清肠栓对LPS诱导Caco-2细胞损伤的保护作用

Protective effects of Qingchang Suppository on LPS-induced Caco-2 cell injury

目的探讨清肠栓(QCS)含药血清对脂多糖(LPS)诱导Caco-2细胞损伤模型的保护作用。方法收集对数生长期细胞,分为5个组,即正常对照组、LPS组,以及QCS低剂量组(2.5%含药血清)、中剂量组(5%含药血清)、高剂量组(10%含药血清)。采用MTT法检测细胞活力;ELISA法测定培养细胞上清液IL-8、IL-17和PGE2含量,以及MDA、GSH和SOD水平;划痕实验检测细胞体外迁移能力;Western blot技术检测RhoA、Rac1蛋白的表达。结果 QCS能拮抗LPS诱导的Caco-2细胞活性下降;与LPS组比较,QCS可显著减少LPS诱导的IL-8、IL-17和PGE2分泌(P<0.05),并减少LPS诱导的MDA释放,增加GSH和SOD的分泌(P<0.01);划痕实验48 h时QCS组的划痕宽度明显变小,细胞迁移能力增强(P<0.001),划痕的修复加快,且呈浓度依赖性改变;Western blot结果显示,QCS能有效逆转LPS所致RhoA、Rac1蛋白表达的抑制(P<0.05)。结论 QCS含药血清对LPS诱导的Caco-2细胞损伤有保护作用,可通过抑制炎症因子、抗氧化应激和促进结肠上皮细胞迁移来提高Caco-2的生存率。

Objective To investigate the protective effects of Qingchang Suppository(QCS)medicated serum on lipopolysaccharide(LPS)-induced Caco-2 cell injury model.Methods Caco-2 cells in logarithmic growth phase were collected and divided into five groups:normal control group,LPS group,low-dose QCS group(2.5%QCS medicated serum),medium-dose QCS group(5%QCS medicated serum)and high-dose QCS group(10%QCS medicated serum).MTT assay was used to detect cell viability;ELISA was used to determine the content of IL-8,IL-17 and PGE2,and the levels of MDA,GSH and SOD in the supernatant;scratch assay was employed to test cell migration ability in vitro;Western blot was used to detect the expression of RhoA and Rac1 proteins.Results QCS antagonized the decrease in Caco-2 cell viability induced by LPS.Compared with the LPS group,QCS significantly reduced the secretion of IL-8,IL-17 and PGE2 induced by LPS(P<0.05),lowered the release of MDA induced by LPS and increased the secretion of GSH and SOD(P<0.01);the scratch width in the QCS group decreased significantly and the cell migration ability increased at 48 h(P<0.001),which speeded up the repair of scratches in a concentration-dependent manner;Western blot results showed that QCS could effectively reverse LPS-induced inhibition of RhoA and Rac1 protein expression(P<0.05).Conclusion Qingchang Suppository medicated serum protects Caco-2 cells from LPS-induced injury and increases cell survival rate by inhibiting inflammatory factors and antioxidant stress and promoting migration of colonic epithelial cells.