参白解毒方对溃疡性结肠炎肠黏膜上皮细胞保护作用机制研究

Explore protective mechanism of Shenbai Jiedu decoction on colon epithelial cells of ulcerative colitis

目的:研究参白解毒方(SBJDF)对溃疡性结肠炎肠黏膜上皮细胞的保护作用及机制。方法:使用葡聚糖硫酸钠(DSS)刺激人正常结直肠黏膜上皮细胞(FHC),模拟溃疡性结肠炎黏膜上皮细胞的损伤,倒置显微镜观察参白解毒方对PHC损伤模型细胞形态的影响通过MTT法检测DSS对细胞活力的影响,5-乙炔基-2’-脱氧尿苷(EdU)检测参白解毒方对细胞增殖能力的影响,流式细胞仪检测FHC细胞的周期分布变化,通过蛋白免疫印迹法检测细胞周期相关蛋白Cyclin A2、CDK1的表达。结果:DSS能够抑制FHC细胞活力,下调FHC细胞的Cyclin A2、CDK1蛋白表达,将FHC细胞周期阻滞于G_(0)/G_(1)期,抑制细胞增殖;参白解毒方给药后能减少DSS细胞毒性,增加FHC细胞的Cyclin A2、CDK1蛋白表达,进而逆转DSS导致的FHC细胞G_(0)/G_(1)期阻滞,增加细胞活力。结论:参白解毒方可能通过维持细胞周期稳态发挥对溃疡性结肠炎肠黏膜上皮细胞的保护作用。

Objective:To explore the protective mechanism of Shenbai Jiedu decoction(SBJDF)on ulcerative colitis.Methods:Dextran sodium sulfate(DSS)was used to stimulate normal fetal human colon(FHC)cells to simulate the ulcerative colitis mucosal epithelial cells.After treatment with SBJDF,cell morphology was observed by inverted microscopy,The MTT assay was used to detect the effect of SBJDF on the cell viability of FHC cells,EdU was used to detect the cell proliferation ability,The cell cycle distribution changes was observed by flow cytometry,The expression levels of proteins Cyclin A2,CDK1 were determined by Western blot analysis.Results:DSS decreased viability of FHC cells,DSS induced FHC cell cycle arrest in G_(0)/G_(1) phase and down-regulated the expression of Cyclin A2 and CDK1,inhibited cell proliferation.After treatment with DSS,SBJDF can inhibit the decrease of FHC cell viability induced by DSS and reverse the down-regulation of Cyclin A2 and CDK1,thereby alleviate the G_(0)/G_(1) phase block of FHC cells caused by DSS.Conclusion:SBJDF may protect colorectal mucosa cells from inflammatory bowel disease by maintaining cell cycle homeostasis.