摘要
目的 探讨蒲公英黄酮(DF)通过富含AT序列特异性结合蛋白2(SATB2)干预氧化应激和炎症反应对脂多糖(LPS)诱导的结肠上皮细胞损伤的保护作用.方法 对结肠上皮细胞FHC进行培养,将FHC细胞随机分为对照组(正常培养)、LPS组(10 μg·mL-1的LPS)、低剂量实验组(10 μg·mL-1的LPS+1 μmol·L-1 的 DF)、高剂量实验组(10 μg·mL-1 的 LPS+5 μmol·L-1 的DF)、高剂量+sh-NC 组(转染 sh-NC+10 μg·mL-1 的 LPS+5 μmol·L-1 的DF)、高剂量+sh-SATB2 组(转染sh-SATB2+10 μg·mL-1 的 LPS+5μmol·L-1的DF).用蛋白质印迹法检测各组FHC细胞SATB2蛋白相对表达水平;用四甲基偶氮唑蓝(MTT)检测各组FHC细胞存活率;用流式细胞术检测各组FHC细胞凋亡率;用试剂盒检测各组FHC细胞丙二醛(MDA)、白细胞介素-6(IL-6)水平.结果 对照组、LPS组、高剂量实验组、高剂量+sh-NC组、高剂量+sh-SATB2组细胞SATB2蛋白相对表达水平分别为0.83±0.09、0.19±0.03、0.66±0.05、0.62±0.07 和 0.23±0.03,细胞存活率分别为(100.00±1.00)％、(48.16±4.31)％、(85.31±5.83)％、(81.39±6.47)％和(58.75±5.24)％,细胞凋亡率分别为(3.27±0.81)％、(41.26±2.09)％、(11.35±1.04)％、(10.29±1.26)％和(35.87±2.15)％,MDA 含量分别为(13.16±1.73)、(52.87±3.49)、(23.19±2.05)、(20.98±3.17)和(44.87±3.05)μmol·L-1,IL-6 含量分别为(507.18±103.26)、(2 132.09±198.15)、(883.16±136.92)、(801.69±119.85)和(1 736.29±206.91)pg·mL-1.LPS组上述指标与对照组比较,在统计学上差异均有统计学意义(均P＜0.05);高剂量实验组上述指标与LPS组比较,在统计学上差异均有统计学意义(均P＜0.05);高剂量+sh-SATB2组上述指标与高剂量+sh-NC组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 DF通过SATB2干预氧化应激和炎症反应对LPS诱导的结肠上皮细胞损伤具有一定的保护作用.
Abstract
Objective To investigate the protective effect of dandelion flavone(DF)on lipopolysaccharide(LPS)-induced colon epithelial cell injury by intervening oxidative stress and inflammation with AT-specific binding protein 2(SATB2).Methods Colon epithelial cells FHC were cultured.FHC cells were randomly divided into control group(normal cultured),LPS group(10 μg·mL-1 LPS),experimental-L group(10 μg·mL-1 LPS+1 μmol·L-1 DF),experimental-H group(10 μg·mL-1 LPS+5 μmol·L-1 DF),experimental-H+sh-NC group(transfected with sh-NC+10 μg·mL-1 LPS+5 μmol·mL-1 DF),experimental-H+sh-SATB2 group(transfected with sh-SATB2+10 μg·mL-1 LPS+5μmol·L-1 DF).The relative expression level of SATB2 protein in FHC cells was detected by Western blotting.The survival rate of FHC cells in each group was determined by tetramethylazolium blue(MTT).The apoptosis rate of FHC cells in each group was detected by flow cytometry.The levels of malondialdehyde(MDA)and interleukin-6(IL-6)in FHC cells were detected by the kit.Results The relative expression levels of SATB2 protein in control group,LPS group,experimental-H group,experimental-H+sh-NC group and experimental-H+sh-SATB2 group were 0.83±0.09,0.19±0.03,0.66±0.05,0.62±0.07 and 0.23±0.03,respectively;cell viability rates were(100.00±1.00)％,(48.16±4.31)％,(85.31±5.83)％,(81.39±6.47)％and(58.75±5.24)％,respectively;cell apoptosis rates were(3.27±0.81)％,(41.26±2.09)％,(11.35±1.04)％,(10.29±1.26)％and(35.87±2.15)％,respectively;MDA levels were(13.16±1.73),(52.87±3.49),(23.19±2.05),(20.98±3.17)and(44.87±3.05)μmol·L-1,respectively;IL-6 levels were(507.18±103.26),(2 132.09±198.15),(883.16±136.92),(801.69±119.85)and(1 736.29±206.91)pg·mL-1,respectively.The above indicators in the LPS group showed significant differences compared to the control group(all P＜0.05);the above indicators in the experimental-H group showed significant differences compared to the LPS group(all P＜0.05);the above indicators in the experimental-H+sh-SATB2 group showed significant differences compared to the experimental-H+sh-NC group(all P＜0.05).Conclusion DF has a protective effect on LPS-induced colon epithelial cell injury by intervening oxidative stress and inflammation through SATB2.
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