目的 探讨肠源性外泌体通过miR-146a调控NF-κB通路介导人肾小管上皮细胞HK-2的炎症反应。方法 Caco-2细胞分为对照组、模型［10 ng·mL^-1的脂多糖（LPS）刺激构建肠上皮细胞炎症模型］组、NC mimic组和miR-146a mimic组，提取各组外泌体（30 μg·mL^-1）孵育HK-2细胞，记为对照-Exos、LPS-Exos、NC-Exos、miR-146a-Exos组。转录组学测序对对照组与模型组差异基因进行检测；实时荧光定量PCR（qRT-PCR）检测miR-146a 表达；酶联免疫吸附 （ELISA） 法检测细胞培养基白细胞介素（IL）-1β、白细胞介素-6（IL-6）、肿瘤坏死因子α（TNF-α）水平；Western blotting法检测细胞中IL-1β、IL-6、TNF-α和核因子-κB（NF-κB）、磷酸化-NF-κB（p-NF-κB）、Toll样受体4（TLR4）炎性信号通路蛋白表达。结果 与对照组比较，模型组 Caco-2 细胞炎症因子 IL-1β、IL-6、TNF-α 和 TLR4、NF- κ B、p-NF- κ B 蛋 白 表 达 明 显增加（P＜0.05、0.01），培养基中 IL-1β、IL-6、TNF-α 水平显著增加（P＜0.05），细胞及外泌体中 miR-146a表达显著增加（P＜0.05、0.01）；miR-146a mimic组细胞及外泌体中miR-146a表达显著增加（P＜0.01）。外泌体与HK-2细胞共孵育后，与对照-Exos组比较，LPSExos、miR-146a mimic组培养基中炎症因子IL-1β、IL-6、TNF-α水平明显增加（P＜0.05），细胞中炎症因子IL-1β、IL-6、TNF-α及NF-κB炎症通路相关 TLR4、NF-κB、p-NF- κB 蛋白表达显著增加（P＜0.05、0.01）。结论 肠源性外泌体通过 miR-146a调控NF-κB通路介导HK-2细胞炎症反应。

Objective To investigate the role of intestine-derived exosomes in regulating the inflammatory response of HK-2 cells through the NF- κB pathway regulated by miR-146a. Methods Caco-2 cells were divided into control group, model group (constructed intestinal epithelial cell inflammation model by stimulating with 10 ng·mL^-1 LPS), NC mimic group, and miR-146a mimic group. The exosomes (30 μg·mL^-1) from each group were extracted and incubated with HK-2 cells, respectively, and named as control-Exos, LPS-Exos, NC-Exos, and miR-146a-Exos group. Transcriptome sequencing was used to detect differential genes between the control group and the model group; real-time fluorescence quantitative PCR (qRT-PCR) was used to detect miR-146a expression; ELISA was used to detect the levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) in the cell culture supernatant; Western blotting was used to detect the expressions of IL-1β, IL-6, TNF-α, NF-κB, p-NF-κB, and Toll-like receptor 4 (TLR4) inflammatory signaling pathway proteins in the cells. Results Compared with the control group, the expressions of inflammatory factors IL-1β, IL-6, and TNF- α and TLR4, NF- κB, and p-NF- κB proteins in the model group were significantly increased (P <0.05, 0.01), the levels of IL-1β, IL-6, and TNF-α in the cell culture supernatant were significantly increased (P <0.05), and the expressions of miR-146a in the cells and exosomes were significantly increased (P <0.05, 0.01); the expressions of miR-146a in the cells and exosomes of the miR-146a mimic group were significantly increased (P <0.01). Compared with the control-Exos group, the levels of inflammatory factors IL-1β, IL-6, and TNF-α in the culture supernatant of the LPS-Exos and miR- 146a mimic groups were significantly increased (P <0.05). Conclusion Intestine-derived exosomes regulate the inflammatory response of HK-2 cells through miR-146a-regulated NF-κb pathway.

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中山市社会公益与基础研究项目（2020B3005）；中山市第三批社会公益与基础研究项目（2021B3006）；天津市中医处课题（2021163）；天津市医药科学研究所课题（S202304）