摘要
目的探讨维生素D(vitamin D,VD)与铁联合对脂多糖(lipopolysaccharide,LPS)所致的结肠炎性细胞的影响及作用机制。方法将SW480细胞随机分为对照组(control),模型组(model,1μg/ml LPS),25(OH)D_(3)干预组[1μg/ml LPS+500 ng/ml 25(OH)D_(3)],铁干预组[1μg/ml LPS+1 mmol/L柠檬酸铁铵(ferric ammonium citrate,FAC)],联合干预组[1μg/ml LPS+500 ng/ml 25(OH)D_(3)+1 mmol/L FAC],干预24 h后收集细胞,免疫荧光检测各组细胞活性氧(reactive oxygen species,ROS)水平;酶联免疫吸附试剂盒(enzyme linked immunosorbent assay,ELISA)检测细胞IL-6、TNF-α、1,25(OH)2D3水平;western blots法检测各组细胞p38、p-p38、CYP27B1和CYP24A1蛋白表达水平。结果与对照组相比,模型组的ROS、IL-6和TNF-α水平明显增加(P<0.05),p38蛋白磷酸化水平也明显增加(P<0.05),CYP27B1蛋白表达水平增加,CYP24A1表达降低,25(OH)D_(3)+FAC联合干预组的1,25(OH)2D3水平增加(P<0.05);与模型组比较,25(OH)D_(3)+FAC联合干预组的ROS、IL-6和TNF-α水平明显降低(P<0.05),p38蛋白磷酸化水平与CYP27B1蛋白表达明显降低(P<0.05),CYP24A1表达增加。结论维生素D与铁联合能够有效降低LPS所致的结肠炎性细胞的炎症水平,该过程可能是由p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路介导的。
Objective To investigate the effect of vitamin D combined with iron on colonic inflammatory cells exposed to lipopolysaccharide(LPS)and the mechanism in volved.Methods SW480 cells were randomly divided into the control group,model group(1μg/ml LPS),25(OH)D_(3)intervention group[1μg/ml LPS+500 ng/ml 25(OH)D_(3)],iron intervention group(1μg/ml LPS+1 mmol/L FAC),and the combined intervention group[1μg/ml LPS+500 ng/ml 25(OH)D_(3)+1 mmol/L FAC].The cells were collected after 24 hours of intervention.The level of ROS production was detected by immunefluorescence.The levels of IL-6,TNF-αand 1,25(OH)2D3 were detected by ELISA kits.The protein expression levels of p38,p-p38,CYP27B1 and CYP24A1 were detected by Western blot.Results Compared with the control group,ROS,IL-6 and TNF-αlevels in the model group were significantly increased(P<0.05),so was the phosphorylation level of p38 protein(P<0.05).The expression of CYP27B1 protein was increased,and the CYP24A1 expression was decreased,The level of1,25(OH)2D3 in the combined intervention group was increased(P<0.05).Compared with the model group,levels of ROS,IL-6 and TNF-αin the combined intervention group were significantly decreased(P<0.05),the levels of p38 protein phosphorylation and CYP27B1 protein expression were significantly decreased(P<0.05),but the CYP24A1 expression was increased.Conclusion The combination of vitamin D and iron can effectively reduce the inflammatory level of colon inflammatory cells exposed to LPS,which may be mediated by p38 MAPK signaling pathway.
作者
邱服斌
谷思宇
赵艺敏
王煚楠
乔瑜
张静
QIU Fu-bin;GU Si-yu;ZHAO Yi-min;WANG Jiong-nan;QIAO Yu;ZHANG Jing(School of Public Health,Shanxi Medical University,Taiyuan 030001;Shanxi Provincial Health Commission Supervision and Inspection Center,Taiyuan 030000)
出处
《营养学报》
CAS
CSCD
北大核心
2022年第4期390-394,共5页
Acta Nutrimenta Sinica
基金
国家自然科学基金(No.81573156)
关键词
维生素D
铁
P38丝裂原活化蛋白激酶
活性氧
细胞培养
vitamin D
iron
p38 mitogen-activated protein kinase
reactive oxygen species
cell culture
作者简介
通信作者:邱服斌(1968-),男,博士,副教授,E-mail:fbqiu@126.com
