P < 0.001);不同作用时间DCA对 hUC-MSCs的细胞周期表现为低浓度组细胞阻滞在G0/G1期和S期,高浓度组细胞阻滞在S期和G2/M期(P < 0.001)。    结论   体外环境下,DCA对hUC-MSCs增殖及细胞周期的影响与浓度和作用时间相关。
  • 图  1  脱氧胆酸不同浓度及不同作用时间对 hUC-MSCs 增殖的影响

    A:脱氧胆酸不同浓度及不同作用时间对 hUC-MSCs 增殖的影响;B:不同浓度脱氧胆酸作用 24 h 时 hUC-MSCs增殖情况;C:不同浓度脱氧胆酸作用48 h时hUC-MSCs增殖情况;D:不同浓度脱氧胆酸作用72 h时hUC-MSCs 增殖情况。各浓度组与0.00 μmol/L浓度组比较,ns表示差异无统计学意义,*表示 P < 0.05,**表示 P < 0.01,***表示 P < 0.001,****表示 P < 0.0001。

    Figure  1.  Effects of DCA with different concentration and different time on the proliferation of hUC- MSCs

    图  2  脱氧胆酸不同浓度及不同作用时间 hUC-MSCs 细胞周期分布情况

    Figure  2.  Cell cycle proportion of hUC-MSCs with different concentration and different time of DCA

    图  3  不同浓度及不同作用时间脱氧胆酸刺激后流式细胞技术检测 hUC-MSCs 细胞周期代表图例

    A~D:24 h时脱氧胆酸0.00 μmol/L~400.00 μmol/L hUC-MSCs细胞周期分布情况;E~H:48 h时脱氧胆酸0.00 μmol/L~400.00 μmol/L hUC-MSCs细胞周期分布情况;I~L:72 h时脱氧胆酸0.00 μmol/L~400.00 μmol/L hUC-MSCs细胞周期分布情况。

    Figure  3.  Cell cycle of hUC-MSCs tested by flow cytometry after different concentration and different time of DCA

    表  1  脱氧胆酸不同浓度及不同作用时间hUC-MSCs吸光度值(OD450nm值)($\bar x \pm s $

    Table  1.   Absorbance (OD450nm) of hUC-MSCs at different dose and time of DCA ($\bar x \pm s $

    脱氧胆酸浓度(μmol/L)样本数24 h48 h72 h
    0.00(对照组) 45 0.61 ± 0.12 0.77 ± 0.14* 0.97 ± 0.32○*
    1.56 45 0.51 ± 0.07 0.79 ± 0.19* 1.23 ± 0.24○*
    3.13 45 0.54 ± 0.08 0.79 ± 0.21* 1.01 ± 0.31○*
    6.25 45 0.56 ± 0.09 0.80 ± 0.16* 1.05 ± 0.33○*
    12.50 45 0.55 ± 0.05 0.81 ± 0.07* 1.18 ± 0.28○*
    25.00 45 0.55 ± 0.07 0.78 ± 0.13* 1.21 ± 0.20○*
    50.00 45 0.57 ± 0.08 0.77 ± 0.03* 1.07 ± 0.24○*
    100.00 45 0.58 ± 0.08 0.75 ± 0.16* 1.05 ± 0.23○*
    200.00 44 0.58 ± 0.09 0.72 ± 0.16 0.84 ± 0.26*
    400.00 45 0.50 ± 0.10 0.54 ± 0.19 0.53 ± 0.17
    800.00 45 0.49 ± 0.12 0.48 ± 0.18 0.49 ± 0.18
      与24 h比较,*P < 0.05;与48 h比较,P < 0.05。
    下载: 导出CSV

    表  2  脱氧胆酸不同浓度及不同作用时间 hUC-MSCs 细胞周期比例(%)

    Table  2.   Cell cycle ratio of hUC-MSCs with different concentration and different time of DCA (%)

    时间
    (h)
    分期0.00
    μmol/L
    1.56
    μmol/L
    25.00
    μmol/L
    400.00
    μmol/L
    24 G0/G1期 77.60 82.33* 83.77* 48.17*
    S期 16.48 17.67 16.23 51.83*
    G2/M期 5.91 0.00* 0.00* 0.00*
    48 G0/G1期 87.07 92.85*△ 85.73*△ 42.99*△
    S期 5.45 2.92*△ 14.27*△ 50.89*
    G2/M期 7.48 4.23*△ 0.00* 6.12*△
    72 G0/G1期 97.60 98.19*△ 98.41*△ 73.14*△
    S期 2.40 1.81*△ 1.59*△ 22.70*△
    G2/M期 0.00 0.00 0.00 4.16*△
      与0.00 µmol/L比较,*P < 0.0001;与24 h比较,P < 0.0001。
    下载: 导出CSV
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  • 收稿日期:  2023-01-05
  • 网络出版日期:  2023-04-19
  • 刊出日期:  2023-04-25

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