丹参水溶性成分拮抗脂多糖诱导HTR8/SVneo滋养层细胞凋亡的作用机制

Mechanism of Antagonistic Action of Soluble Components of Salvia Miltiorrhiza on Apoptosis of HTR8/SVneo Induced by Lipopolysaccharide

关键词:  脂多糖  HTR8/SVneo  丹参酚酸B  丹参酚酸C  紫草酸  细胞凋亡

Key words:  lipopolysaccharide  HTR8/SVneo  salvianolic acid B  salvianolic acid C  lithospermic acid  apoptosis

摘要:

目的 探讨丹参水溶性成分拮抗脂多糖(lipopolysaccharide，LPS)诱导HTR8/SVneo细胞凋亡的相关作用机制。方法 以不同浓度(0，100，200，400，800，1 600 ng·mL^-1)LPS处理HTR8/SVneo细胞，采用实时定量聚合酶链式反应验证HTR 8/SVneo细胞炎症模型的建立；细胞活力检测试剂盒检测各组细胞活力；划痕试验评估各组细胞向损伤区域迁移能力；流式细胞术检测各组细胞凋亡率并分析各组细胞线粒体膜电位。结果 200 ng·mL^-1 LPS干预后HTR8/SVneo细胞的NLRP3炎症小体指标及炎症因子表达量增高(P<0.05)。与LPS组相比，丹酚酸B(0.08 μmol·L^-1)、丹酚酸C(0.4 μmol·L^-1)、紫草酸(0.08 μmol·L^-1)干预后HTR8/SVneo细胞活性明显升高(P<0.01)。与LPS组比较，丹参水溶物质组凋亡率显著降低(P<0.05或P<0.01)，线粒体膜电位水平显著升高。结论 丹参水溶物质能够提高LPS干预后HTR8/SVneo细胞活力，改善细胞迁移能力，提高细胞线粒体膜电位，进而抑制细胞凋亡。

Abstract:

OBJECTIVE To explore the anti-apoptosis mechanism of Salvia miltiorrhiza soluble substances on HTR8/SVneo induced by lipopolysaccharide(LPS). METHODS HTR8/SVneo cells were treated with LPS of different concentrations(0, 100, 200, 400, 800, 1 600 ng·mL^-1), and Real-time quantitative PCR was used to verify the establishment of HTR8/SVneo cell inflammation model. Cell viability in each group was detected by cell counting Kit-8. Cells migration ability to the injured area was evaluated by scratch test. Flow cytometry was used to detect cell apoptosis rate in each group and analyze mitochondrial membrane potential.RESULTS After 200 ng·mL^-1 LPS intervention, the transcriptional level of NLRP3 inflammasome and inflammatory cytokines in HTR8/SVneo cells were increased(P<0.05). Compared with LPS group, the activity of HTR8/SVneo was significantly increased after the intervention of salvianolic acid B(0.08 μmol·L^-1), salvianolic acid C(0.4 μmol·L^-1) and lithospermic acid(0.08 μmol·L^-1)(P<0.01). Compared with LPS group, the apoptosis rate of Salvia miltiorrhiza soluble substance intervention group was significantly decreased(P<0.05 or P<0.01), the level of mitochondrial membrane potential was significantly increased. CONCLUSION Salvia miltiorrhiza soluble substance can enhance the cell viability of HTR8/SVneo induced by LPS, improve cell migration ability, enhance mitochondrial membrane potential, and then inhibit cell apoptosis.