中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (13): 2061-2067.doi: 10.12307/2024.121

• 牙髓及牙周膜干细胞 Dental pulp and periodontal ligament stem cells • 上一篇    下一篇

人参皂苷Rg1抑制H2O2诱导人牙周膜细胞的凋亡及自噬

杨婉蓉1，2，储  艺1，2，徐  瑶1，2，李思慧1，2，郭  玲1，2，3   

  1. 1西南医科大学口腔医学院，四川省泸州市   646000；2西南医科大学附属口腔医院修复科，四川省泸州市   646000；3口颌面修复重建和再生泸州市重点实验室，四川省泸州市   646000
  • 收稿日期:2023-02-07 接受日期:2023-03-22 出版日期:2024-05-08 发布日期:2023-08-28
  • 通讯作者: 郭玲，博士，教授，西南医科大学口腔医学院，四川省泸州市 646000；西南医科大学附属口腔医院修复科，四川省泸州市 646000；口颌面修复重建和再生泸州市重点实验室，四川省泸州市 646000
  • 作者简介:杨婉蓉，女，1995年生，四川省广安市人，西南医科大学在读硕士，主要从事口腔修复的临床与基础研究。
  • 基金资助:
    西南医科大学项目(2021ZKZD008)，项目负责人：郭玲；泸州市科技局项目(2022-GYF-11)，项目负责人：郭玲

Ginsenoside Rg1 inhibits H2O2-induced apoptosis and autophagy in human periodontal ligament cells

Yang Wanrong1, 2, Chu Yi1, 2, Xu Yao1, 2, Li Sihui1, 2, Guo Ling1, 2, 3   

  1. 1School of Stomatology of Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Department of Oral Prosthodontics, Affiliated Stomatological Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; 3Oral & Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory, Luzhou 646000, Sichuan Province, China
  • Received:2023-02-07 Accepted:2023-03-22 Online:2024-05-08 Published:2023-08-28
  • Contact: Guo Ling, MD, Professor, School of Stomatology of Southwest Medical University, Luzhou 646000, Sichuan Province, China; Department of Oral Prosthodontics, Affiliated Stomatological Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; Oral & Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory, Luzhou 646000, Sichuan Province, China
  • About author:Yang Wanrong, Master candidate, School of Stomatology of Southwest Medical University, Luzhou 646000, Sichuan Province, China; Department of Oral Prosthodontics, Affiliated Stomatological Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:
    a grant from Southwest Medical University, No. 2021ZKZD008 (to GL); a grant from Luzhou Science and Technology Bureau, No. 2022-GYF-11 (to GL)

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摘要：


文题释义：

氧化应激：当细胞内活性氧大量增多时，氧化还原平衡稳态被打破，细胞产生氧化应激，氧化应激能通过促使脂质发生过氧化，破坏蛋白质的结构、功能和损害DNA等导致细胞功能异常。
自噬：维持细胞内稳态的重要分解代谢过程，将胞浆和/或其特定内容物输送至溶酶体进行降解，由此产生的大分子成分被细胞回收利用。过量的自噬会导致细胞死亡，被称为“Ⅱ型”或“自噬”细胞死亡。


背景：课题组前期研究发现H2O2和人参皂苷Rg1能引起人牙周膜细胞活性氧水平发生变化，但活性氧和细胞凋亡、自噬的关系尚不清楚。

目的：探索人参皂苷Rg1对H2O2诱导人牙周膜细胞凋亡和自噬的作用及机制。
方法：将人牙周膜细胞分为对照组、H2O2组和H2O2+Rg1组，其中人参皂苷Rg1(50 μmol/L)预孵育24 h，H2O2处理2 h(500 μmol/L)。CCK-8法检测细胞的增殖能力；荧光探针DCFH-DA检测细胞的活性氧水平；qRT-PCR、Western blot法检测血红素加氧酶1、凋亡相关因子Caspase-3、Bax、抗凋亡因子Bcl-2、自噬相关因子Beclin-1、P62、LC3、通路相关因子PI3K、AKT、mTOR mRNA及蛋白表达。

