白屈菜碱对H2O2诱导的IPEC-J2细胞炎症损伤的预防作用
Preventive effect of chelidonine on H2O2-induced inflammatory injury of IPEC-J2 cells
林惠莹 1杨黎宇 1李家伟 1林福 2李健1
作者信息
- 1. 福建农林大学福建省兽医中药与动物保健重点实验室,福建福州 350002
- 2. 福建福德农业发展有限公司,福建福州 350000
- 折叠
摘要
集约化养殖模式造成猪肠道炎症损伤日趋严重,致使小肠上皮屏障功能受损,影响猪群健康度.为研究白屈菜碱(chelidonine)对猪小肠上皮细胞炎症损伤的保护作用,本试验使用H2O2构建猪小肠上皮细胞系(IPEC-J2 cells)炎症损伤模型,采用MTT法筛选适宜的白屈菜碱浓度,采用ELISA法检测炎症因子(IL-1 β、IL-6、IL-8、IL-10、TNF-α、TGF-β1)以及NO含量,采用qPCR法和 Western blot检测上述炎症因子及iNOS的mRNA和蛋白表达水平.结果发现,在2.5～100.0 mg/L质量浓度范围内,白屈菜碱对IP EC-J 2细胞无毒性;2.5、5.0、10.0 mg/L白屈菜碱可显著恢复H2O2诱导的IPEC-J2细胞存活率下降(P＜0.05).经H2O2造模后,NO含量极显著升高(P＜0.01)、IL-10含量显著降低(P＜0.05)、其余炎症因子含量显著升高(P＜0.05),iNOS及上述炎症因子的mRNA和蛋白表达水平均与ELISA检测结果趋势一致;而3个试验浓度的白屈菜碱均可显著地逆转以上变化.结果表明,白屈菜碱对H2O2诱导的IPEC-J2细胞炎症损伤具有显著的预防作用,具备开发成为防治猪肠炎药物的潜力.
Abstract
The intensive farming mode caused increasingly serious intestinal inflammatory injury and damaged intestinal epithelial barrier function,which affected the healthiness of swine.To in-vestigate the protective effect of chelidonine on inflammatory injury of porcine small intestinal epi-thelial cells,H2 O2 was used to construct the inflammatory damage model of IPEC-J2 cells,and the appropriate concentration of chelidonine was screened by MTT method,and the contents of inflam-matory factors(IL-1β,IL-6,IL-8,IL-10,TNF-α,TGF-β1)and NO were detected by ELISA meth-od,and the mRNA and protein expression levels of these inflammatory factors and iNOS were de-tected by qPCR and Western blot.The results showed that chelidonine had no toxicity to IPEC-J2 cells in the concentration range of 2.5-100.0 mg/L,and 2.5,5.0 and 10.0 mg/L chelidonine could significantly recover the H2O2-induced decline in the survival rate of IPEC-J2 cells(P＜0.05).Af-ter H2O2 modeling,the content of NO was significantly increased(P＜0.01),while the content of IL-10 was significantly decreased(P＜0.05),but the content of other inflammatory factors was significantly increased(P＜0.05).The mRNA and protein expression levels of iNOS and the above inflammatory factors were consistent with the ELISA results.And all the three concentrations of chelidonine could reverse the above changes.These results indicate that chelidonine has a signifi-cant preventive effect on H2O2-induced inflammatory damage of IPEC-J2 cells and has the poten-tial to be developed as a drug against enteritis of swine.
关键词
白屈菜碱/H2O2/炎症损伤/炎症因子Key words
chelidonine/H2O2/inflammatory injury/inflammatory factors引用本文复制引用
基金项目
福建省科技重大专项资助项目(2021NZ029023)
出版年
2024
NETL
NSTL
万方数据