肠激安方含药血清对脂多糖致炎人结肠上皮细胞活力、NF-κB/MLCK通路和紧密连接蛋白表达的影响OA

Aim:To investigate the effects of Changji'an Formula(CJA)on the viability of LPS-induced NCM460 cells and molecules involved in the NF-κB/MLCK pathway and the expression of tight junction proteins.Methods:NCM460 cells were divided into 4 groups.The normal control group was cultured in medium containing 10%(V/V)fetal bovine serum(FBS).The model group was cultured in medium containing 10%(V/V)FBS supplemented with 100 ng/mL lipopolysac-charide(LPS).The blank serum group was cultured in medium containing 10%(V/V)blank serum and 100 ng/mL LPS.The medicated serum group(CJA group)was cultured in medium containing 10%(V/V)CJA-serum and 100 ng/mL LPS.After 24 hours of incubation,cell viability was assessed using CCK-8 assay.The mRNA expression of NF-κB/MLCK path-way-related genes were determined by RT-qPCR,the expression of NF-κB/MLCK pathway-related proteins and tight junc-tion proteins were detected by Western blot.Results:Compared with the normal control group,the model group and blank serum group showed significantly reduced cell viability,decreased mRNA and protein expression levels of TRAF3,and re-duced expression of tight junction proteins;while the mRNA and protein expression levels of TNF-α and MLCK,as well as the expression levels of p-p65/p65 and p-MLC/MLC were significantly increased(P<0.05).Compared with the model group and blank serum group,CJA group showed significantly restored cell viability,increased TRAF3 mRNA and protein expression,reduced mRNA and protein expression levels of TNF-α,MLCK,p65 mRNA expression as well as the p65 mRNA expression level,as well as the expression levels of p-p65/p65 and p-MLC/MLC,and restored the expression levels of tight junction proteins(P<0.05).Conclusion:CJA can inhibit activation of NF-κB/MLCK pathway,improve the viability of NCM460 cells,and may contribute to the amelioration of membrane permeability.