芪药鸡金粥含药血清通过TLR4/NF-κB通路对Caco-2炎症细胞模型的影响

Effect of Qi Yao Ji Jin Zhou Medicated Serum on the Expressions of Inflammatory Model of Caco-2 Cell through TLR4/NF-κB Pathway

目的探讨芪药鸡金粥含药血清是否通过TLR4/NF-κB信号通路影响人结肠癌上皮细胞(Caco-2)炎症模型增殖活力及细胞间紧密连接蛋白表达水平。方法Caco-2细胞分为空白组、模型组,高、中和低剂量含药血清组5组。采用四甲基氮唑盐比色法(MTT)检测,观察含药血清对Caco-2细胞炎症模型的增殖活力;运用酶联免疫吸附测定法(ELISA)检测Claudin-1、Occludin、JAM-1和ZO-1紧密连接蛋白的表达水平,及TLR4、NF-κBp50、NF-κBp65和磷酸化IκBα信号通路蛋白表达水平。结果与模型组比较,高、中、低含药血清组均可以促进细胞活力,以低剂量组活力最强,中剂量组次之,高剂量最弱,其中中剂量含药血清组和低剂量含药血清组与模型组比较,差异有统计学意义(P<0.05)。Caco-2细胞Claudin-1、Occludin、JAM-1和ZO-1蛋白表达水平以空白组最高,低、中、高剂量组次之,模型组最低,各组比较差异均有统计学意义(P<0.05)。Caco-2细胞TLR4、NF-κBp50、NF-κBp65和磷酸化IκBα蛋白表达水平,模型组最高,低、中、高剂量组次之,空白组表达水平最低,各组比较差异有统计学意义(P<0.05)。结论芪药鸡金粥含药血清可以促进Caco-2细胞炎症模型细胞增殖活力,修复Caco-2细胞炎症模型细胞间的紧密连接蛋白,其作用可能与抑制TLR4/NF-κB通路有关。

Objective To investigate Chinese medicated food Qi Yao Ji Jin Zhou medicated serum on the proliferation of human colon cancer epithelial cells(Caco-2)inflammation model and the expression of tight junction proteins through the TLR4/NF-κB signaling pathway.Methods Caco-2 cells were divided into 5 groups:blank group,model group,high-dose,middle-dose and low-dose drug-containing serum group.The tetramethylazole salt colorimetric method(MTT)was used to detect the proliferation activity of the drug-containing serum on the inflammation model of Caco-2 cells;the enzyme-linked immunosorbent assay(ELISA)was used to detect Claudin-1,Occludin,and JAM-1 And ZO-1 tight junction protein expression level,and TLR4,NF-κBp50,NF-κBp65 and phosphorylated IκBαsignaling pathway protein expression levels.Result Compared with the model group,high,medium and low levels of serum could promote Caco-2 cell proliferation.The expression levels of Claudin-1,Occludin,JAM-1 and ZO-1 claudins proteins were increased,and the expression levels of four proteins in the medium dose drug-containing serum group were significantly higher than those in the model group(P<0.05).ZO-1 protein was increased in the high,medium and low serum group(P<0.05).The expression levels of TLR4,NF-κBp50,NF-κBp65 and phosphorylated IκBαlevels are reduced.And the expression levels of NF-κBp50 protein in the low dose group and NF-κBp65 protein in the medium dose group were significantly reduced(P<0.05).Conclusion The Qi Yao Ji Jin Zhou medicated serum can promote the proliferation of Caco-2 cells and repair the tight junction protein between Caco-2 cells,which may be related to the inhibition of TLR4/NF-B pathway.