三七总皂苷对人根尖牙乳头干细胞增殖的影响

钱石兵; 孟明耀; 于鸿滨; 段开文; 李昌全; 夏志刚

doi:10.12259/j.issn.2095-610X.S20221018

三七总皂苷对人根尖牙乳头干细胞增殖的影响

doi: 10.12259/j.issn.2095-610X.S20221018

1.
昆明医科大学附属延安医院口腔科，云南昆明　650051
2.
中心实验室，云南昆明　650051

基金项目: 云南省生物医药重大科技专项（202102AA100007）；云南省高层次卫生计生技术人才培养计划基金资助项目（H-2018089）；昆明市卫生健康委员会卫生科研课题项目（2021-08-02-001）

详细信息

作者简介:
钱石兵（1996～），男，云南昆明人，在读硕士研究生，研究方向为牙周和口腔黏膜疾病的预防

通讯作者:
夏志刚，E-mail：xiayang928@sina.com

中图分类号: R285.5
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出版历程
- 收稿日期: 2022-06-11
- 网络出版日期: 2022-10-09
- 刊出日期: 2022-10-31

Effect of Panax Noto-ginseng Saponins on the Proliferation of Human Stem Cells from Apical Papilla

1.
Dept. of Stomatology，Yan’an Hospital，Kunming City，The Affiliated Yan’an Hospital，Kunming Medical University，Kunming Yunnan 650051
2.
Central Laboratory，Yan’an Hospital, Kunming City，The Affiliated Yan’an Hospital，Kunming Medical University， Kunming Yunnan 650051，China

摘要

摘要: 目的探究三七总皂苷（panax noto-ginseng saponins，PNS）对人根尖牙乳头干细胞（human stem cells from apical papilla, hSCAPs）增殖的影响。方法收集牙乳头组织、贴壁法分离培养原代细胞，流式细胞术鉴定hSCAPs表面标志物，诱导成骨及成脂鉴定hSCAPs多向分化潜能；配制25 mg/L、50 mg/L、100 mg/L、200 mg/L的PNS培养基，CCK-8法检测不同浓度PNS下hSCAPs的增殖速率，流式细胞术检测最佳促进增殖组与对照组的细胞周期差异。结果原代分离培养成功的hSCAPs符合间充质干细胞来源，相关表面标记物高表达，可向成脂和成骨分化；与对照和其他实验组比较，25 mg/LPNS促进hSCAPs增殖效果最佳,差异具有统计学意义（P < 0.05）。结论 25 mg/LPNS促进hSCAPs增殖能力的效果最佳，提示hSCAPs对PNS具有很强的敏感性。
- 三七总皂苷 /
- 人根尖牙乳头干细胞 /
- 细胞增殖 /
- 细胞周期
Abstract: Objective To explore the effects of which Panax noto-ginseng saponins （PNS） influence the proliferation of human stem cells from apical papilla （hSCAPs）. Methods We collected apical papilla and separated primary cells with wall methods. Flow cytometry was used to identify the surface markers of hSCAPs, the multi-directional-differentiation potential of hSCAPs was identified by inducing bone and fat differentiation. PNS-culture-medium with 25 mg/L, 50 mg/L, 100 mg/L, and 200 mg /L was prepared, CCK-8 method was used to detect the proliferation rate of hSCAPs under different concentrations of PNS, and the changes of the cell cycle at the best proliferation concentration were observed. Results hSCAPs was cultivated successfully by primary separation and belong to the source of mesenchymal stem cells. The related surface markers were highly expressed and induced to fat differentiation and bone differentiation. Compared with the control and other experimental groups, PNS with 25 mg/L promoted highly hSCAPs proliferation. The difference was statistically significant （P < 0.05）. Conclusions PNS with 25 mg/L has the best effect for boosting hSCAPs proliferation abilities. It is prompted that hSCAPs has a strong sensitivity to PNS, that is, a low concentration can significantly promote the proliferation of hSCAPs.
- Panax notoginseng saponins /
- Human apical papilla stem cells /
- Cell proliferation /
- Cell cycle

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图 1 hSCAPs的培养及多向分化能力鉴定

A：hSCAPs原代细胞；B：细胞传代培养；C：hSCAPs成骨诱导后茜素红染色；D：hSCAPs成骨对照组茜素红染色；E：hSCAPs成脂诱导后油红O染色；F：hSCAPs成脂对照组油红O染色。

Figure 1. Culture of hSCAPs and identification of multi -directional differentiation ability

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图 2 hSCAPs流式细胞鉴定结果

　　注：CD73 、CD90 和CD105均为间充质干细胞表面标志物，流式结果显示均阳性高表达，CD34、CD45为造血细胞标记物，CD19、CD11b、HLA-DR是免疫细胞表面标记物均呈呈阴性。

Figure 2. Identification of hSCAPs surface antigen by flow cytometry

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图 3 不同浓度PNS下hSCAPs的生长曲线

Figure 3. The growth curve of hSCAPs under different concentrations of PNS

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图 4 对照组细胞周期分布

Figure 4. Cell cycle distribution of control group

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图 5 25 mg/L组细胞周期分布

Figure 5. Cell cycle distribution of 25 mg/L group

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图 6 细胞增殖指数差异

^*P < 0.05。

Figure 6. Difference in cell proliferation index

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表 1 不同浓度PNS下hSCAPs增殖的差异

Table 1. Differences in HSCAPS proliferation under different concentrations of PNS OD 值，$ \stackrel{-}{X} $ ± S

组别	第1天	第3天	第5天	第7天	第9天
对照组	0.199 ± 0.005	0.275 ± 0.005	0.731 ± 0.006	0.984 ± 0.023	1.975 ± 0.062
25 mg/L	0.126 ± 0.006^*	0.442 ± 0.01^*	1.385 ± 0.067^*	1.549 ± 0.017^*	2.784 ± 0.057^*
50 mg/L	0.075 ± 0.005^*^#	0.407 ± 0.011^*^#	1.058 ± 0.007^*^#	1.146 ± 0.016^*^#	2.443 ± 0.015^*^#
100 mg/L	0.044 ± 0.003^*^#^&	0.245 ± 0.009^*^#^&	0.213 ± 0.009^*^#^&	0.187 ± 0.003^*^#^&	0.158 ± 0.007^*^#^&
200 mg/L	0.024 ± 0.002^*^#^&^△	0.192 ± 0.003^*^#^&^△	0.168 ± 0.005^*^#^&^△	0.126 ± 0.003^*^#^&^△	0.079 ± 0.013^*^#^&^△
F	490.006	358.621	598.128	3 552.492	2 174.635
P	0.000	0.000	0.000	0.000	0.000
与对照组比较，^*P < 0.05；实验组两两比较，与25 mg/L组比较，^#P < 0.05；与50 mg/L组比较，^&P < 0.05；与100 mg/L组比较，^△P < 0.05。

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