BACKGROUND The specific and accurate pathogenesis of diarrhea-type irritable bowel syndrome is still unclear. AIMS We explored the mechanism of heat shock protein 27 (HSP27) in diarrhea-type irritable bowel syndrome to identify the key targets for the disease. METHODS The human colonic epithelial cell lines Caco-2 and NCM460 were pretreated with KRIBB3 (a phosphorylation inhibitor of HSP27) and then stimulated with lipopolysaccharide for different times. The apoptosis ratios of Caco-2 and NCM460 cells were examined with Annexin V/PI assays. Cell growth was determined using the cell counting kit-8 assay, and the expression levels of IL-1β and IL-6 in the cell supernatant were analyzed by ELISA. In addition, the expression levels of HSP27 and the nuclear factor-κB (NF-κB) signaling pathway were examined by Western blot assay. RESULTS Stimulation with lipopolysaccharide promoted the expression of HSP27 in colonic epithelial cells. HSP27 was phosphorylated at serine 78 and 82 after exposure to LPS. Apoptosis, growth inhibition, and inflammatory factor expression of lipopolysaccharide-induced colonic epithelial cells were greatly exacerbated by KRIBB3 treatment. In addition, KRIBB3 inhibited the phosphorylation of IκB-α and the activation of NF-κB. Gene silencing by small interfering RNA indicated that phosphorylation of HSP27 may regulate the NF-κB pathway. CONCLUSIONS HSP27 plays an important role in the inflammatory response of intestinal human colonic epithelial cells. HSP27 may protect intestinal epithelial cells against damage by regulating the NF-κB pathway.

中文翻译：

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热激蛋白27通过核因子-κB途径调节肠道上皮细胞的炎症反应。

背景技术腹泻型肠易激综合症的具体和准确的发病机制仍不清楚。目的我们探索了腹泻型肠易激综合征中热休克蛋白27（HSP27）的机制，以确定该疾病的关键靶标。方法用KRIBB3（HSP27的磷酸化抑制剂）预处理人结肠上皮细胞系Caco-2和NCM460，然后用脂多糖刺激不同时间。用膜联蛋白V / PI测定法检查了Caco-2和NCM460细胞的凋亡率。使用细胞计数试剂盒8测定法测定细胞生长，并通过ELISA分析细胞上清液中IL-1β和IL-6的表达水平。此外，通过蛋白质印迹法检测HSP27的表达水平和核因子-κB（NF-κB）信号通路。结果用脂多糖刺激可促进结肠上皮细胞中HSP27的表达。暴露于LPS后，HSP27在丝氨酸78和82处被磷酸化。KRIBB3处理可大大加剧脂多糖诱导的结肠上皮细胞的凋亡，生长抑制和炎性因子表达。此外，KRIBB3抑制了IκB-α的磷酸化和NF-κB的激活。小干扰RNA使基因沉默，表明HSP27的磷酸化可能调节NF-κB途径。结论HSP27在肠道人类结肠上皮细胞的炎症反应中起重要作用。HSP27可能通过调节NF-κB通路来保护肠上皮细胞免受损害。暴露于LPS后，HSP27在丝氨酸78和82处被磷酸化。KRIBB3处理可大大加剧脂多糖诱导的结肠上皮细胞的凋亡，生长抑制和炎症因子表达。此外，KRIBB3抑制了IκB-α的磷酸化和NF-κB的激活。小干扰RNA使基因沉默，表明HSP27的磷酸化可能调节NF-κB途径。结论HSP27在肠道人类结肠上皮细胞的炎症反应中起重要作用。HSP27可能通过调节NF-κB通路来保护肠上皮细胞免受损害。暴露于LPS后，HSP27在丝氨酸78和82处被磷酸化。KRIBB3处理可大大加剧脂多糖诱导的结肠上皮细胞的凋亡，生长抑制和炎性因子表达。此外，KRIBB3抑制了IκB-α的磷酸化和NF-κB的激活。小干扰RNA使基因沉默，表明HSP27的磷酸化可能调节NF-κB途径。结论HSP27在肠道人类结肠上皮细胞的炎症反应中起重要作用。HSP27可能通过调节NF-κB通路来保护肠上皮细胞免受损害。此外，KRIBB3抑制了IκB-α的磷酸化和NF-κB的激活。小干扰RNA使基因沉默，表明HSP27的磷酸化可能调节NF-κB途径。结论HSP27在肠道人类结肠上皮细胞的炎症反应中起重要作用。HSP27可能通过调节NF-κB通路来保护肠上皮细胞免受损害。此外，KRIBB3抑制了IκB-α的磷酸化和NF-κB的激活。小干扰RNA使基因沉默，表明HSP27的磷酸化可能调节NF-κB途径。结论HSP27在肠道人类结肠上皮细胞的炎症反应中起重要作用。HSP27可能通过调节NF-κB通路来保护肠上皮细胞免受损害。