结果与结论：①与对照组比较，H2O2组人牙周膜细胞的增殖活力下降；与H2O2组比较，H2O2+Rg1组人牙周膜细胞的增殖活力升高；②与对照组比较，H2O2组人牙周膜细胞内活性氧和血红素加氧酶1表达增加；与H2O2组比较，H2O2+Rg1组人牙周膜细胞内活性氧和血红素加氧酶1表达降低；③与对照组比较，H2O2组人牙周膜细胞内Caspase-3、Bax、Beclin-1、LC3表达升高，Bcl-2、P62表达降低；与H2O2组比较，H2O2+Rg1组人牙周膜细胞内Caspase-3、Bax、Beclin-1、LC3表达降低，Bcl-2、P62表达升高；④与对照组比较，H2O2组人牙周膜细胞内PI3K、AKT、mTOR表达降低；与H2O2组比较，H2O2+Rg1组人牙周膜细胞内PI3K、AKT、mTOR表达升高；⑤结果提示：人参皂苷Rg1能够通过清除细胞内过量活性氧，上调PI3K/AKT/mTOR通路相关因子的表达，从而抑制H2O2诱导的人牙周膜细胞凋亡和自噬。

https://orcid.org/0000-0001-5586-5245 (郭玲) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 人参皂苷Rg1, PI3K, AKT, mTOR, 活性氧, 凋亡, 自噬

Abstract: BACKGROUND: Our previous studies have confirmed that H2O2 and ginsenoside Rg1 can cause changes in reactive oxygen species levels in human periodontal ligament cells, but the correlation of reactive oxygen species with apoptosis and autophagy remains unclear.
OBJECTIVE: To explore the effect and mechanism of ginsenoside Rg1 on H2O2-induced apoptosis and autophagy of human periodontal ligament cells. 
METHODS: Human periodontal ligament cells were divided into control group, H2O2 group and H2O2+Rg1 group. Ginsenoside Rg1 (50 μmol/L) was pre-incubated for 24 hours and H2O2 was treated for 2 hours (500 μmol/L). CCK-8 assay was used to detect the proliferation ability of the cells. A fluorescent probe DCFH-DA was used to detect the reactive oxygen species level of the cells. qRT-PCR and western blot assay were used to detect heme oxygenase 1, apoptosis-related factor Caspase-3, Bax, anti-apoptotic factor Bcl-2, autophagy Beclin-1, P62, LC3, pathway-related factors phosphatidylinositol-3-kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR) mRNA and protein expression.
RESULTS AND CONCLUSION: (1) Compared with the control group, the proliferation activity of human periodontal ligament cells in the H2O2 group decreased. Compared with the H2O2 group, the proliferation activity of human periodontal ligament cells increased in the H2O2+Rg1 group. (2) Compared with the control group, the expression of reactive oxygen species and heme oxygenase-1 increased in the H2O2 group. Compared with the H2O2 group, the expression of reactive oxygen species and heme oxygenase-1 decreased in the H2O2+Rg1 group. (3) Compared with the control group, the expression of Caspase-3, Bax, Beclin-1 and LC3 increased, while the expression of Bcl-2 and P62 decreased in human periodontal ligament cells of the H2O2 group. Compared with the H2O2 group, the expressions of Caspase-3, Bax, Beclin-1 and LC3 decreased, and the expressions of Bcl-2 and P62 increased in the H2O2+Rg1 group. (4) Compared with the control group, the expressions of PI3K, AKT and mTOR decreased in human periodontal ligament cells of the H2O2 group. Compared with the H2O2 group, the expressions of PI3K, AKT and mTOR increased in human periodontal ligament cells of the H2O2+Rg1 group. (5) These results suggest that ginsenoside Rg1 can inhibit H2O2-induced apoptosis and autophagy in human periodontal ligament cells by reducing the content of reactive oxygen species and down-regulating the related factor expression of the PI3K/AKT/mTOR pathway.

Key words: ginsenoside Rg1, PI3K, AKT, mTOR, reactive oxygen species, apoptosis, autophagy

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