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目的:探讨参苓白术散抗脂多糖(lipopolysaccharide,LPS)诱导肠上皮隐窝细胞(intestinal crypt cel ls,IEC-6)通透性变化的机制研究。方法:将IEC-6细胞分为空白组、LPS模型组、LPS+参苓白术散含药血清低、中、高剂量组、10%FBS。对以下指标进行检测:IEC-6细胞凋亡、细胞内钙离子浓度、IEC-6电阻值变化、检测各组中磷酸化Rho激酶(ROCKⅡ)、肌球蛋白轻链激酶(MLCK)以及凋亡因子Caspase-3表达。结果:LPS显著引起IEC-6细胞凋亡而参岺白术散的含药血清明显抑制细胞凋亡。与空白组比较,模型组IEC-6细胞跨膜电阻的TEER值显著下降(P<0.01);与LPS模型组比较,参苓白术散含药血清低、中、高剂量组IEC-6细胞跨膜电阻的TEER值显著升高,实验结果具有统计学意义(P<0.01)。LPS模型组的钙离子浓度明显高于空白组(P<0.01);不同浓度的参苓白术散含药血清组的钙离子浓度明显低于LPS模型组(P<0.01)。与空白组比较,模型组中磷酸化ROCKⅡ、MLCK蛋白表达明显升高(P<0.05);与模型组比较,不同剂量参苓白术散含药血清组中磷酸化ROCKⅡ、MLCK蛋白表达明显降低(P<0.05)。结论:参苓白术散含药血清对LPS诱导的IEC-6细胞损伤具有明显地抑制作用,与抑制细胞凋亡及IEC-6细胞通透性的改善有关。
Objective: To investigate the mechanism of Shenling Baizhu San on the changes of permeability of intestinal crypt cells (IEC-6) induced by lipopolysaccharide (LPS). Methods: IEC-6 cells were divided into blank group, LPS model group, LPS + Shenling Baizhu San containing low, medium and high dose serum, 10% FBS. The following indexes were tested: apoptosis of IEC-6 cells, intracellular calcium ion concentration and change of IEC-6 resistance value. The levels of ROCK II, MLCK, Factor Caspase-3 expression. Results: LPS markedly induced apoptosis in IEC-6 cells. Compared with the blank group, the TEER value of transmembrane resistance of IEC-6 cells in model group decreased significantly (P <0.01). Compared with LPS model group, IEC-6 cells in low, medium and high doses of Shenling Baizhu San containing serum The TEER value of membrane resistance increased significantly, the experimental results were statistically significant (P <0.01). The calcium concentration in LPS model group was significantly higher than that in blank group (P <0.01). The calcium concentration of Shenling Baizhu San containing serum at different concentrations was significantly lower than that in LPS model group (P <0.01). Compared with the blank group, the expression of phosphorylated ROCKⅡ and MLCK in the model group was significantly increased (P <0.05). Compared with the model group, the expressions of ROCKⅡ and MLCK in the SHR group were significantly decreased P <0.05). Conclusion: Shenling Baizhu San containing serum can significantly inhibit LPS-induced IEC-6 cell injury, which is related to the inhibition of apoptosis and the improvement of IEC-6 cell permeability.