Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University, 74 Linjiang Road, Yuzhong, Chongqing, 400010, P.R. China.
Mol Biol Rep. 2023 Jul;50(7):5557-5564. doi: 10.1007/s11033-023-08465-7. Epub 2023 May 8.
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-α (TNF-α).
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-α 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-α expression levels. An LPS concentration higher than 100 µg/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-α significantly increased within 24 h when LPS concentration lower than 100 µg/mL and peaked at 2 h, whilst maintaining cell morphology and viability in FHC cells.
The treatment of FHC cells with 100 µg/mL LPS within 24 h was optimal in terms of stimulating IL-6 and TNF-α expression.
炎症性肠病（IBD）是一个全球性的健康问题，目前用于 IBD 的细胞模型很少。本研究旨在培养人胎儿结肠（FHC）细胞系，并建立一种符合高表达白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）要求的 FHC 细胞炎症模型。
用不同浓度的大肠杆菌脂多糖（LPS）在适当的培养基中培养 FHC 细胞 0.5、1、2、4、8、16 和 24 h 以刺激炎症反应。通过 Cell Counting Kit-8（CCK-8）测定 FHC 细胞的活力。通过定量实时聚合酶链反应（qRT-PCR）和酶联免疫吸附试验（ELISA）分别检测 FHC 细胞中 IL-6 和 TNF-α的转录水平和蛋白表达变化。根据细胞存活率和 IL-6 和 TNF-α表达水平的变化，选择适当的刺激条件（即 LPS 浓度和处理时间）。LPS 浓度高于 100 µg/mL 或处理时间超过 24 h 会导致形态变化和细胞存活率降低。相比之下，当 LPS 浓度低于 100 µg/mL 且在 2 h 时达到峰值时，在 24 h 内 FHC 细胞中 IL-6 和 TNF-α的表达水平显著增加，同时保持细胞形态和活力。
在 24 h 内用 100 µg/mL LPS 处理 FHC 细胞在刺激 IL-6 和 TNF-α表达方面是最佳的。
