人参皂苷Rg1通过Akt/eNOS信号调控尼古丁胁迫下人牙周膜细胞的增殖和迁移

doi:10.19439/j.sjos.2017.01.009

摘要/Abstract

摘要： 目的探讨人参皂苷Rg1对尼古丁胁迫下的人牙周膜细胞（human periodontal ligament cells,HPDLCs）增殖和迁移的影响及分子机制。方法采用组织块法分离培养HPDLCs,尼古丁（500 ng/mL）胁迫培养细胞7 d。第3天分别进行人参皂苷Rg1（0.01 μmol/L）处理、人参皂苷Rg1与PI3K抑制剂LY294002（0.5 μmol/L）共处理、人参皂苷Rg1与Akt抑制剂Tricirbine（5 μmol/L）共处理、人参皂苷Rg1与eNOS抑制剂 L-NAME（1 mmol/L）共处理（Rg1+L-NAME组）,持续处理至第7天。采用MTT法和Transwell法检测各处理组HPDLCs活力变化和细胞迁移率,并使用实时定量PCR与Western印迹法检测PI3K/Akt/eNOS信号蛋白变化,采用SPSS20.0软件包对数据进行统计学分析。结果尼古丁抑制HPDLCs的增殖和迁移并显著上调PI3K表达,抑制Akt和eNOS表达;人参皂苷Rg1减缓尼古丁对细胞增殖和迁移抑制作用以及尼古丁对Akt和eNOS表达的抑制作用;Tricirbine与L-NAME衰减人参皂苷Rg1对尼古丁的抑制作用。结论人参皂苷Rg1通过Akt/eNOS信号调控尼古丁胁迫下HPDLCs的增殖和迁移。

关键词: 牙周膜细胞, 尼古丁, 人参皂苷Rg1, Akt/eNOS信号

Abstract: PURPOSE: To explore the effects and molecular mechanisms of ginsenoside Rg1 on the proliferation and migration of human periodontal ligament cells (HPDLCs) under nicotine stress. METHODS: HPDLCs were isolated and cultured by method of explant cell culture. The cells were cultured under nicotine stress for 7 days, and treated respectively with ginsenoside Rg1 (0.01 μmol/L), ginsenoside Rg1 and LY294002 (PI3K inhibitor, 0.5 μmol/L), ginsenoside Rg1 and Tricirbine (Akt inhibitor, 5 μmol/L), ginsenoside Rg1 and L-NAME (Akt inhibitor, 1 mmol/L) from 3rd day after nicotine stress to 7th day. MTT assay and Transwell assay were used to evaluate the proliferation and migration of HPDLCs in each group. Western blot and quantitative real-time PCR methods were used for testing the changes of PI3K/Akt/eNOS signaling expression. SPSS 20.0 software package was used for statistical analysis. RESULTS: The proliferation and migration were significantly inhibited by nicotine treatment. PI3K levels were upregulated, but Akt1/2 and eNOS levels were remarkedly reduced by nicotine. Ginsenoside Rg1 attenuated the effects of nicotine on proliferation, migration and Akt/eNOS signaling. Tricirbine and L-NAME could reduce the inhibitory effects of ginsenoside Rg1 toward nicotine. CONCLUSIONS: Ginsenoside Rg1 regulates the proliferation and migration of HPDLCs under nicotine stress via Akt/eNOS signaling.

Key words: Periodontal ligament cells, Nicotine, Ginsenoside Rg1, Akt/eNOS signaling

中图分类号:

R781.4

刘彩宏, 杜莉. 人参皂苷Rg1通过Akt/eNOS信号调控尼古丁胁迫下人牙周膜细胞的增殖和迁移[J]. 上海口腔医学, 2017, 26(1): 42-47.

LIU Cai-hong, DU Li. Ginsenoside Rg1 regulates the proliferation and migration of human periodontal ligament cells via Akt/eNOS signaling under nicotine stress[J]. Shanghai Journal of Stomatology, 2017, 26(1): 42-47.

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