Department of Anesthesiology, Lishui City People's Hospital, the Sixth Affiliated Hospital of Wenzhou Medical University, the First Affiliated Hospital of Lishui University, Zhejiang Province, Lishui, China.
Department of Gastroenterology, Lishui City People's Hospital, the Sixth Affiliated Hospital of Wenzhou Medical University, the First Affiliated Hospital of Lishui University, Zhejiang Province, Lishui, China.
Allergol Immunopathol (Madr). 2022 May 1;50(3):85-92. doi: 10.15586/aei.v50i3.569. eCollection 2022.
This study investigated the role of dexmedetomidine (DEX) in dextran sulfate sodium (DSS)-induced NCM460 cells.
The viability and apoptosis of NCM460 cells treated with DEX with or without DSS were detected by CCK-8 and terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) assay. The level of inflammatory factors and expression of inflammation-related proteins, tight junction proteins and Ras homolog gene family, member A/Rho-associated coiled-coil containing protein kinase (RhoA/ROCK) signaling-related proteins in NCM460 cells treated with DEX and/or U46619 (RhoA/ROCK agonist) and/or DSS were detected by the respective enzyme-linked immunosorbent assay (ELISA) kits and Western blot analysis. The permeability of NCM460 monolayers was examined with transepithelial electrical resistance (TEER) assay.
DEX had no effect on NCM460 cell viability. However, DEX improved the viability and barrier damage and suppressed the apoptosis and inflammation of DSS-induced NCM460 cells. Correspondingly, the expression of inflammation-related proteins was reduced and the expression of tight junction proteins was increased in DSS-induced NCM460 cells after treatment with DEX. In addition, RhoA/ROCK signaling was activated in NCM460 cells induced by DSS, which was suppressed by DEX. The protective effects of DEX on DSS-indued NCM460 cells were reversed by U46619.
DEX improved viability and barrier damage while suppressed apoptosis and inflammation in DSS-indued NCM460 cells by inhibiting RhoA/ROCK signaling pathway.
本研究旨在探讨右美托咪定（DEX）在葡聚糖硫酸钠（DSS）诱导的 NCM460 细胞中的作用。
通过 CCK-8 和末端脱氧核苷酸转移酶（TdT）dUTP 缺口末端标记（TUNEL）检测 DEX 联合或不联合 DSS 处理的 NCM460 细胞活力和凋亡情况。通过相应的酶联免疫吸附测定（ELISA）试剂盒和 Western blot 分析检测 DEX 和/或 U46619（RhoA/ROCK 激动剂）和/或 DSS 处理的 NCM460 细胞中炎症因子水平和炎症相关蛋白、紧密连接蛋白以及 Ras 同源基因家族成员 A/卷曲螺旋含卷曲螺旋蛋白激酶（RhoA/ROCK）信号相关蛋白的表达。通过跨上皮电阻（TEER）测定法检测 NCM460 单层的通透性。
DEX 对 NCM460 细胞活力无影响。然而，DEX 可改善 DSS 诱导的 NCM460 细胞活力和屏障损伤，抑制细胞凋亡和炎症。相应地，DEX 处理可降低 DSS 诱导的 NCM460 细胞中炎症相关蛋白的表达，增加紧密连接蛋白的表达。此外，DSS 诱导的 NCM460 细胞中 RhoA/ROCK 信号被激活，DEX 可抑制该信号。U46619 可逆转 DEX 对 DSS 诱导的 NCM460 细胞的保护作用。
DEX 通过抑制 RhoA/ROCK 信号通路改善 DSS 诱导的 NCM460 细胞活力和屏障损伤，同时抑制细胞凋亡和炎症。
