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

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7d.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(5μmol/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;Trieirbine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.

Ginsenoside Rg1 regulates the proliferation and migration of human periodontal ligament cells via Akt/eNOS signaling under nicotine stress

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.MTI assay and Transwcll 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.