Establishment of a lipopolysaccharide-induced inflammation model of human fetal colon cells

作者全名："Yu, Keqi; Liao, Shengtao; Li, Chuanfei; Song, Ya; Mei, Zhechuan; Lv, Lin"

作者地址："[Yu, Keqi; Liao, Shengtao; Li, Chuanfei; Song, Ya; Mei, Zhechuan; Lv, Lin] Chongqing Med Univ, Affiliated Hosp 2, Dept Gastroenterol, 74 Linjiang Rd, Chongqing 400010, Peoples R China"

通信作者："Mei, ZC; Lv, L (通讯作者)，Chongqing Med Univ, Affiliated Hosp 2, Dept Gastroenterol, 74 Linjiang Rd, Chongqing 400010, Peoples R China."

来源：MOLECULAR BIOLOGY REPORTS

ESI学科分类：MOLECULAR BIOLOGY & GENETICS

WOS号：WOS:000984633000009

JCR分区：Q3

影响因子：2.6

年份：2023

卷号：　

期号：　

开始页：　

结束页：　

文献类型：Article; Early Access

关键词：Human fetal colon; Lipopolysaccharide; Interleukin-6; Tumor necrosis factor-a

摘要："Background Inflammatory bowel disease (IBD) is a global health problem and there are few cell models for IBD at present. To culture a human fetal colon (FHC) cell line in vitro and establish an FHC cell inflammation model that meets the requirements for high expression of interleukin-6 (IL-6) and tumor necrosis factor-a (TNF-alpha). Methods and results FHC cells were cultured with various concentrations of Escherichia coli lipopolysaccharide (LPS) in appropriate media for 0.5, 1, 2, 4, 8, 16 and 24 h to stimulate an inflammatory reaction. The viability of FHC cells was detected by a Cell Counting Kit-8 (CCK-8) assay. The transcriptional levels and protein expression changes of IL-6 and TNF-a in FHC cells were detected by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and Enzyme-Linked Immunosorbent Assay (ELISA), respectively. Appropriate stimulation conditions were selected (i.e., LPS concentration and treatment time), based on changes in cell survival rate, and IL-6 and TNF-alpha expression levels. An LPS concentration higher than 100 mu g/mL or a treatment time longer than 24 h resulted in morphological changes and decreased cell survival. By contrast, expression levels of IL-6 and TNF-alpha significantly increased within 24 h when LPS concentration lower than 100 mu g/ mL and peaked at 2 h, whilst maintaining cell morphology and viability in FHC cells. Conclusion The treatment of FHC cells with 100 mu g/mL LPS within 24 h was optimal in terms of stimulating IL-6 and TNF-alpha expression."

基金机构：National Natural Science Foundation [82173360]; Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University [13-003-023]; Senior Medical Talents Program of Chongqing for Young and Middle-aged [11-020]; Chongqing medical scientific research project (Joint project of Chongqing Health Commission and Science and Technology Bureau) [2020GDRC014]; China Postdoctoral Science Foundation [2022M720606]; Special support for postdoctoral of Chongqing [2212013362080033]

基金资助正文："The present study was supported by the National Natural Science Foundation (grant no. 82173360), Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University (grant no. 13-003-023), Senior Medical Talents Program of Chongqing for Young and Middle-aged (grant no. 11-020), Chongqing medical scientific research project (Joint project of Chongqing Health Commission and Science and Technology Bureau) (grant no. 2020GDRC014), China Postdoctoral Science Foundation (grant no. 2022M720606), and Special support for postdoctoral of Chongqing (grant no. 2212013362080033)."