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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/m L)胁迫培养细胞7 d。第3天分别进行人参皂苷Rg1(0.01μmol/L)处理、人参皂苷Rg1与PI3K抑制剂LY294002(0.5μmol/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e NOS抑制剂L-NAME(1 mmol/L)共处理(Rg1+LNAME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/e NOS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e NOS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e NOS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/e NOS信号调控尼古丁胁迫下HPDLCs的增殖和迁移。
Objective: To investigate the effect of ginsenoside Rg1 on proliferation and migration of human periodontal ligament cells (HPDLCs) under nicotine stress and its molecular mechanism. Methods: HPDLCs were isolated and cultured by tissue block method. Nicotine (500 ng / m L) stress cells were cultured for 7 days. Ginsenoside Rg1 and PI3K inhibitor LY294002 (0.5μmol / L) were co-treated with Ginsenoside Rg1 (0.01μmol / L) on day 3, and co-treated with Akt inhibitor Tricirbine (5μmol / L) Saponin Rg1 and e NOS inhibitor L-NAME (1 mmol / L) co-treated (Rg1 + LNAME group) and continued to the seventh day. The cell viability and the viability of HPDLCs in each treatment group were detected by MTT assay and Transwell assay. The changes of PI3K / Akt / e NOS signal proteins were detected by real-time PCR and Western blotting. The data were analyzed by SPSS 20.0 software package . RESULTS: Nicotine could inhibit the proliferation and migration of HPDLCs and up-regulate the expression of PI3K and Akt and eNOS. Nicotinamide ginsenoside Rg1 could inhibit the inhibitory effect of nicotine on the cell proliferation and migration as well as the inhibitory effect of nicotine on Akt and e NOS expression. Tricirbine and L- NAME attenuates the inhibitory effect of ginsenoside Rg1 on nicotine. Conclusion: Ginsenoside Rg1 regulates the proliferation and migration of HPDLCs under Akt / e NOS signaling.